Effect of Gegen Qinliantang on Fecal Short-chain Fatty Acids in Rats with Antibiotic-associated Diarrhea Based on Targeted Metabonomics / 中国实验方剂学杂志

ObjectiveTo explore the impact of Gegen Qinliantang（GQT） on the fecal short-chain fatty acids（SCFAs） metabolism in antibiotic-associated diarrhea（AAD） through targeted metabolomics. MethodA total of 240 SD rats were randomly divided into six groups（n=40， half male and half female）， including blank group， model group， bifidobiogen group（0.15 g·kg-1）， and GQT high-， medium-， and low-dose groups（10.08， 5.04， 2.52 g·kg-1）， except for the blank group， clindamycin（250 mg·kg-1） was given to all groups by gavage for modeling every day for 7 d. After successful modeling， each administered group was gavaged with the corresponding dose of the drug， and the blank and model groups were gavaged with an equal volume of normal saline solution， 1 time/d， for 14 d. At 0， 3， 7， 14 d after the drug intervention， eight rats were randomly selected from each group， respectively. Gas chromatography-time-of-flight mass spectrometry（GC-TOF-MS） was used to perform targeted metabolomic analysis of SCFAs in the feces of rats， and partial least squares-discriminant analysis（PLS-DA） was applied to compare the differences in metabolic profiles between groups at different treatment times， and to compare the changes in the contents of SCFAs in rat feces between groups. ResultPLS-DA results showed that the blank group could be clearly distinguishable from the model group， with GQT exhibiting a closer proximity to the blank group after 7 d of treatment. After further analyzing the composition of SCFAs， it was found that the proportion of acetic acid increased and the proportions of butyric acid， valeric acid， hexanoic acid and isovaleric acid decreased in the model group compared with the blank group. After the treatment with GQT， the proportions of butyric acid， isobutyric acid， valeric acid， and isovaleric acid increased， and the proportions of acetic acid， propionic acid and caproic acid decreased. Subsequent differential analysis revealed that GQT could significantly improve the content of butyric acid， and had a certain retrogressive effect on the contents of valeric acid and hexanoic acid. ConclusionThe medium dose group of GQT can improve the contents of SCFAs in AAD feces after 7 days of treatment， which may be related to the improvement of the composition ratio of SCFAs and the contents of butyric acid， valeric acid and caproic acid.

Exploring on Mechanism of Gegen Qinliantang in Interventing Antibiotic-associated Diarrhea Based on 16S rRNA Sequencing and Network Pharmacology / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Gegen Qinliantang（GQT） on the intestinal flora of antibiotic-associated diarrhea（AAD） by 16S rRNA sequencing and network pharmacology. MethodSixty SD rats were randomly divided into six groups（n=10）， including blank group， model group， GQT high-， medium- and low-dose groups（10.08， 5.04， 2.52 g·kg-1） as well as Lizhu Changle group（0.15 g·kg-1）， except for the blank group， each group was given clindamycin（250 mg·kg-1） by gavage once a day for 7 consecutive days. After successful modeling， the blank group and the model group were given equal volumes of normal saline by gavage. The other groups were given corresponding doses of drugs by gavage for 14 days. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） was used to screen the active components and targets of GQT， GeneCards， Online Mendelian Inheritance in Man（OMIM） database， Pharmacogenetics and Pharmacogenomics Knowledge Base（PharmGKB）， DrugBank and DisGeNET were used to search for AAD disease targets. The drug-disease common targets were obtained by R software. STRING was applied to analyze the target protein-protein interaction， and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis was performed. Then hematoxylin-eosin（HE） staining was used to observe the pathological changes of the colon， and 16S rRNA sequencing of AAD colon content flora structure further verified the results of network pharmacology. ResultThrough network pharmacology， it was found that 238 active components were screened from GQT and acted on 276 component targets， among which quercetin， puerarin， wogonin and apigenin were the main core components of GQT， 1 097 AAD disease targets and 127 drug-disease intersection targets. The protein-protein interaction network mainly included core targets such as protein kinase B1（Akt1）， interleukin（IL）-6 and IL-1β， which were mainly enriched in the IL-17 signaling pathway. It was verified through animal experiments that compared with the blank group， the colon structure of the model group was seriously abnormal， the intestinal epithelial columnar cells were damaged， the goblet cells were reduced， and a large number of inflammatory cells were infiltrated. Compared with the model group， the colon structure of the GQT high-dose group improved， but there were still abnormalities， the colon structure of GQT medium- and low- dose groups and Lizhu Changle group improved significantly and reached the normal level. GQT could improve the structural diversity of AAD intestinal flora. At the phylum level， the abundance of Firmicutes was increased and the abundance of Bacteroidetes was decreased. At the genus level， the abundance of Lactobacillus was increased， and the abundances of Prevotella and Bacteroides were decreased. Among them， Lactococcus could be used as a biomarker for AAD treatment with GQT， and the prediction of functional metabolism of intestinal flora revealed that GQT could promote acetate and lactate metabolic pathways in the intestine. ConclusionGQT may activate IL-17 signaling pathway by acting on the targets of Akt1 and IL-6 through key components such as quercetin and wogonin， and improve the abundance of Lactococcus in the intestinal tract as well as acetate and lactate metabolic pathways， so as to play a role in repairing the intestinal barrier for the treatment of AAD.

Effect of Bufeitang on Lung-gut Axis in Rats with Lung Qi-deficiency Syndrome of Chronic Obstructive Pulmonary Disease / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Bufeitang on intestinal flora of rats with lung Qi-deficiency syndrome of chronic obstructive pulmonary disease（COPD）， and to explore the mechanism of traditional Chinese medicine in regulating intestinal flora and thus restoring the balance of lung-gut axis. MethodA total of 84 rats were randomly divided into 7 groups， including blank group， model group， fecal bacterial transplantation（FMT） group， dexamethasone group and low， medium and high dose groups of Bufeitang， 12 rats in each group. Except for the blank group， cigarette and sawdust fumigation combined with intratracheal instillation of lipopolysaccharide（LPS） were used to establish the COPD rat model with lung Qi-deficiency syndrome in all other groups. The low， medium and high dose groups of Bufeitang were intragastric administrated with Bufeitang（3.645， 7.29， 14.58 g·kg-1）， the FMT group was given fecal bacteria liquid enema（10 mL·kg-1）， dexamethasone group was given dexamethasone acetate tablet suspension by gavage（0.135 mg·kg-1）， the blank group and model group were given equal amount of distilled water. Fresh feces were collected after 28 d of continuous intervention for 16S rRNA gene sequencing. Lung and colon tissues were stained with hematoxylin-eosin（HE） for pathomorphological observation， and enzyme-linked immunosorbent assay （ELISA） was performed to detect the contents of tumor necrosis factor-α（TNF-α） and interleukin-8（IL-8） in lung tissues. ResultCompared with the blank group， the model group showed severe abnormal lung tissue structure with alveolar atrophy and collapse accompanied by severe inflammatory cell infiltration. Compared with the model group， the extent of injury was significantly improved， and inflammatory cell infiltration was reduced with basically normal alveolar structure in the high dose group of Bufeitang. Compared with the blank group， the model group had severely abnormal colonic tissue structure， the epithelial cells in the mucosal layer were eroded and shed， the number of inflammatory cells increased， the submucosal layer was edematous and the gap was enlarged. Compared with the model group， the extent of damage was significantly improved in the medium and high dose groups of Bufeitang， the epithelial cells in the mucosal layer were neatly and closely arranged， with only a small amount of inflammatory cell infiltration and no significant degeneration. Compared with the blank group， the TNF-α and IL-8 levels of lung tissue in the model group were significantly increased（P<0.01）. Compared with the model group， the TNF-α and IL-8 levels of lung tissues in the low， medium and high dose groups of Bufeitang were significantly decreased（P<0.01）. Bufeitang significantly modulated the number of bacteria species as well as alpha and beta diversity of model rats， corrected the return of intestinal flora to normal abundance and diversity， and positively regulated 4 differential phyla（such as Firmicutes， Proteobacteria） and 13 differential genera（such as Turicibacter， Lactobacillus， Anaerobiospirillum， Intestinimonas） in COPD model rats with lung Qi-deficiency syndrome， and down-regulated 2 carbohydrate metabolic pathway functions， including the pentose phosphate pathway（non-oxidative branch） Ⅰ and the Calvin-Benson-Bassham cycle. ConclusionBufeitang can modulate the abundance and diversity of intestinal flora species， affect the function of metabolic pathways， repair the structure of lung and colon tissues， regulate the level of inflammatory factors， and thus improve COPD with lung Qi-deficiency syndrome. The mechanism may be related to its regulation of inflammation-related intestinal flora to restore the balance of lung-gut axis in COPD with lung Qi-deficiency syndrome.

Effects of changes in family structure on depressive anxiety and stress related symptoms among college students / 中国学校卫生

Objective@#To investigate the effects of major family structure changes on depression, anxiety and stress symptoms of college students, and to provide theoretical basis for mental health promotion and prevention.@*Methods@#A questionnaire survey was conducted among 9 779 college students from 6 universities, including Jiangxi University of Finance and Economics, Shangrao Normal University, Gannan Normal University, Fujian Polytechnic Normal University, and Changjiang University, by using the Depression, Anxiety and Stress Scale 21 Items (DASS 21).@*Results@#The prevalence rates of depression, anxiety and stress symptoms among college students were 27.4%, 42.0% and 17.4%, respectively. Univariate analysis showed that family structure was associated with anxiety and stress symptoms ( χ 2=8.40,13.08, P <0.05). Multivariate Logistic regression analysis showed that specific family structure other than single or two parent family was positively correlated with anxiety( OR =1.89,95% CI =1.05- 3.42 ) and stress symptoms ( OR =2.48, 95% CI =1.36-4.50), family structure changes due to parental divorce was positively correlated with stress symptoms ( OR =1.53,95% CI =1.05-2.20)( P <0.05).@*Conclusion@#The occurrence of depression, anxiety and stress symptoms of college students is related to the type of family structure and the changing factors. Colleges should pay more attention to the mental condition of college students with family structure changes, and deliver various mental health promotion services including psychological counseling and health education.

Long-term follow-up observation after vitrectomy in a family with vitreous amyloidosis due to transthyretin gene Gly83Arg mutation / 中华眼底病杂志

Objective:To investigate the causes of secondary glaucoma after vitrectomy for familial vitreous amyloidosis associated with transthyretin (TTR) gene Gly83Arg mutation.Methods:A retrospective case study. From January 2008 to January 2020, 13 cases (23 eyes) with hereditary vitreous amyloidosis and treated by vitrectomy in the Affiliated Hospital of Zunyi Medical University were collected. Among them, there were 7 males with 12 eyes and 6 females with 11 eyes. The average age was 43.0±4.8 years. All the affected eyes underwent standard three-channel vitrectomy through the flat part of the ciliary body. According to whether complete vitreous detachment (PVD) was formed during the operation, it was divided into complete PVD group and incomplete PVD group; according to the occurrence time of secondary glaucoma and vitreous amyloidosis after surgery, it was divided into 1-12 months group and 13-36 months group, >37 months group. The average follow-up time after surgery was 36.7±6.0 months. The incidence of secondary glaucoma and the recurrence rate of vitreous amyloidosis between groups were compared by χ2 test; the correlation between recurrence of vitreous amyloidosis and secondary glaucoma after surgery was analyzed by Spearman rank correlation analysis. Results:Among the 23 eyes, there were 8 eyes in the complete PVD group and 15 eyes in the incomplete PVD group, respectively. Vitreous amyloidosis recurred in 15 eyes (65.22%, 15/23) after surgery. There were 14 (93.30%, 14/15) and 1 (6.70%, 1/15) eyes in the incomplete PVD group and the complete PVD group, respectively; the comparison of the recurrence rate of vitreous amyloidosis between the two groups was statistically significant ( χ2=11.676, P<0.01). 1-12 months group, 13-36 months group, >37 months group included 1 (4.35%, 1/23), 12 (52.17%, 12/23), 2 (8.70%, 2/23) Only eye. The recurrence rate in the 13-36 months group was significantly higher than that in the 1-12 months group and >37 month group. Secondary glaucoma occurred in 11 eyes (47.80%, 11/23) after surgery. 1-12 months group, 13-36 months group, above 37 months group were 1 (4.35%, 1/23), 8 (34.78%, 8/23), 2 (8.70%, 2/23) eyes. The incidence of secondary glaucoma in the 13-36 months group was higher than that in the 1-12 months group and >37 months group. Among 11 eyes with secondary glaucoma, 10 eyes had recurrence of vitreous amyloidosis after surgery, and 1 eye had no recurrence. The results of Spearman rank correlation analysis showed that there was a positive correlation between the recurrence of vitreous amyloidosis and the occurrence of secondary glaucoma ( rs=0.516, P=0.012). Conclusion:The incidence of secondary glaucoma after vitrectomy in a family with vitreous amyloidosis caused by the Gly83Arg mutation of TTR gene is higher, and its occurrence is significantly positively correlated with the recurrence of vitreous amyloidosis.

Online and offline mixed teaching mode of medical genetics / 生物工程学报

Online courses are an indispensable part of medical teaching in the new era. Online courses have good prospects, although also with certain problems in practice. As an important basic medical course, medical genetics has both basic theoretical knowledge and clinical cases, involving basic principles and the latest developments. A single online course or offline teaching model cannot meet the needs of subject development and training a new generation of medical professionals. Therefore, actively exploring the online and offline hybrid teaching model is one of the important topics in the current medical teaching reform.

The influence of glucose regulatory protein 78 on prognosis and tumor cell proliferation of hepatocellular carcinoma / 中华消化外科杂志

Objective:To investigate the influence of glucose regulatory protein 78 (GRP78) on prognosis and tumor cell proliferation of hepatocellular carcinoma.Methods:The experimental study and retrospective cohort study were conducted. Based on hepatocellular carcinoma tissue chip, in vitro culture of Huh7 and Hep3B hepatoma cells and LO2 normal hepatic cell, and combined with immunohistochemical staining, cell transfection, quantitative real-time polymerase chain reaction (qRT-PCR), Western blot detection, cell proliferation experiments, cell clone formation experiments and high-throughput transcription histological analysis, the GRP78 expression in hepatoma cells was analyzed. Huh7 and Hep3B hepatoma cells being transfected with the GRP78 gene-specific shRNA lentiviruses or the negative control shRNA lentivirus were set as the GRP78 gene-specific shRNA lentivirus group and the negative control shRNA lentivirus group respectively. Observation indicators: (1) GRP78 expression in hepatocellular carcinoma tissue and adjacent tissue and its correlation with the clinicopathological characteristics of hepatocellular carcinoma patients; (2) analysis of factors affecting the prognosis of hepatocellular carcinoma patients; (3) effects of inhibiting of GRP78 expression on the proliferation of hepatoma cells; (4) effects of inhibiting of GRP78 expression on the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells; (5) effects of HA15 on the proliferation and the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells. Measurement data of the normal distribution were expressed as Mean± SD, and comparison of groups was conducted using the t test or ANOVA. Repeated measurement data were analyzed using repeated ANOVA. Count data were expressed as absolute numbers, and comparisons between groups was conducted using the chi-square test. COX proportional hazards regression model was used for univariate and multivariate analysis. The Kaplan-Meier method was used to calculate the survival time and draw survival curve, and the Log-rank test was used for generative analysis. Results:(1) GRP78 expression in hepatocellular carcinoma tissue and adjacent tissue and its correlation with the clinicopathological characteristics of hepatocellular carcinoma patients: results of immunohistochemical staining of hepatocellular carcinoma tissue chip showed that GRP78 was low-expressed in 53 cases and high-expressed in 37 cases of the 90 hepatocellular carcinoma tissues. GRP78 was low-expressed in 84 cases and high-expressed in 6 cases of the 90 paracancerous tissues. There was a significant difference in GRP78 expression between hepatocellular carcinoma tissues and paracancerous tissues ( P<0.05). (2) Analysis of factors affecting the prognosis of hepatocellular carcinoma patients: all 90 patients were followed up for 5 to 56 months, with a median follow-up time of 49 months. The median overall survival time and median disease progression-free survival time were 56 months and 53 months in the 53 hepatocellular carcinoma patients with GRP78 as low-expressed, versus 32 months and 19 months in the 37 hepatocellular carcinoma patients with GRP78 as high-expressed, respec-tively, showing significant differences ( χ2=17.482, 12.097, P<0.05). Results of univariate analysis showed that alanine aminotransferase (ALT), tumor pathological grading and GRP78 expression were related factors affecting the 3-year overall survival rate and disease progression-free survival rate of hepatocellular carcinoma patients ( hazard ratio=2.317, 2.039, 3.740 and 2.194, 2.177, 2.927, 95% confidence interval as 1.150?4.671, 1.201?3.462, 2.116?6.612 and 1.048?4.593, 1.093?4.336, 1.492?5.742, P<0.05). Results of multivariate analysis showed that ALT >40 U/L, tumor pathological grading as Ⅲ-Ⅳ grade and GRP78 as high-expressed were independent risk factors affecting the 3-year overall survival rate and disease progression-free survival rate of hepatocellular carcinoma patients ( hazard ratio=2.438, 2.245, 3.223 and 3.046, 2.473, 3.307, 95% confidence interval as 1.114?5.334, 1.047?4.814, 1.396?7.440 and 1.337?6.940, 1.141?5.360, 1.399?7.819, P<0.05). (3) Effects of inhibiting of GRP78 expression on the proliferation of hepatoma cells: ①results of qRT-PCR showed that the relative expression of GRP78 messenger RNA (mRNA) in Huh7, Hep3B, and LO2 cells were 3.06±0.33, 4.42±0.60 and 1.00±0.02. There were significant differences in GRP78 mRNA expression between Huh7 and LO2 cells or Hep3B and LO2 cells ( t=6.19, 5.42, P<0.05). ②Results of Western Blot detection showed that the relative expression of GRP78 protein in Huh7, Hep3B, and LO2 cells were 1.65±0.01, 1.77±0.01 and 0.99±0.02. There were significant differences in GRP78 protein expression between Huh7 and LO2 cells or Hep3B and LO2 cells ( t=75.09, 108.10, P<0.05). ③Results of cell proliferation experiments showed that the growth rates in Hu7 GRP78 gene-specific shRNA lentiviruses group cells and Hu7 negative control shRNA lentivirus group cells at 24, 48, 72 and 96 hours were 111.51%±0.35%, 144.85%±0.68%, 188.71%±3.62%, 282.51%±5.25% and 190.08%±0.58%, 285.76%±2.69%, 459.51%±4.29%, 597.88%±12.25%, showing signifi-cant differences ( Fgroups=1 360.000, Ftime=668.500, Finteraction=197.600, P<0.05). The growth rates in Hep3B GRP78 gene-specific shRNA lentiviruses group cells and Hep3B negative control shRNA lentivirus group cells at 24, 48, 72 and 96 hours were 124.47%±0.25%, 153.25%±1.25%, 195.45%±3.19%, 282.51%±10.76% and 179.69%±0.33%, 322.67%±2.46%, 486.27%±5.82%, 622.35%±12.58%, showing significant differences ( Fgroups=1 222.000, Ftime=706.200, Finteraction=179.600, P<0.05). ④Results of the cell clone formation experiments showed that the number of cells in Hu7 GRP78 gene-specific shRNA lentiviruses group cells and Hu7 negative control shRNA lentivirus group cells were 125±3 and 435±17, showing a significant difference ( t=17.86, P<0.05). The number of cells in Hep3B GRP78 gene-specific shRNA lentiviruses group cells and Hep3B negative control shRNA lentivirus group cells were 138±3 and 388±7, showing a significant difference ( t=32.29, P<0.05). (4) Effects of inhibiting of GRP78 expression on the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells: results of high-throughput transcription histological analysis showed that the relative expression rates of p53, p21, CDK2, CDK4, and CDK6 were 19%, 334%, 398%, 41% and 49% in the Hu7 GRP78 gene-specific shRNA lentiviruses group cells comparing to the Hu7 negative control shRNA lentivirus group cells. ①Results of qRT-PCR showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.17±0.03, 4.05±0.71, 3.73±0.47, 0.49±0.09, 0.48±0.06, 0.36±0.07 in the Hu7 GRP78 gene-specific shRNA lentiviruses group cells, versus 1.00±0.05, 1.03±0.17, 1.00±0.07, 1.01±0.09, 1.02±0.14, 1.00±0.03 in the Hu7 negative control shRNA lentivirus group cells, showing significant differences ( t=14.62, 4.17, 5.72, 4.26, 3.49, 8.82, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.11±0.01, 4.28±0.43, 4.19±0.22, 0.44±0.01, 0.25±0.03, 0.68±0.04 in Hep3B GRP78 gene-specific shRNA lentiviruses group cells, versus 1.01±0.09, 1.02±0.15, 1.00±0.06, 1.01±0.09, 1.01±0.08, 1.15±0.02 in Hep3B negative control shRNA lentivirus group cells, showing significant differences ( t=10.19, 7.14, 13.79, 6.37, 9.42, 9.61, P<0.05). ②Results of Western Blot detection showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.45±0.01, 1.98±0.05, 2.31±0.12, 0.75±0.03, 0.69±0.04, 0.82±0.03 in the Hu7 GRP78 gene-specific shRNA lentiviruses group cells, versus 1.01±0.05, 1.03±0.01, 1.00±0.02, 1.00±0.01, 1.01±0.02, 1.00±0.03 in the Hu7 negative control shRNA lentivirus group cells, showing significant differences ( t=11.07, 14.56, 11.30, 11.29, 10.55, 11.37, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.61±0.03, 1.98±0.16, 2.55±0.12, 0.85±0.03, 0.78±0.01, 0.54±0.02 in Hep3B GRP78 gene-specific shRNA lentiviruses group cells, versus 1.00±0.03, 1.05±0.02, 1.05±0.01, 1.05±0.02, 1.00±0.02, 1.00±0.02 in Hep3B negative control shRNA lentivirus group cells, showing significant differences ( t=10.97, 13.40, 12.35, 11.06, 12.45, 13.78, P<0.05). (5) Effects of HA15 on the proliferation and the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells: results of 50% inhibiting concentration (IC50) test of HA15 showed that the IC50 of HA15 for Huh7 and Hep3B cells at 48 hours were 9.98 μmol/L and 13.70 μmol/L. ①Huh7 and Hep3B cells were treated with 9.98 μmol/L and 13.70 μmol/L of HA15. Results of cell proliferation experiments showed that the growth rates at 24, 48, 72, and 96 hours were 112.81%±0.27%, 154.71%±1.45%, 237.66%±16.77%, 294.40%±14.92% in the HA15-Huh7 cells, versus 133.67%±0.49%, 352.93%±2.31%, 557.17%±4.89%, 662.60%±13.31% in the normal Huh7 cells, showing a significant difference ( Fgroups=766.800, Ftime=518.200, Finteraction=133.300, P<0.05). The growth rates at 24, 48, 72, and 96 hours were 121.27%±2.32%, 203.85%±3.18%, 240.80%±3.02%, 286.50%±7.10% in the HA15-Hep3B cells, versus 239.14%±1.02%, 362.00%±5.44%, 539.37%±10.80%, 694.79%±17.13% in the normal Hep3B cells, showing a signifi-cant difference ( Fgroups=594.300, Ftime=317.900, Finteraction=78.600, P<0.05). ②Results of qRT-PCR showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.27±0.05, 3.64±0.28, 4.13±0.41, 0.51±0.07, 0.39±0.03, 0.17±0.02 in the HA15-Huh7 cells, versus 1.02±0.14, 1.00±0.03, 1.00±0.05, 1.01±0.08, 1.01±0.09, 1.03±0.17 in the normal Huh7 cells, showing significant differences ( t=5.00, 9.25, 7.63, 4.73, 6.82, 5.01, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.28±0.03, 3.49±0.78, 4.31±0.53, 0.38±0.05, 0.36±0.04, 0.24±0.03 in the HA15-Hep3B cells, versus 1.01±0.11, 1.03±0.18, 1.01±0.08, 1.00±0.06, 1.02±0.15, 1.00±0.06 in the normal Hep3B cells, showing significant differences ( t=6.26, 3.08, 6.21, 7.97, 4.26, 11.08, P<0.05). ③Results of Western Blot detection showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.52±0.05, 1.94±0.08, 1.58±0.02, 0.89±0.00, 0.86±0.02, 0.74±0.01 in the HA15-Huh7 cells, versus 1.02±0.03, 1.00±0.03, 1.02±0.02, 1.04±0.03, 1.00±0.01, 1.01±0.02 in the normal Huh7 cells, showing significant differences ( t=11.54, 10.28, 11.03, 12.81, 13.67, 10.09, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.57±0.02, 1.67±0.04, 1.41±0.04, 0.82±0.03, 0.70±0.02, 0.74±0.01 in the HA15-Hep3B cells, versus 1.03±0.01, 0.98±0.03, 1.00±0.03, 1.03±0.03, 1.01±0.01, 1.04±0.01 in the normal Huh7 cells, showing significant differences ( t=10.81, 11.54, 12.26, 13.62, 14.23, 10.17, P<0.05). Conclusions:High expression of GRP78 is an independent risk factor affecting the overall survival and disease progression-free survival of hepatocellular carcinoma patients. Inhibiting of GRP78 expression can reduce cell proliferation and the expression of p53, p21, CDK2, CDK4, and CDK6 mRNA and proteins in hepatoma cells.

Isolation of Natural Fungal Pathogens from Marchantia polymorpha Reveals Antagonism between Salicylic Acid and Jasmonate during Liverwort-Fungus Interactions.

The evolution of adaptive interactions with beneficial, neutral and detrimental microbes was one of the key features enabling plant terrestrialization. Extensive studies have revealed conserved and unique molecular mechanisms underlying plant-microbe interactions across different plant species; however, most insights gleaned to date have been limited to seed plants. The liverwort Marchantia polymorpha, a descendant of early diverging land plants, is gaining in popularity as an advantageous model system to understand land plant evolution. However, studying evolutionary molecular plant-microbe interactions in this model is hampered by the small number of pathogens known to infect M. polymorpha. Here, we describe four pathogenic fungal strains, Irpex lacteus Marchantia-infectious (MI)1, Phaeophlebiopsis peniophoroides MI2, Bjerkandera adusta MI3 and B. adusta MI4, isolated from diseased M. polymorpha. We demonstrate that salicylic acid (SA) treatment of M. polymorpha promotes infection of the I. lacteus MI1 that is likely to adopt a necrotrophic lifestyle, while this effect is suppressed by co-treatment with the bioactive jasmonate in M. polymorpha, dinor-cis-12-oxo-phytodienoic acid (dn-OPDA), suggesting that antagonistic interactions between SA and oxylipin pathways during plant-fungus interactions are ancient and were established already in liverworts.

Evolution of an Eleusine-Specific Subgroup of Pyricularia oryzae Through a Gain of an Avirulence Gene.

Eleusine isolates (members of the Eleusine pathotype) of Pyricularia oryzae are divided into two subgroups, EC-I and EC-II, differentiated by molecular markers. A multilocus phylogenetic analysis revealed that these subgroups are very close to Eragrostis isolates. EC-II and Eragrostis isolates were exclusively virulent on finger millet and weeping lovegrass, respectively, while EC-I isolates were virulent on both. The avirulence of EC-II on weeping lovegrass was conditioned by an avirulence gene, PWL1. All EC-II isolates shared a peculiar structure (P structure) that was considered to be produced by an insertion (or translocation) of a DNA fragment carrying PWL1. On the other hand, all EC-I and Eragrostis isolates were noncarriers of PWL1 and shared a gene structure that should have predated the insertion of the PWL1-containing fragment. These results, together with phylogenetic analyses using whole-genome sequences, suggest that the Eleusine-specific subgroup (EC-II) evolved through a loss of pathogenicity on weeping lovegrass caused by a gain of PWL1.

Analysis on the Correlation Between ECG Parameters and Cardiac Function in Severe Triple Coronary Vessel Disease Patients Without Myocardial Infarction / 中国循环杂志

Objectives:To analyze the correlation between systolic cardiac insufficiency and ECG parameters of patients with triple-vessel disease(left anterior descending artery, left circumflex artery and right coronary artery showed ≥ 70% of diameter stenosis), but without history of myocardial infarction. Methods: A total of 96 triple-vessel disease patients without prior myocardial infarction who underwent coronary angiography examination between 2017-03-01 and 2017-07-05 in Zhongshan hospital were recruited in this study. According to LVEF, patients were divided into the normal cardiac function group (78 patients with LVEF ≥ 50%) and the reduced LVEF group (18 patients with LVEF < 50%). The Receiver Operating Characteristic (ROC) curve was applied to test optimal cut-off value of the ECG parameters and logistics regression analysis was utilized to determine the correlation between ECG indices with cardiac insufficiency. Results: A small percentage (18.8%) triple-vessel disease patients without prior myocardial infarction developed cardiac insufficiency. QRS duration, QTc duration were all significantly increased in patients with cardiac dysfunction) compared with patients with normal cardiac function (P < 0.05). ROC curve indicated good predictive efficacy to systolic cardiac insufficiency with HR > 70.5 bpm (sensitivity 81.3%, specificity 58.9%), QRS > 97.5 ms(sensitivity 82.4%, specificity 67.5%), QTc > 425 ms (sensitivity 93.8%, specificity 41.1%). Logistic multivariate regression analysis showed that QRS >97.5 ms(OR=7.577, 95%CI:1.094~52.490,P =0.030) was significantly correlated with systolic cardiac insufficiency. Conclusions: For triple-vessel disease patients without prior myocardial infarction, wider QRS in the resting ECG may indicate cardiac insufficiency.

Clinical analysis of 426 emmetropia patients with unilateral rhegmatogenous retinal detachment and retinal lesions in the contralateral eyes / 中华眼底病杂志

Objective To observe the fundus lesions in the unilateral rhegmatogenous retinal detachment (RRD) eyes and contralateral eyes in non-traumatic emmetropia patients,and explore the risks of lateral eyes.Methods This is a retrospective case analysis.A total of 426 patients of unilateral RRD diagnosed by clinical examination were enrolled in this study.There were 273 males and 74 females.The average age of onset was 54.7 years.81.46％ of them (347 patients) were 51-70 years old.The average detachment time was 2.12 months.They were divided into two groups,equal or lesser than 50 years old group and more than 50 years old group.A total of 100 patients (200 eyes) with ocular surface disorders were randomly selected as control.The lattice-like degeneration,cystic degeneration and dry retinal holes were treated with prophylactic laser photocoagulation.Follow-up period was 6 to 24 months.The age,gender,proliferative vitreous retinopathy (PVR) grading,best corrected visual acuity (BCVA),distribution and quantity of retinal holes,and posterior vitreous detachment (PVD) were retrospectively analyzed.The incidence of PVD among different age groups was compared with Chi square.Results Among 426 RRD eyes,there were 239 eyes (56.10％) with PVD.Among them,there were 30 eyes with age equal or lesser than 50 years old (12.55％) and 209 eyes with age more than 50 years old (84.75％).There were 187 eyes (43.90％) without PVD,which including 38 eyes with age equal or lesser than 50 years old (20.32％) and 149 eyes with age more than 50 years old (79.68％).The incidence of PVD among different age groups was statistically significant (x2=4.72,P＜ 0.05).There were 10,254,40 and 5 eyes in class A,B,C and D of PVR,respectively;117 eyes without PVR.The retinal hole was located in superior temporal,inferior temporal,superior nasal,inferior nasal and macular in 305,91,22,4 and 4 eyes,respectively.The number of holes was 1,2,and more than 3 in 297,89 and 40 eyes,respectively.The retinal detachment range of 1,2,3 quadrants and total dissociation were 92,230,71,33 eyes,respectively.The fundus lesion was found in 47 eyes (11.03％) in the lateral eyes.There were 20 RRD eyes in class B of PVR,and 27 RRD eyes in class C of PVR.Retinal degenerated area was found.Among them,the degeneration of 41 eyes was located in the temporal retina,45 eyes involved in a quadrant.There were 16 eyes with peripheral retinal dry holes;the holes diameter was less than 1,1-2,greater than 2 optic-discs in 6,11 and 5 retinal holes.At the end of the follow-up,there were 47 eyes with almost normal visual field,16 eyes with decreased visual acuity,noeyes with retinal detachment.In the control group,4 patients (5 eyes,2.50％) had fundus lesions.Conclusion The unilateral RRD in non-traumatic emmetropia mostly occurs in elderly patients;11.03％ of patients had fundus lesions in the contralateral eyes,higher than the general population.

The GYF domain protein PSIG1 dampens the induction of cell death during plant-pathogen interactions.

The induction of rapid cell death is an effective strategy for plants to restrict biotrophic and hemi-biotrophic pathogens at the infection site. However, activation of cell death comes at a high cost, as dead cells will no longer be available for defense responses nor general metabolic processes. In addition, necrotrophic pathogens that thrive on dead tissue, take advantage of cell death-triggering mechanisms. Mechanisms by which plants solve this conundrum remain described. Here, we identify PLANT SMY2-TYPE ILE-GYF DOMAIN-CONTAINING PROTEIN 1 (PSIG1) and show that PSIG1 helps to restrict cell death induction during pathogen infection. Inactivation of PSIG1 does not result in spontaneous lesions, and enhanced cell death in psig1 mutants is independent of salicylic acid (SA) biosynthesis or reactive oxygen species (ROS) production. Moreover, PSIG1 interacts with SMG7, which plays a role in nonsense-mediated RNA decay (NMD), and the smg7-4 mutant allele mimics the cell death phenotype of the psig1 mutants. Intriguingly, the psig1 mutants display enhanced susceptibility to the hemi-biotrophic bacterial pathogen. These findings point to the existence and importance of the SA- and ROS-independent cell death constraining mechanism as a part of the plant immune system.

Percutaneous Sacroiliac Screw Placement: A Prospective Randomized Comparison of Robot-assisted Navigation Procedures with a Conventional Technique / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Sacroiliac (SI) screw fixation is a demanding technique, with a high rate of screw malposition due to the complex pelvic anatomy. TiRobot™ is an orthopedic surgery robot which can be used for SI screw fixation. This study aimed to evaluate the accuracy of robot-assisted placement of SI screws compared with a freehand technique.</p><p><b>METHODS</b>Thirty patients requiring posterior pelvic ring stabilization were randomized to receive freehand or robot-assisted SI screw fixation, between January 2016 and June 2016 at Beijing Jishuitan Hospital. Forty-five screws were placed at levels S1 and S2. In both methods, the primary end point screw position was assessed and classified using postoperative computed tomography. Fisher's exact probability test was used to analyze the screws' positions. Secondary end points, such as duration of trajectory planning, surgical time after reduction of the pelvis, insertion time for guide wire, number of guide wire attempts, and radiation exposure without pelvic reduction, were also assessed.</p><p><b>RESULTS</b>Twenty-three screws were placed in the robot-assisted group and 22 screws in the freehand group; no postoperative complications or revisions were reported. The excellent and good rate of screw placement was 100% in the robot-assisted group and 95% in the freehand group. The P value (0.009) showed the same superiority in screw distribution. The fluoroscopy time after pelvic reduction in the robot-assisted group was significantly shorter than that in the freehand group (median [Q1, Q3]: 6.0 [6.0, 9.0] s vs. median [Q1, Q3]: 36.0 [21.5, 48.0] s; χ2 = 13.590, respectively, P < 0.001); no difference in operation time after reduction of the pelvis was noted (χ2 = 1.990, P = 0.158). Time for guide wire insertion was significantly shorter for the robot-assisted group than that for the freehand group (median [Q1, Q3]: 2.0 [2.0, 2.7] min vs. median [Q1, Q3]: 19.0 [15.5, 45.0] min; χ2 = 20.952, respectively, P < 0.001). The number of guide wire attempts in the robot-assisted group was significantly less than that in the freehand group (median [Q1, Q3]: 1.0 [1.0,1.0] time vs. median [Q1, Q3]: 7.0 [1.0, 9.0] times; χ2 = 15.771, respectively, P < 0.001). The instrumented SI levels did not differ between both groups (from S1 to S2, χ2 = 4.760, P = 0.093).</p><p><b>CONCLUSIONS</b>Accuracy of the robot-assisted technique was superior to that of the freehand technique. Robot-assisted navigation is safe for unstable posterior pelvic ring stabilization, especially in S1, but also in S2. SI screw insertion with robot-assisted navigation is clinically feasible.</p>

Acute necrotizing pneumonia combined with parapneumonic effusion caused by Mycobacterium lentiflavum: a case report.

BACKGROUND: Mycobacterium lentiflavum (M. lentiflavum), a slow growing nontuberculous mycobacterium (NTM), has recently been described as an emerging human pathogen regardless of the immune status of the host. Previous reports have demonstrated that cervical lymphadenitis of children is the most frequent pathology of M. lentiflavum. However, there are little reports regarding pulmonary diseases by M. lentiflavum specifically in immunocompetent patients. CASE PRESENTATION: A 60-year-old man having prolonged productive cough and dyspnea with fever was initially diagnosed as pneumonia with parapneumonic effusion. Imaging studies showed that the radiologic abnormality was acute bronchopneumonic infiltration with abscess formation in the left lower lobe and parapneumonic pleural effusion. M. lentiflavum was identified in the cultured pleural tissues. On the basis of these findings, he was diagnosed as pulmonary infection and pleurisy caused by M. lentiflavum, which was treated with a combination of antibiotics covering NTM. His clinical manifestations were dramatically improved by the treatment targeting NTM, while those were refractory to empirical antibiotic therapy. CONCLUSION: In this report, we introduce the isolation of M. lentiflavum from pleural tissues associated with acute necrotizing pneumonia combined with parapneumonic effusion in an immunocompetent host, suggesting that the M. lentiflavum can be a human pathogen invovled in pulmonary infectious diseases and pleurisy with poor response to empirical antibiotic treatment.

Repeated occurrence of second primary lung cancer at different sites in trachea: a case report.

Multiple or second primary lung cancers can develop at any sites in the lung with same or different histologic types, synchronously and/or metachronously. In case of metachronous occurrence of the second primary lung cancer, it is easy to confuse with the primary lung cancer as a recurrence of precedent lung malignancy treated successfully or metastasis. Previous reports have demonstrated that majority of the second primary lung malignancies have same histologic types regardless of their developing time and location. However, the repeated occurrence of the second primary lung malignancy, in particular with the different histologic features, is a very rare condition.A 62-year-old male who had past history of squamous cell carcinoma treated with surgery and adjuvant chemotherapy and the recurrence of lung malignancy on the trachea, which was also resected successfully visited our hospital due to blood tinged sputum. Evaluation using bronchoscopy and chest computed tomography revealed the tracheal mass looked similar grossly to the previous recurred tracheal mass that was resected surgically. Unexpectedly, the newly developed tracheal mass was confirmed as small cell lung cancer, the different histologic type from previous ones.In this report, we describe an interesting case of subsequent occurrence of second primary lung cancers showing histologic shifting at different sites in trachea, suggesting that it is important for physician to make an effort to identify the histologic characteristics of second primary lung cancers for the correct and adequate treatment no matter what they exhibit similar gross morphology.

Pulmonary chondroid hamartoma with nontuberculous mycobacterial infection: two case reports.

Solitary pulmonary nodules (SPNs) can be manifested in a variety of disorders including neoplasms, infection, inflammation, and vascular or congenital abnormalities. In addition, they are often accompanied with other pulmonary pathologic lesions such as consolidations and several pulmonary disorders present as similar pulmonary nodular lesions simultaneously. Diagnostic workup is important for these SPNs; however, many physicians often miss the second diagnosis for multiple pulmonary lesions with SPNs due to lack of clinical suspicion that each pulmonary nodule or pathologic lesion can have each other's diagnosis. Herein, we report 2 cases of coexistence of pulmonary chondroid hamartoma with nontuberculous mycobacterial (NTM) infection presenting as pulmonary nodules and multiple consolidative lesions. A 60-year-old man was admitted for the evaluation of multifocal pulmonary lesions including SPN with chronic exertional dyspnea. Multiple lung tissues were obtained from each lesion through percutaneous transthoracic needle biopsy (PTNB). At the same time, bacteriologic examination was performed using respiratory samples obtained by bronchoscopy. Based on pathologic and microbiologic results, the patient diagnosed as pulmonary chondroid hamartoma with pulmonary NTM infectious disease. In addition, a 56-year-old woman visited for the evaluation of a small SPN. The SPN was resected surgically for the pathologic examination and turned out to be pulmonary chondroid hamartoma. Interestingly, the diagnostic workup revealed that the patient had Lady Windermere syndrome which is one of features for Mycobacterium avium complex (MAC) pulmonary disease. Both patients were treated with the standard antibiotics against MAC as recommended by the ATS/IDSA guideline. This is the first report of 2 patients, as far as we know, that chondroid hamartoma and NTM disease develop simultaneously in the lung. This report emphasizes that physicians should endeavor to confirm the individual diagnosis for the various pulmonary abnormal lesions detected at the same time, if necessary through multifocal biopsies for each lesion.

Tonsillar metastasis of small cell lung cancer in a patient with idiopathic pulmonary fibrosis: a case report.

Small cell lung cancer (SCLC) metastasizes widely, but palatine tonsil is an extremely unusual site for metastasis. Idiopathic pulmonary fibrosis (IPF) is associated with increased risk of lung cancer. However, the most common histological findings among patients of lung cancer with IPF are known as non-SCLC such as adenocarcinoma and squamous cell carcinoma. In addition, the majority of them are located in IPF-associated fibrotic peripheral lesions. A 77-year-old man visited for 1-month persistent cough and dyspnea, with inspiratory dry crackles on both lower lung fields and a large oval mass in his throat. Chest computed tomography revealed 2 masses in the left lower lobe, 1 mass in the right upper lobe, and multiple enlarged mediastinal lymph nodes of the lung accompanying with IPF, which were diagnosed as SCLC pathologically. Very interestingly, the tonsillar mass was also confirmed as the metastatic lesion of SCLC. Chemotherapy for SCLC and medical treatment for IPF were applied. However, in following-up, he expired due to respiratory failure by an acute exacerbation of IPF 3 months after the diagnosis. In this current report, we describe, for the first time, a case of tonsillar metastasis of SCLC with IPF detected simultaneously in a 77-year-old man.

Blood-saving effect of prophylactic use of tranexamic acid in patients undergoing orthotopic liver transplantation / 中华麻醉学杂志

Objective To evaluate the blood-saving effect of prophylactic use of tranexamic acid in patients undergoing orthotopic liver transplantation.Methods Sixty ASA physical status Ⅰ-Ⅲ patients of both sexes,aged 18-60 yr,weighing 45-80 kg,scheduled for elective orthotopic liver transplantation,were randomly assigned to one of 2 groups (n =30 each) using a random number table:prophylactic use group (group P) and therapeutic use group (group T).Immediately after induction of anesthesia (T1),at 30 min of anhepatic phase (T2),and at 30 min and 2 h of neohepatic phase (T3,4),central venous blood samples were collected to determine plasma fibrinogen concentration (Fib) and platelet count,and the arterial blood samples were obtained to detect thromboelastography (TEG) parameters.In group T,when lysis after 30 min＞7.5％ and Clot Index ≤ 1.0 according to the results of TEG,which indicating that primary hyperfibrinolysis occurred,tranexamic acid 15-20 mng/kg was injected intravenously.In group P,immediately after beginning of skin incision,immediately after occlusion of portal vein,and immediately after portal vein unclamping,tranexamic acid 1 g was injected intravenously,and a single injection of tranexamic acid 15-20 mg/kg was given when primary hyperfibrinolysis occurred.The intraoperative blood loss,fluid input and output and transfusion of blood components were recorded.The duration of stay in ICU,amount of abdominal drainage during stay in ICU,volume of blood transfused within 72 h after operation,and hepatic artery and portal vein thrombosis within 1 week after operation were recorded.Results Compared with group T,the intraoperative blood loss,volume of succinylated gelatin injection transfused,and requirement for platelet and cryoprecipitate were significantly reduced,Angle at T2 and lysis after 30 min at T2,3 and maximum amplitude at T3 were increased,and no significant change was found in the duration of stay in ICU,postoperative amount of abdominal drainage and volume of blood transfused in group P.No patients developed primary hyperfibrinolysis in group P.No hepatic artery and portal vein thrombosis was detected within 1 week after operation in the two groups.Conclusion Prophylactic use of tranexamic acid can effectively prevent hyperfibrinolysis and reduce intraoperative blood loss without increasing the risk of development of thrombosis,and it provides better blood-saving effect than therapeutic use guided by TEG in patients undergoing orthotopic liver transplantation.

Mitochondrial pathway of retinal pigment epithelial cell apoptosis induced by blue light in vitro / 中华实验眼科杂志

Background Studies determined that blue light exposure causes apoptosis of human retinal pigment epithelial (RPE) cells,but its mechanism is still below understood.Objective The aim of this study was to investigate whether or how mitochondrial apoptotic pathway is involved in blue-light induced apoptosis of human RPE cells in vitro.Methods Human RPE cells were isolated from fresh donor eyes and primarily cultured and passaged.The cells were identified with keratin antibody by immunochemistry.Then the cells were the non-light exposed group,simple light-exposed group,light-exposed+nifedipine group,light-exposed+calphostin C group and the light-exposed+phorbol myristate acetate (PMA) group.Human RPE cells in light-exposed group were consequently cultured for 24 hours following the exposure of (2 000±500)lx blue-light for 6 hours,and then the expression levels of bax,bcl-2,bcl-xl in the cells were detected by Western blot to evaluate the effect of blue light on the apoptosis.The cells in the light-exposed+nifedipine group,light-exposed+calphostin C group and the light-exposed+PMA group were treated with the corresponding drugs 1 hour prior to light irradiation and sequently received 6-hour light irradiation and 48-hour culture.The expression of caspase-9 protein in the cells were assayed with Western blot to assess the influence of Ca2+ channel and protein kinase C (PKC) pathway on mitochondria of RPE cells.Results Cultured cells grew well with visible pigment in cytoplasm.The cells showed the positive response for keratin and presented a cobblestone-like appearance.The expression bands of bax,bcl-2 and bcl-xl proteins were clearly visible at the molecular weight of 23 000,26 000 and 30 000 in both non-light exposed group and the simple light-exposed group,and the absorbance values of the cells to bax were elevated,while the absorbance values to bcl-2 and bcl-xl were declined in the simple light-exposed group compared with the non-light exposed group (t =-4.409,P =0.012 ;t =7.575,P =0.002 ; t =6.068,P =0.004).Compared with the non-light exposed group,the absorbance values of caspase-9 were significantly raised in the simple light-exposed group,light-exposed+calphostin C group and the lightexposed+PMA group (P=0.005,0.002,0.000),but no significant difference between the non-light exposed group and light-exposed+nifedipine group (P=0.191).Compared with the simple light-exposed group,the expression level was considerably higher in the light-exposed + PMA group (P =0.005) ; while that in the light-exposed + nifedipine group or light-exposed+calphostin C group was not significantly different (P=0.057,0.643).Conclusions Blue light exposure induces apoptosis of RPE cells by up-regulating the expressions of bax and caspase-9 proteins and down-regulating the expressions of bcl-2 and bcl-xl.The mitochondrial apoptosis pathway and PKC pathway participate in blue-light induced apoptosis of human RPE cells in vitro.