[Effects of in vivo targeted carboxylesterase 1f gene knockdown on the Kupffer cells polarization activity in mice with acute liver failure].

Objective: To investigate the effect of targeted carboxylesterase 1f (Ces1f) gene knockdown on the polarization activity of Kupffer cells (KC) induced by lipopolysaccharide/D-galactosamine (LPS/D-GalN) in mice with acute liver failure. Methods: The complex siRNA-EndoPorter formed by combining the small RNA (siRNA) carrying the Ces1f-targeting interference sequence and the polypeptide transport carrier (Endoporter) was wrapped in ß-1, 3-D glucan shell to form complex particles (GeRPs). Thirty male C57BL/6 mice were randomly divided into a normal control group, a model group (LPS/D-GalN), a pretreatment group (GeRPs), a pretreatment model group (GeRPs+LPS/D-GalN), and an empty vector group (EndoPorter). Real-time fluorescent quantitative PCR and western blot were used to detect Ces1f mRNA and protein expression levels in the liver tissues of each mouse group. Real-time PCR was used to detect the expression levels of KC M1 polarization phenotypic differentiation cluster 86(CD86) mRNA and KC M2 polarization phenotypic differentiation cluster 163 (CD163) mRNA in each group. Immunofluorescence double staining technique was used to detect the expression of Ces1f protein and M1/M2 polarization phenotype CD86/CD163 protein in KC. Hematoxylin-eosin staining was used to observe the pathological damage to liver tissue. A one-way analysis of variance was used to compare the means among multiple groups, or an independent sample nonparametric rank sum test was used when the variances were uneven. Results: The relative expression levels of Ces1f mRNA/protein in liver tissue of the normal control group, model group, pretreatment group, and pretreatment model group were 1.00 ± 0.00, 0.80 ± 0.03/0.80 ± 0.14, 0.56 ± 0.08/0.52 ± 0.13, and 0.26 ± 0.05/0.29 ± 0.13, respectively, and the differences among the groups were statistically significant (F = 9.171/3.957, 20.740/9.315, 34.530/13.830, P < 0.01). The percentages of Ces1f-positive Kupffer cells in the normal control group, model group, pretreatment group, and pretreatment model group were 91.42%, ± 3.79%, 73.85% ± 7.03%, 48.70% ± 5.30%, and 25.68% ± 4.55%, respectively, and the differences between the groups were statistically significant (F = 6.333, 15.400, 23.700, P < 0.01). The relative expression levels of CD86 mRNA in the normal control group, model group, and pretreatment model group were 1.00 ± 0.00, 2.01 ± 0.04, and 4.17 ± 0.14, respectively, and the differences between the groups were statistically significant (F = 33.800, 106.500, P < 0.01). The relative expression levels of CD163 mRNA in the normal control group, the model group, and the pretreatment model group were 1.00 ± 0.00, 0.85 ± 0.01, and 0.65 ± 0.01, respectively, and the differences between the groups were statistically significant (F = 23.360, 55.350, P < 0.01). The percentages of (F4/80(+)CD86(+)) and (F4/80(+)CD163(+)) in the normal control group and model group and pretreatment model group were 10.67% ± 0.91% and 12.60% ± 1.67%, 20.02% ± 1.29% and 8.04% ± 0.76%, and 43.67% ± 2.71% and 5.43% ± 0.47%, respectively, and the differences among the groups were statistically significant (F = 11.130/8.379, 39.250/13.190, P < 0.01). The liver injury scores of the normal control group, the model group, and the pretreatment model group were 0.22 ± 0.08, 1.32 ± 0.36, and 2.17 ± 0.26, respectively, and the differences among the groups were statistically significant (F = 12.520 and 22.190, P < 0.01). Conclusion: Ces1f may be a hepatic inflammatory inhibitory molecule, and its inhibitory effect production may come from the molecule's maintenance of KC polarization phenotypic homeostasis.

[Visualization analysis of liver fibrosis diagnosis based on Citespace software].

Objective: To explore the research hotspots and development trends in the field of liver fibrosis diagnosis by using visualization methods. Methods: The relevant literatures on liver fibrosis diagnosis were downloaded from the China National Knowledge Infrastructure database. CiteSpace 5.3.R6 software was used to analyze the authors, institutions and key node information to explore the main research groups, institutions and research hotspots and development trends of this field. Results: The analysis showed that the main research authors were Cai Weimin, Guo Qiyong, etc. The main research institutions included were Shengjing Hospital affiliated to China Medical University, Shenzhen Third People's Hospital, etc. The current diagnosis of liver fibrosis was mainly focused on invasive (liver biopsy) and non-invasive (serology, imaging) diagnosis. The non-invasive diagnosis may be the research hotspot and direction of liver fibrosis in the future. Conclusion: The visualization analysis of liver fibrosis diagnosis by CiteSpace software can quickly and intuitively understand the basic knowledge and evolution of this field, as well as the main research directions, hot spots and future development trends.

[Early efficacy of percutaneous endoscopic cervical discectomy with multimodal neurophysiological monitoring].

Objective: To explore the clinical efficacy of percutaneous endoscopic cervical discectomy (PECD) assisted by neurophysiology monitoring (NM) in the treatment of cervical spondylotic radiculopathy (CSR). Methods: The clinical data of 55 patients with CSR treated in the Department of Spinal Surgery of Henan Provincial People's Hospital from April 2015 to May 2018 were analyzed retrospectively. Among them, 29 patients were treated with multi-mode NM-assisted PECD (NM group) and 26 patients with PECD alone (PECD group). The gender, age, operation time, bleeding volume, average hospital stay and complications between the two groups were recorded and compared. In addition, the visual analogue score (VAS) of neck and upper limb pain and the score of Japanese Orthopedic Association (JOA) were compared between the two groups before operation, 1 month after the operation and at the last follow-up. These data between groups were compared by independent sample t test. Results: All patients in both groups were followed-up for at least 18 months. Neck VAS and upper limb VAS scores of two groups at 1 month post operation (neck: 2.1±1.2, 2.0±1.1; upper lamb: 2.4±1.2, 2.2±0.8) and the last follow-up (neck:0.8±0.5, 0.7±0.5; upper lamb: 0.8±0.7, 0.8±0.5) decreased significantly when compared with those before the operation (neck: 6.0±1.0, 5.9±1.0; upper lamb: 7.1±0.9, 7.4±0.9) (t=12.670-27.305, all P<0.05). However, there was no significant difference between the two groups (t=-1.107-0.917, all P>0.05). JOA scores of two groups at 1 month after the operation (12.7±0.8, 12.6±0.8), and at the last follow-up (14.6±0.7, 14.4±0.8) were all improved significantly from those before the operation (11.1±1.0, 10.9±0.8) (t=-11.074, -14.829, -9.603, -13.086, all P<0.05); however, there was no significant difference between the two groups (t=0.842, 0.003, both P>0.05). There was also no significant difference in bleeding volume, and operation time between the two groups, (t=-0.615, -0.922, P>0.05) but the average hospital stay and incidence of complications in the NM group were significantly lower than those in the PECD group (t=-2.815, χ(2)=4.755, both P<0.05). Conclusion: Multimode NM-assisted PECD in the treatment of CSR achieves satisfactory results, reducing the average hospital stay, reducing complications and improving surgical safety.

Imaging features of pure mucinous breast carcinoma: correlation with extracellular mucus content.

AIM: To analyse the correlation between imaging features using multiple techniques and extracellular mucus content in pure mucinous breast carcinoma (PMBC). MATERIALS AND METHODS: A retrospective review of available images from 25 patients with 25 PMBC tumours was conducted, with ultrasonography (US), ultrasonic elastography (USE), mammography, and breast-specific gamma imaging (BSGI) available for 25, 15, 11, and eight patients, respectively. Microscopic slides from each tumour were evaluated for extracellular mucus content. The correlation between imaging features and mucus content was analysed using linear-by-linear association chi-square tests or Spearman's rank correlation analyses. RESULTS: On US images, a significant correlation was found between mucus content and echo pattern (p=0.042) and colour Doppler blood flow (p=0.032), with a trend that the lower mucus content present in tumours, the more likely they were detected with isoechoic echo and high blood flow. On USE images, a moderate negative correlation (r=-0.60, p=0.029) was observed between mucus content and tumour stiffness. On BSGI images, a strong negative correlation (r=-0.92, p=0.001) was shown between mucus content and lesion to non-lesion ratio (L/N) values of radioactivity counts. No significant correlation was found between mucus content and mammography imaging features (all p>0.05). CONCLUSION: Imaging features at US, USE, and BSGI correlated with extracellular mucus content in PMBC tumours, among which the L/N value using BSGI imaging is the most relevant feature.

[Efficacy and peripheral immunity analysis of allogeneic natural killer cells therapy in patients with hepatocellular carcinoma].

OBJECTIVE: To evaluate the safety and efficacy of allogeneic natural killer (NK) cells in the treatment of primary hepatocellular carcinoma (HCC), and to elucidate the mechanism of NK cells therapy. METHODS: Twenty-one patients with primary HCC treated with allogeneic NK cells at the Fifth Medical Center of the PLA General Hospital were followed up for 1 year. Peripheral blood mononuclear cells (PBMCs) were isolated from patient-related donors and cultured in vitro for 15 days and infused to the patients in two consecutive days. Clinical data and laboratory data were collected and analyzed, including survival, clinical features, imaging changes, hematology, immunology, and biochemical indicators to evaluate the safety and efficacy of allogeneic NK cell therapy. The changes of peripheral blood lymphocyte subsets after treatment were also analyzed to explore the possible anti-tumor mechanisms. RESULTS: (1) Of the 21 patients with primary HCC, 11 patients were treated once, 5 patients were treated twice, and 5 patients were treated 3 times. After allogeneic NK cells infusion, 10 patients had fever, 1 patient had slight hepatalgia and 1 patient had slight headache, no other adverse events occurred including acute and chronic graft-versus-host disease (GVHD). They resolved spontaneously within 8 hours without other treatment. (2) The total disease control rate was 76.2% during one-year follow-up. Among them, the patients with Barcelona clinic liver cancer (BCLC) stage A had a disease control rate of 100%, stable disease (SD) in 10 cases; BCLC stage B patients had a disease control rate of 60%, partial response (PR) in 1 case, and SD 2 in cases; BCLC stage C patients had a disease control rate of 50%, complete response (CR) in 1 case, and 2 cases of PR. (3) The frequencies of NK cells and CD8+ T cells in peripheral blood were significantly lower than that before at 24 hours after treatment, and the frequencies of CD4+ T cells and CD4/CD8 were significantly higher than the baseline. CONCLUSION: Allogeneic NK cells have good safety and efficacy in the treatment of primary HCC. The anti-tumor effect of the allogeneic NK cells may play an important role in the activation of the patient's natural immune system and delay disease progression, suggesting that allogeneic NK cells combined with sorafenib may be a very effective treatment for advanced HCC, and further large-sample multicenter randomized controlled clinical trials are needed to validate this result.

[Clinical characteristics of cardiomyopathies complicated with ventricular thrombosis].

Objective: To summarize the clinical characteristics of cardiomyopathy complicated with ventricular thrombosis. Methods: The clinical data of inpatients suffered from cardiomyopathy complicated with ventricular thrombosis in Fuwai Hospital between January 2015 and May 2019 were analyzed retrospectively. Results: A total of 125 cases were reviewed, and 24.8% were female. Dilated cardiomyopathy was the most common disease (62.4%), followed by arrhythmogenic right ventricular cardiomyopathy (ARVC) (13.6%) and hypertrophic cardiomyopathy (11.2%). There were 74.4% thrombosis in left ventricle, 12.8% in right ventricle and 12.8% in biventricle. The proportions of right ventricle thrombosis were higher in ARVC than in other cardiomyopathies (52.9% vs 6.5%, P<0.01). The majority suffered from cardiac function New York Heart Association (NYHA) Class Ⅲ (45.6%) and class Ⅳ (39.2%). The ratio of NYHA Class Ⅳ was higher in female patients than in male ones (25.8% vs 10.6%, P<0.05). In lab detection, positive results of D-Dimer and N terminal-pro B type natriuretic peptide (NT-proBNP) accounted for 72.8% and 97.6%, respectively. There were 2.5% patients died in the hospital or discharged because of the worsening of illness, the chances were higher in female than male patients (9.7% vs 0, P<0.01). Among these patients, one succumbed to massive ischemic stroke caused by ventricular thrombus detachment under standard anticoagulation therapy. Conclusions: Dilated cardiomyopathy is the most common cardiomyopathy complicated with ventricular thrombosis. The most common location of thrombosis is left ventricle. Right ventricle thrombosis is more common in ARVC. The majority suffer from moderate or severe cardiac dysfunction. Higer proportion of female patients suffer from anemia, severe condition and poor prognosis.

[The role of abnormal mitochondrial fusion and fission in PBDE-47-induced change in mitochondrial mass in PC12 cells].

Objective: To investigate the effect of 2, 2', 4, 4'-tetrabromodiphenyl ether (PBDE-47) on the mitochondrial mass in rat adrenal pheochromocytoma (PC12) cells and the potential mechanisms. Methods: Highly differentiated PC12 cells were divided into control, 1, 10 or 20 µmol/L PBDE-47-treated groups and cultured for 24 h. Transmission electron microscopy was employed to observe the changes in mitochondrial morphology and quantity in PC12 cells. Flow cytometry was used to measure the fluorescence intensity of Nonyl Acridine Orange (NAO) , a fluorescent indicator of mitochondrial membrane cardiolipin, to reflect mitochondria mass. Western blotting was used to determine the expression levels of Mitofusion 1 (Mfn1) and Fission 1 (Fis1) proteins. To further explore the role of abnormal mitochondrial fusion and fission in PBDE-47-induced mitochondrial mass changes, PC12 cells were divided into control group, 5 µmol/L M1 treatment group, 20 µmol/L PBDE-47 treatment group and 5 µmol/L M1+20 µmol/L PBDE-47 combined treatment group and cultured for 24 h, then the fluorescence intensity of NAO and expression levels of Mfn1 and Fis1 proteins were detected. Results: The control group showed numerous mitochondria with normal morphology, while the number of mitochondria decreased after PBDE-47 treatment. Especially, the disappeared cristae, swelling and vacuoles of mitochondria and decreased fluorescence intensity of NAO (P<0.05) were observed in 10 and 20 µmol/L PBDE-47-treated groups. Meanwhile, the expression levels of Mfn1 and Fis1 proteins in the 10 and 20 µmol/L PBDE-47-treated groups were significantly decreased compared with control group (P<0.05) . However, 5 µmol/L M1 co-treatment with 20 µmol/L PBDE-47 significantly increased the levels of Mfn1 and Fis1 proteins and fluorescence intensity of NAO compared with the 20 µmol/L PBDE-47 group (P<0.05) . Conclusion: PBDE-47 can inhibit the mitochondrial fusion and fission process, thus leading to damage of mitochondria mass in PC12 cells.

Negative pressure wound therapy inhibits inflammation and upregulates activating transcription factor-3 and downregulates nuclear factor-κB in diabetic patients with foot ulcerations.

BACKGROUND: Negative pressure wound therapy (NPWT) is one of the most important treatments for diabetic foot, but the underlying mechanisms of its benefits still remain elusive. This study aims to evaluate the inflammatory signals involved in the effects of negative pressure therapy on diabetic foot ulcers. METHODS: We enrolled 22 patients with diabetic foot ulceration, 11 treated with NPWT and the other 11 treated with traditional debridement. All patients were treated and observed for 1 week. Granulation tissues were harvested and analyzed in both groups, and then were histologically and immunohistochemically analyzed. Enzyme-linked immunosorbent assay, Western blot analysis, and real-time PCR were performed to evaluate the expression of interleukin-6 (IL-6), tumor necrosis factor α (TNF-α), inducible nitric oxide synthase (iNOS), nuclear factor-κB (NF-κB) p65, Ik B-α, and activating transcription factor-3 (ATF-3). RESULTS: After 7 days of treatment, NPWT could obviously promote diabetic wound healing because of the mild inflammation and the dense cell-deposited matrix. Meanwhile, NPWT significantly decreased the expression of TNF-α, IL-6, and iNOS (all P < .05). The result of Western blotting and real-time PCR indicated that NPWT obviously decreased the level of Ik B-α and NF-κB p65, and increased the level of ATF-3 (all P < .05). CONCLUSION: NPWT exerts an anti-inflammatory effect, possibly through the suppression of proinflammatory enzymes and cytokines resulting from Ik B-α inhibition and ATF-3 activation, which may prevent the activation of the NF-κB pathway in human diabetic foot wounds.

The association between GGCX, miR-133 genetic polymorphisms and warfarin stable dosage in Han Chinese patients with mechanical heart valve replacement.

WHAT IS KNOWN AND OBJECTIVE: Warfarin is a widely used anticoagulant with a narrow therapeutic index. Polymorphisms in the VKORC1, CYP2C9 and CYP4F2 genes have been verified to correlate with warfarin stable dosage (WSD). Whether any other genes or variants affect the dosage is unknown. The aim of our study was to investigate the relationship between GGCX, miR-133 variants and the WSD in Han Chinese patients with mechanical heart valve replacement (MHVR). METHODS: A total of 231 patients were enrolled in the study. Blood samples were collected for genotyping. The average WSD among subjects with different GGCX or miR-133 genotypes was compared. Regression analyses were performed to test for any association of genetic polymorphisms with WSD. RESULTS AND DISCUSSION: The warfarin dosage in patients with the GGCX rs699664 TT and rs12714145 TT genotypes was 3.77±0.93 (95% CI: 3.35-4.19) mg/d and 3.70±1.00 (95% CI: 3.32-4.09) mg/d, respectively. The GGCX rs699664 and rs12714145 genotypes were significantly associated with WSD (P<.05). But they were ruled out in the multivariate regression analysis. There were no significant differences in the average warfarin stable dosage between subjects with MIR133B rs142410335 wild-type and variant genotypes (P>.05). WHAT IS NEW AND CONCLUSION: The genotypes of GGCX rs699644 and rs12714145 were significantly associated with WSD (P<.05), but their contributions were not significant after accounting for other factors. MIR133B rs142410335 makes no significant contributions to warfarin stable dosage in Han Chinese patients with MHVR neither in univariate regression nor in multivariate regression analyses.

[Analysis of early failure of the PHILOS in proximal humerus fractures].

OBJECTIVE: To analyze the reasons of early failure of the PHILOS in proximal humerus fractures. METHODS: From Nov. 2010 to Nov. 2014, there were 117 patients with humerus fractures treated with PHILOS locking plate in Department of Orthopaedics, Xuanwu Hospital. All of the patients were treated with the plate by open reduction internal fixation, and we analyzed these cases retrospectively. After the operation, we removed the drainage tube within 48 h, and the patients were allowed to do the passive motion 3 days after the surgery if the X-Ray showed the plate and screws were reliable. Eight cases failed within 4 weeks after the operation. We analyzed the reasons of the failure. RESULTS: The rate of the failed cases was 6.83%(8/117). The average age was 72.4(66-82) years. In the 8 failed cases, 3 were on the right side, and the other 5 on the left side. As for the reason of the fractures, 2 cases were because of car accidents, and the other 6 because of daily life injury. According to the Neer classification, 3 cases were 2-part fractures, and the other 5 3-part fractures. Three cases were total failure, and the other 5 partial failure. All the 8 failed cases failed within 4 weeks after the operation, of which 1 was on the sixth day after surgery, the other 7 2 to 4 weeks after the surgery.The 3 totally failed cases were treated by removing the screws and plates, the other 5 by conservative methods. All of the cases were malunion at the end. CONCLUSION: The early failure of the PHILOS locking plate in proximal humerus fractures is related to the bad reduction during the operation, the loss of medial cortex support, the limitation of screw length, the osteoporosis and the improper rehabilitation after operation.It is very important to do good preoperative plan for a surgeon. During the operation, we should try our best in the fracture reduction, use the appropriate plate and screws, and then pay attention to the rehabilitation after the operation. After all of this, the rate of failure may be decreased.

[Analysis of early failure of the PHILOS in proximal humerus fractures].

OBJECTIVE: To analyze the reasons of early failure of the PHILOS in proximal humerus fractures. METHODS: From Nov. 2010 to Nov. 2014, there were 117 patients with humerus fractures treated with PHILOS locking plate in Department of Orthopaedics, Xuanwu Hospital. All of the patients were treated with the plate by open reduction internal fixation, and we analyzed these cases retrospectively. After the operation, we removed the drainage tube within 48 h, and the patients were allowed to do the passive motion 3 days after the surgery if the X-Ray showed the plate and screws were reliable. Eight cases failed within 4 weeks after the operation. We analyzed the reasons of the failure. RESULTS: The rate of the failed cases was 6.83%(8/117). The average age was 72.4(66-82) years. In the 8 failed cases, 3 were on the right side, and the other 5 on the left side. As for the reason of the fractures, 2 cases were because of car accidents, and the other 6 because of daily life injury. According to the Neer classification, 3 cases were 2-part fractures, and the other 5 3-part fractures. Three cases were total failure, and the other 5 partial failure. All the 8 failed cases failed within 4 weeks after the operation, of which 1 was on the sixth day after surgery, the other 7 2 to 4 weeks after the surgery.The 3 totally failed cases were treated by removing the screws and plates, the other 5 by conservative methods. All of the cases were malunion at the end. CONCLUSION: The early failure of the PHILOS locking plate in proximal humerus fractures is related to the bad reduction during the operation, the loss of medial cortex support, the limitation of screw length, the osteoporosis and the improper rehabilitation after operation.It is very important to do good preoperative plan for a surgeon. During the operation, we should try our best in the fracture reduction, use the appropriate plate and screws, and then pay attention to the rehabilitation after the operation. After all of this, the rate of failure may be decreased.

Relationship between the angle of vertebral screws and spinal lateral angulation after fixation of thoracolumbar fractures via an anterior approach.

This study investigated possible contributors to lateral spinal angulation after surgical fixation of thoracolumbar fractures via an anterior approach. We retrospectively examined lateral angulation in 172 cases of thoracolumbar fractures treated in this manner. The coronal Cobb angle and angles of the screws relative to the endplates were determined from radiographs. The patients completed the Short Form 36, Oswestry Disability Index, Japanese Orthopaedic Association Back Pain Evaluation Questionnaire, and Visual Analogue Scale at the final follow-up visit. The mean coronal Cobb angle was 0.75° ± 3.91° (-14.25° to 14.55°) preoperatively, 3.17° ± 4.07° (-8.18° to 14.01°) immediately postoperatively, and 3.46° ± 4.21° (-1.05° to 17.27°) at the final follow-up visit. The superior posterior and inferior anterior screws were more parallel to their respective endplates when the approach was made ≥2 vs ≤1 vertebral levels above the fracture (P < 0.001). Lateral angulation was more likely when the approach was made ≤1 vs ≥2 levels above the fracture (P < 0.001). The coronal Cobb angle differed significantly (P < 0.01) between patients with lumbar and thoracic fractures. The immediate postoperative coronal Cobb angle correlated tightly with the sum of the screw angles (superior plus inferior posterior and/or inferior plus superior anterior). Lateral angulation may occur after surgical fixation of thoracic and lumbar fractures via an anterior approach. Non-parallelism between the vertebral screws and their corresponding endplates may predict postoperative lateral spinal angulation. Postoperative lateral angulation does not correlate with low back pain, quality of life, or preoperative lateral angulation.

First Report of Leaf Blight of Rice Caused by Cochliobolus lunatus in China.

Leaf-streak symptoms were observed on rice (Oryza sativa L.) starting at the booting stage through harvest in Zhejiang Province, China, in 2012. Based on Fuyang County, only 15% of the rice fields were estimated to show these symptoms. However, incidence could be 40 to 80% when the rice got infected. Typical symptoms started as green water-soaked streaks from the tip or edge of leaf blades, similar to bacterial leaf blight caused by Xanthomonas oryzae. Infected leaves turned yellow, then eventually became wilted and dry. No bacterial streaming was observed and no bacteria were isolated. Pieces of infected leaf tissue were surface sterilized using 0.1% (v/v) mercuric chloride, rinsed with sterilized water, then placed on water agar (WA). After 2 or 3 days on WA at 28°C, only fungal growth was observed from surface sterilized tissues. Fungi were isolated, purified by single spore separation process, and subcultured to potato dextrose agar (PDA) plates. Growing on PDA, the surface of the colony was circular, fluffy, and shiny velvety-black, whereas the under surface was dark Prussian blue. Conidiophores were single or fascicled, brown to dark brown, rarely branched, multiseptate, and straight or often geniculate near the apex. Conidia were brown, smooth, fusiform, geniculate or hook-shaped, 17.5 to 28.5 × 8.5 to 14.0 µm, and 3-septate, with the third cell from the base larger and darker than the others. Molecular identification was performed by analysis of the rDNA internal transcribed spacer region (ITS1-5.8S-ITS2). The rDNA-ITS region was amplified with primer pair ITS1 and ITS4 (5), sequenced, and deposited in GenBank (Accession No. KC462186). The sequence of rDNA-ITS (KC462186) showed 100% identity with Cochliobolus lunatus R.R. Nelson & Haasis (JN943422) after BLAST. Based on the results of morphological and molecular analyses, the fungus isolated from infected leaves was identified as C. lunatus (anamorph: Curvularia lunata (Wakk.) Boedijn) (3). Pathogenicity tests were conducted three times by spraying a conidial suspension (1 × 105 spores/ml) with 0.1% (v/v) Tween 20 on 12 healthy rice plants at late tillering stage. The same number of the healthy rice plants sprayed with sterilized water with 0.1% (v/v) Tween 20 were used as control. All plants were kept at 30°C and 75 to 85% relative humidity (RH) under a 12-h light/dark rotation. About 5 to 7 days after inoculation, green water-soaked streaks began to appear on inoculated plants. From 7 to 14 days after inoculation, the lesions developed quickly and the leaves began to wilt. After 14 days, inoculated plants showed symptoms similar to those originally observed in the field, while control plants (sprayed with sterilized water) remained healthy. C. lunatus was re-isolated from all inoculated plants, and re-identified by the same methods (morphological and molecular methods) as described above, thereby satisfying Koch's postulates, and confirming C. lunatus as the cause of the disease. C. lunatus is a pathogen of a wide range of plants and is common in paddy environments. It was reported as one of the causal agents of black kernel of rice (4) and rice spikelet rot disease (SRD) (1,2). The level of incidence observed in the affected fields suggest that this disease could potentially cause major losses under favorable weather conditions if susceptible cultivars are grown. To our knowledge, this is the first report of C. lunatus causing leaf blight of rice in China. References: (1) S. W. Huang et al. Crop Prot. 30:1, 2011. (2) S. W. Huang et al. Crop Prot. 30:10, 2011. (3) D. S. Manamgoda et al. Fungal Divers. 51:3. (4) S. H. Ou. Rice diseases [M]. CABI, 1985. (5) T. J. White et al. PCR Protocols: a Guide to Methods and Application. Academic Press, San Diego, CA, 1990.

Development and comparison of a new personalized warfarin stable dose prediction algorithm in Chinese patients undergoing heart valve replacement.

BACKGROUND: Pharmacogenetics-based algorithms would be especially desirable for patients undergoing heart valve replacement (HVR), who are particularly sensitive to warfarin during the initial treatment phase following surgery. We aimed to derive a warfarin dosing algorithm from data of Chinese patients undergoing HVR, and to compare it with previously published dosing algorithms as applied to our HVR patients. METHODS: 641 Chinese HVR patients on stable maintenance dose of warfarin were enrolled from a single clinic site. Data of 321 patients were used to derive a warfarin dosing algorithm using stepwise multiple linear regression analysis. Previously published algorithms were selected from Pubmed database for comparison. The performance of all the algorithms was characterized according to mean absolute error (MAE) and percentage of predicted doses falling within +/- 20% of clinically observed doses (percentage of ideal prediction) in the other 320 patients. RESULTS: The newly developed algorithm included eight factors: VKORC1-1639G > A, CYP2C9*3, BSA, age, number of increasing INR drugs, smoking habit, preoperative stroke history and hypertension. Our algorithm accounted for 56.4% of variations in the inter-patient warfarin stable doses. All the algorithms showed better performance in a medium-dose (1.88-4.38 mg/day) and high-dose (> or = 4.38 mg/day) groupings than in a low-dose (< or = 1.88 mg/day) grouping. Compared with the 14 previously published algorithms, our algorithm had the lowest MAE (-0.07 mg/day) and the highest percentage of ideal prediction (62.8%) in the total validation cohort. CONCLUSIONS: Our warfarin dosing algorithm is potentially useful for patients whose population profiles are similar to those of our patients.

Characterizing the critical role of metabolism in osteosarcoma based on establishing novel molecular subtypes.

OBJECTIVE: Osteosarcoma is the third most frequently diagnosed cancer among adolescents. Immunotherapy is an effective curative treatment for metastatic osteosarcoma patients. This study aimed to further reveal the significance of metabolism in tumor progression, and to categorize molecular subtypes for guiding personalized therapy. MATERIALS AND METHODS: Univariate Cox regression analysis was performed to screen metabolism-related genes associated with osteosarcoma prognosis. A molecular subtyping system was developed by unsupervised consensus clustering. Survival analysis and functional analysis were used to evaluate the performance of subtyping and characterize the TME of subtypes. Stepwise Akaike information criterion (stepAIC) was employed to optimize the prognostic model. RESULTS: C1 and C2 subtypes showed distinct prognosis, with more favorable survival in C2 subtype. C2 subtype presented a higher immune infiltration and active anti-tumor response. Notably, C2 subtype was predicted to have better immune response to immune checkpoint blockade. In addition, a 5-gene prognostic signature with robust ability to classify patients into high-risk and low-risk groups was developed. CONCLUSIONS: The study revealed the critical role of metabolism in tumorigenesis by comparing the features between the two subtypes. Oncogenic pathways including epithelial mesenchymal transition (EMT), glycolysis and hypoxia may be closely involved in the correlation with metabolism. Importantly, we developed a novel subtyping system and a 5-gene signature with high potential to be applied in clinical practice.

[Immunogenicity and influencing factors of hepatitis B vaccination based on different vaccination schedules among chronic kidney disease patients].

Objective: To explore the immunogenicity and influencing factors of hepatitis B vaccination based on different vaccination schedules among chronic kidney disease (CKD) patients. Methods: CKD patients who participated in randomized controlled trials in four hospitals in Shanxi province and completed three doses of 20 µg vaccination (at months 0, 1 and 6) and four doses of 20 µg or 60 µg vaccination (at months 0, 1, 2, and 6) were surveyed from May 2019 to July 2020.According to the ratio of 1∶1∶1, 273 CKD patients were divided into 3 groups randomly. Quantification of the anti-hepatitis B surface antigen-antibody (anti-HBs) in serum samples was performed using chemiluminescent microparticle immunoassay at months 1 and 6 after the entire course of the vaccinations. The positive rate, high-level positive rate, geometric mean concentration (GMC) of anti-HBs, and the influencing factors were analyzed by χ2 tests, analysis of variance, unconditional logistic regression analysis. Results: A total of 273 CKD patitents were participants.The positive rates in the CKD patients with four doses of 20 µg vaccination (92.96%,66/71) or 60 µg vaccination (93.15%, 68/73) were higher than that in the CKD patients with three doses of 20 µg vaccination (81.69%, 58/71) at month one after the full course of the vaccinations (P<0.05). The GMCs of anti-HBs showed similar results (2 091.11 mIU/ml and 2 441.50 mIU/ml vs. 1 675.21 mIU/ml) (P<0.05). The positive rate was higher in the CKD patients with four doses of 60 µg vaccination (94.83%,55/58) than in those with three doses of 20 µg vaccination (78.79%,52/66) (P<0.05) at month six after the full course of the vaccinations. And the GMC of anti-HBs in the patients with four doses of 60 µg vaccination (824.28 mIU/ml) was significantly higher than those in the patients with 3 or 4 doses of 20 µg vaccination (639.74 mIU/ml and 755.53 mIU/ml) (P<0.05). After controlling the confounding factors, the positive rate in the CKD patients with four doses of 60 µg vaccination were 3.19 (95%CI: 1.02-9.96) and 5.32 (95%CI: 1.27-22.19) times higher than those in the patients with three doses of 20 µg vaccination at months 1 and 6 after the full course of the vaccinations, respectively. The positive rate in CKD patients without immune suppression or hormone therapy was 3.33 (95%CI: 1.26-8.80) and 4.78 (95%CI: 1.47-15.57) times higher than those in the patients with such therapy, respectively. Conclusions: Four doses of 20 µg or 60 µg hepatitis B vaccination could improve the immunogenicity in patients with CKD. And four doses of 60 µg vaccination might play a positive role in maintaining anti-HBs in this population. The immunogenicity in the CKD patients with immune suppression or hormone therapy was poor.

Protective effect of Gö6976, a PKD inhibitor, on LPS/D: -GalN-induced acute liver injury in mice.

OBJECTIVE: Protein kinase D (PKD) is a newly described serine/threonine protein kinase that plays a pivotal role in inflammatory response. In the present study, we examined the protective effect of Gö6976, a PKD inhibitor, on lipopolysaccharide (LPS) and D: -galactosamine (D: -GalN)-induced acute liver injury in mice. MATERIALS AND METHODS: Mice were pretreated intraperitoneally with Gö6976 30 min before LPS/D: -GalN administration . The mortality and degree of hepatic injury was subsequently assessed. RESULTS: The results indicated that LPS/D: -GalN administration markedly induced hepatic PKD activation, lethality and liver injury, while pretreatment of the PKD inhibitor Gö6976 significantly inhibited LPS-induced PKD activation, improved the survival of LPS/D: -GalN-administered mice and attenuated LPS/D: -GalN-induced liver injury, as evidenced by reduced levels of serum aminotransferases as well as reduced histopathological changes. In addition, the protective effects of Gö6976 were paralleled by suppressed activation of mitogen-activated protein kinases (MAPKs), decreased expression of tumor necrosis factor-α (TNF-α) and adhesion molecules, and reduced apoptosis and myeloperoxidase (MPO) activity in liver. CONCLUSIONS: Our experimental data indicated that Gö6976, a PKD inhibitor, could effectively prevent LPS/D: -GalN-induced acute liver injury by inhibition of MAPKs activation to reduce TNF-α production. This suggests the potential pharmacological value of PKD inhibitors in the intervention of inflammation-based liver diseases.

Antigen acquisition by dendritic cells: intestinal dendritic cells acquire antigen administered orally and can prime naive T cells in vivo.

In the rat, mesenteric lymphadenectomy allows collection of dendritic cells (DC) derived from the small intestine after cannulation of the thoracic duct. We prepared rats this way and administered antigens by oral feeding or intraintestinal injection. DC enriched from the thoracic duct lymph collected over the first 24 h from these animals are able to stimulate sensitized T cells in vitro and to prime popliteal lymph node CD4+ T cells after footpad injection, while B and T cells from the same thoracic duct lymph are inert in priming. 500 or less DC pulsed in vitro with antigen can prime T cells in vivo, whereas 100 times more B cells or macrophages pulsed in vitro are quite inert. 1 mg of ovalbumin administered orally is sufficient to load DC for in vivo priming of T cells. Antigen could not be detected directly in DC but was present in macrophages in the lamina propria. Direct presentation of antigen by DC to T cells was demonstrated by injecting F1 recipients with parental DC and showing restriction of T cell sensitization to the major histocompatibility complex of the injected DC. Antigen-bearing DC do not induce a detectable primary antibody response but a small secondary antibody response can be detected after a boosting injection. These results show that acquisition of antigens by DC in the intestine is very similar to what occurs in vitro or in other tissues, suggesting that there may be no special difference in antigen handling at mucosal surfaces. One implication of these results is that hypotheses designed to explain oral tolerance must take into account the presence of immunostimulatory, antigen-bearing DC in animals that have received oral antigens.

Pim-3 protects against hepatic failure in D-galactosamine (D-GalN)-sensitized rats.

BACKGROUND: Fulminant hepatic failure (FHF) has a high mortality resulted from massive hepatic apoptosis and haemorrhage necrosis; it is required to develop a valid therapy directed towards hepatocyte protection and regeneration. Pim-3, a hepatic growth stimulator, belongs to the serine/threonine kinase Pim-family that has been implicated in gp130-mediated induction of cell proliferation, protection from apoptosis downstream of Signal transducer and activator of transcription 3 (STAT3) and vascular endothelial growth factor-A-dependent vasculogenesis and angiogenesis, thus is suggested to possibly play a role in the tissue repair of FHF. MATERIALS AND METHODS: Male Wistar rats received simultaneous intraperitoneal injections of lipopolysaccharide (LPS) (100 microg kg(-1)) and D-galactosamine (D-GalN) (600 mg kg(-1)). One day prior to LPS/D-GalN administration, naked plasmid or Ringer's solution was injected via tail vein by hydrodynamics-based procedure. RESULTS: Exogenous Pim-3 gene protected against LPS/D-GalN-induced lethality with survival rate of more than 80% and improved the hepatic pathomorphism. The fractions of hepatic apoptotic-positive cells and the levels of caspase-3 activity were markedly lower in Pim-3-pretreated rats. Furthermore, exogenous Pim-3 significantly inhibited expression of tumour necrosis factor-alpha and interleukin-1beta in the liver, declined p53 and inducible nitric oxide synthase mRNAs levels, but elevated levels of Bcl-2 protein, an anti-apoptosis member of Bcl-2 family, in the liver. Exogenous Pim-3, however, showed little effect on expression of Bax, a pro-apoptosis member of Bcl-2 family. CONCLUSIONS: Pim-3 gene could protect rats from FHF by inhibiting liver apoptosis and improving inflammatory response of liver tissues, which could be associated with inhibiting expression of inflammatory mediators and promoting expression of anti-apoptosis protein Bcl-2.