[Early effects of unilateral biportal endoscopic posterior total laminectomy decompression in the treatment of cervical stenosis].

To investigate the early efficacy and safety of unilateral biportal endoscopic posterior total laminectomy decompression in the treatment of cervical stenosis (CS). The clinical data of 28 patients with CS treated by unilateral biportal endoscopic posterior total laminectomy decompression from January 2021 to October 2022 in the Henan NO.3 Provincial People's Hospital were retrospectively analyzed. Of the patients, 16 were male and 12 were female, the mean age was (55.6±9.6) years. The CS occurred at C3-4 level in 1 cases, at C4-5 level in 3 cases, at C5-6 level in 16 cases and at C6-7 level in 8 cases. Each case was compared at the moment of pre-operation and final follow-up by the Japanese Orthopedic Association (JOA) score. The postoperative complications were recorded. The JOA improvement rate was computed at the final follow-up. As a result, all patients underwent successful surgery and were followed up for (11.6±4.7) months. The operation time was (43.0±5.3) min. Intraoperative blood loss (7.9±2.8) ml; Postoperative drainage volume (8.1±2.3) ml. The JOA score increased from 7.9±1.2 before surgery to 13.5±1.3 six months after surgery, and it was 13.7±1.2 at the last follow-up, the differences between postoperative and preoperative were both statistically significant (both P<0.001). No complications occurred, such as cerebrospinal fluid leakage, nerve injury and intraspinal hematoma. At the last follow-up, cervical spine X-ray or CT evaluation showed no instability in the operative segment. The overall curative effect was evaluated according to JOA score at the last follow-up: 16 cases got excellent outcome, 7 cases got good and 2 cases got medium outcome, with an excellent and good rate of 89.3% (25/28). This study shows that unilateral biportal endoscopic posterior total laminectomy decompression in the treatment of single-level cervical stenosis can achieve satisfactory efficacy, has no impact on spinal stability, and has a high safety.

Host response of Nicotiana benthamiana to the parasitism of five populations of root-lesion nematode, Pratylenchus coffeae, from China.

In a recent survey of nematodes associated with tobacco in Shandong, China, the root-lesion nematode Pratylenchus coffeae was identified using a combination of morphology and molecular techniques. This nematode species is a serious parasite that damages a variety of plant species. The model plant benthi, Nicotiana benthamiana, is frequently used to study plant-disease interactions. However, it is not known whether this plant species is a host of P. coffeae. The objectives of this study were to evaluate the parasitism and pathogenicity of five populations of the root-lesion nematode P. coffeae on N. benthamiana.N. benthamiana seedlings with the same growth status were chosen and inoculated with 1,000 nematodes per pot. At 60 days after inoculation, the reproductive factors (Rf = final population densities (Pf)/initial population densities (Pi)) for P. coffeae in the rhizosphere of N. benthamiana were all more than 1, suggesting that N. benthamiana was a good host plant for P. coffeae.Nicotiana. benthamiana infected by P. coffeae showed weak growth, decreased tillering, high root reduction, and noticeable brown spots on the roots. Thus, we determined that the model plant N. benthamiana can be used to study plant-P. coffeae interactions.

[Early efficacy and safety of mini open (air/water medium) endoscopy assisted anterior cervical discectomy and fusion in the treatment of cervical spondylotic myelopathy].

Objective: To investigate the efficacy and safety of mini open (air/water medium) endoscopy assisted anterior cervical discectomy and fusion (MOEA-ACDF) for the treatment of cervical spondylotic myelopathy (CSM). Methods: A follow-up study. The clinical data of 30 patients with CSM treated by MOEA-ACDF from January to December in 2021 in the Henan NO.3 Provincial People's Hospital were retrospectively analyzed. Of the patients, 20 were male and 10 were female, the mean age was (49.8±9.3) years (ranged 28-70 years). The CSM occurred at C3-4 level in 2 cases, at C4-5 level in 3 cases, at C5-6 level in 22 cases and at C6-7 level in 3 cases. Each case was compared at the moment of pre-operation and final follow-up by the Japanese Orthopedic Association (JOA) score, C2-7 Cobb angle, and anterior column height of surgical segment. The postoperative complications were recorded. Prevertebral soft tissue edema and hydrops were assessed. The fusion rate was evaluated. The JOA improvement rate was computed at the final follow-up. Results: All the operations were successfully completed and all the patients received follow-up for (12.7±2.7) months (ranged 9-20 months). The mean operation time was (85.3±11.0) min (ranged 65-110 min). The postoperative drainage volume was (16.7±7.4) ml (ranged 5-35 ml). The JOA score and the C2-7 Cobb angle both improved at the final follow-up when compared with those before the operation (15.3±1.3 vs 12.2±2.3, 15.5°±6.1° vs 12.3°±6.0°, both P<0.001). The anterior column height of surgical segment at the final follow-up was (35.6±2.5) mm, and it was higher than that before the operation [(34.1±2.4) mm](P<0.001). No postoperative complications such as dysphagia, hoarseness, cerebrospinal fluid leakage, nerve injury, hematoma occurred. Postoperative review of cervical MRI revealed 3 cases of prevertebral soft tissue edema and hydrops without obvious symptoms. At the final follow-up, cervical spine X-ray or CT showed that all fusion segments met the criteria for osseous fusion, and the fusion rate was 100%. No complications such as neurological aggravation, internal fixation failure, fusion cage sinking, and adjacent segment degeneration was recorded at the final follow-up. At the final follow-up, the comprehensive efficacy evaluated by JOA improvement rate indicated the excellent and good rate was 90.0%(27/30): 19 cases got an excellent outcome, 8 cases got good and 3 cases got medium outcome. Conclusion: MOEA-ACDF combines the endoscopic system with ACDF technology in the treatment of CSM can achieve satisfactory clinical efficacy with high safety, and effectively restore the cervical intervertebral height and physiological curvature.

[Analysis of hemoglobin variants in Tianjin City and neighboring areas].

To determine the types and proportion of common hemoglobin variants in Tianjin and surrounding areas, to analyze the recognition ability and the effects of hemoglobin variants on experimental results in two commonly used glycated hemoglobin systems, so as to provide data support for the consistency of HbA1c detection in Tianjin City. A case-control study was used for retrospective analysis,156 specimens with abnormal electrophoretic peaks in the detection of glycated hemoglobin were collected from more than 50 000 specimens of patients in Chu Hsien-I Memorial Hospital of Tianjin Medical University between June 2020 and December 2020. Determined their hemoglobin mutation sites by DNA sequencing, and compared the values of hemoglobin variants on glycated hemoglobin detection values by high performance liquid chromatography and capillary electrophoresis. SPSS 23 was used to calculate the blood routine results of the variant specimens, and compared with the normal reference interval. The results showed that DNA sequencing identified 21 hemoglobin variants, of which 11 were α strand variants and 10 were ß strand variants. In addition, an unreported hemoglobin variant was identified, Hb Headington (HBB: c.217A>C). The HbA1c of 11 variants including Hb G-Honolulu, Hb Queens, Hb Q-Thailand, Hb J-Broussais, Hb O-Indonesia, Hb G-Coushatta, Hb G-Taipei, Hb E, Hb Headington, Hb New York and Hb D-Los Angeles were shifted by more than 7% when measured by high-performance liquid chromatography. Patients with the Hb Q-Thailand and Hb E cause reduced MCV and MCH. In conclusion, an unreported hemoglobin variant was found from Tianjin and neighboring areas. Patients with the Hb Q-Thailand and Hb E cause reduced MCV and MCH. 11 of these hemoglobin variants interfered with the detection of glycated hemoglobin using high-performance liquid chromatography, resulting in inaccurate results.

[Association study of serum LncRNA MALAT1 and SAA with type 2 diabetic kidney disease].

To investigate the correlation of serum long noncoding RNA-metastasis associated lung adenocarcinoma transcript 1(LncRNA MALAT1) and serum amyloid A(SAA) with diabetic kidney disease. Retrospective research was used, and 40 patients with type 2 diabetes and 80 patients with type 2 diabetic kidney disease patients who were treated in Tianjin Medical University Chu Hsien-I Memorial Hospital from August 2021 to February 2022 were selected, and 40 healthy subjects were selected during the same period. Reverse transcription-polymerase chain reaction(RT-PCR) was used to detect serum LncRNA MALAT1. SAA were detected with enzyme linked immunosorbent assay (ELISA). Automatic biochemistry analyzer was used to detect serum creatinine (CREA) and low-density lipoprotein cholesterol(LDL-C),automatic blood glucose analyzer to detect serum fasting plasma glucose (FPG), automatic glycated hemoglobin analyzer to detect hemoglobin A1C (HbA1c), and automatic immunoassay analyzer to detect urinary albumin to creatinine ratio(UACR). Differences between groups were compared by t test and analysis of variance. Pearson analysis was used to analyze the correlation between MALAT1, SAA and other indicators. Receiver operating characteristic curve(ROC) was used to evaluate the auxiliary diagnostic value of MALAT1 and SAA for diabetic kidney disease. The results showed that MALAT1 and SAA in the diabetic kidney disease with mass albuminuria group were higher than those in the type 2 diabetes mellitus group (q=8.57, P<0.01; q=11.09, P<0.01) and the diabetic kidney disease with microalbuminuria group (q=3.96, P<0.05; q=7.85, P<0.01). MALAT1 had a high correlation with UACR, CREA, SAA, HbA1c and FPG (r value was 0.706, 0.643, 0.578, 0.553, and 0.524, all P<0.01), and SAA had a high correlation with UACR, HbA1c and FPG (r value was 0.664, 0.617, and 0.595, all P<0.01). ROC curve analysis of the diagnostic value of LncRNA MALAT1 and protein SAA for diabetic kidney disease showed that the areas under curve (AUC) were 0.741 and 0.744, respectively. The combined diagnostic value of the two was the greatest (AUC=0.801). In summary, MALAT1 and SAA were elevated in the serum of patients with type 2 diabetes. Their concentrations in the serum of group with diabetic kidney disease were higher than that in the type 2 diabetes group, and the serum concentrations of MALAT1 and SAA in group with mass albuminuria are higher than the group with microalbuminuria. MALAT1 and SAA were both closely related to UACR and HbA1c, and there is a correlation between them. Both of them may have ancillary diagnostic value for diabetic kidney disease.

[Genetics Analysis of patients with Dravet syndrome due to mosaicism variation of paternal SCN1A gene].

Genetic analysis was performed on a family of fraternal twins affected with Dravet syndrome by genetic tests whose parents were normal. To further analyze the cause of the disease, the fraternal twins were subjected to whole exome sequencing (WES), and the family was verified by Sanger sequencing, with the father semen and peripheral blood DNA were further analysed by target sequencing. The WES test identified a heterozygous c.5348C>T (p.Ala1783Val) variant of the SCN1A gene in the fraternal twins, which was predicted to be pathogenic and was detected in the father peripheral blood and semen, but not in the mother. So the mosaicism mutation of paternal SCN1A gene might be the genetic cause of Dravet syndrome in offspring.

[Deleted in lymphocytic leukemia 1 promoted proliferation and apoptosis of nephroblastoma cells through regulating miR-513a-5p and RANBP2 pathway].

Objective: To study the regulatory effects and mechanisms of deleted in lymphocytic leukemia 1 (DLEU1), microRNA-513a-5p (miR-513a-5p), and RAN binding protein 2 (RANBP2) in nephroblastoma. Methods: The GHINK-1 cells were transfected with pcDNA (pcDNA group), pcDNA-DLEU1 (pcDNA-DLEU1 group), miR-NC (miR-NC group), miR-513a-5p mimics (miR-513a-5p group), pcDNA-RANBP2 (pcDNA-RANBP2 group), pcDNA-DLEU1 and miR-NC (pcDNA-DLEU1+ miR-NC group), pcDNA-DLEU1 and miR-513a-5p mimics (pcDNA-DLEU1+ miR-513a-5p group), miR-513a-5p mimics and pcDNA (miR-513a-5p+ pcDNA group), miR-513a-5p mimics and pcDNA-RANBP2 (miR-513a-5p + pcDNA-RANBP2 group). Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to detect the expressions of DLEU1, miR-513a-5p, RANBP2 in nephroblastoma tissues, normal adjacent tissues, normal kidney cell HK2, and hemangioblastoma cell GHINK-1. Western blot was used to detect the expressions of proliferating cell nuclear antigen (PCNA), B cell lymphoma/leukemia-2 (Bcl-2) and Bcl-2 related X (Bax). Cell counting kit 8 (CCK-8) was used to detect the cell survival rate. Flow cytometry was used to detect the apoptosis rate. Dual luciferase report test was used to detect the luciferase activity of cells. Results: The expression levels of DLEU1, miR-513a-5p and RANBP2 in adjacent tissues were 1.02±0.08, 1.01±0.06, 1.00±0.05, respectively, significantly lower than 5.16±0.24, 0.23±0.02, 1.67±0.09 in nephroblasts tumor tissues (P<0.05). Their expression levels in HK2 cells were 1.00±0.06, 1.00±0.08, 1.02±0.09, respectively, significantly lower than 3.15±0.21, 0.18±0.01, 1.54±0.10 in GHINK-1 cells (P<0.05). Overexpression of DLEU1 significantly reduced the apoptosis rate (7.35±0.41 vs 12.35±1.12, P<0.05). Overexpression of RANBP2 significantly reduced the apoptosis rate (8.89±0.48 vs 12.64±1.12, P<0.05). Compared with the miR-NC group (1.01±0.06, 0.99±0.06), the luciferase activity of DLEU1-WT (0.43±0.04) and RANBP2-WT (0.61±0.07) in miR-513a-5p group were significantly reduced (P<0.05). Compared with anti-miR-NC group (0.99±0.07, 0.98±0.05), the luciferase activity of DLEU1-WT (1.34±0.11) and RANBP2-WT (1.39 ±0.13) in anti-miR-513a-5p group was significantly increased (P<0.05). Simultaneous overexpression of pcDNA-DLEU1 and miR-513a-5p in GHINK-1 cells significantly reduced the apoptosis rate (11.34±1.03 vs 8.51±0.69, P<0.05). Simultaneous overexpression of miR-513a-5p and RANBP2 in GHINK-1 cells significantly reduced the apoptosis rate (9.96±0.72 vs 15.94±1.00, P<0.05). Conclusions: The long-chain non-coding RNA (lncRNA) DLEU1 can promote the proliferation and inhibit the apoptosis of nephroblastoma cells. The mechanism is related to the targeted regulation of miR-513a-5p and RANBP2 function, which will provide theoretical support for the nephroblastoma treatment.

[Monitoring metrics for short-term exposure to ambient ozone and pulmonary function and airway inflammation in healthy young adults].

OBJECTIVE: To assess the associations of different monitoring metrics for short-term exposure to ambient ozone (O3) with pulmonary function and airway inflammation in healthy young adults. METHODS: A total of 97 healthy young college students were recruited and followed in a panel study conducted from December 2017 to June 2018. Each participant underwent 3 follow-up visits, and lung function and fractional exhaled nitric oxide (FeNO) were measured at each visit. Ambient air pollutant concentrations were obtained from the environment monitoring station of Beijing closest to the participant residences, and meteorological data were collected from China Meteorological Data Service Center. Linear mixed-effect models were applied to assess the associations between different monitoring metrics for ambient O3 short-term exposure with pulmonary function or airway inflammation in the healthy young adults. RESULTS: During the study period, the P50 (P25, P75) values for ambient O3 concentration expressed as daily 1-hour maximum (O3-1 h max), daily maximum 8-hour average (O3-8 h max) and 24-hour average (O3-24 h avg) were 102.5 (76.8, 163.0) µg/m3, 91.1 (68.3, 154.3) µg/m3 and 61.6 (36.9, 81.7) µg/m3, respectively. The different monitoring metrics for short-term exposure to ambient O3 were significantly associated with reduced forced expiratory volume in the first second (FEV1) and increased FeNO. An interquartile range (IQR) increase in 6-d moving average of O3-1 h max (IQR=71.5 µg/m3) was associated with a 6.2% (95%CI: －11.8%, －0.5%) decrease in FEV1 and a 63.3% (95%CI: 13.8%, 134.3%) increase in FeNO. An IQR increase in 7-d moving average of O3-8 h max (IQR=62.0 µg/m3) was associated with a 6.2% (95%CI: －11.6%, －0.7%) decrease in FEV1and a 75.5% (95%CI: 19.3%, 158.0%) increase in FeNO. An IQR increase in 5-d moving average of O3-24 h avg (IQR=32.9 µg/m3) was associated with a 3.7% (95%CI: －7.1%, －0.2%) decrease in FEV1and a 25.3% (95%CI: 3.6%, 51.6%) increase in FeNO. There was no significant association between the three monitoring metrics for O3 exposure and peak expiratory flow (PEF). CONCLUSION: Short-term exposure to ambient O3 was associated with decreased lung function and increased airway inflammation among the healthy young adults, and daily 1-hour maximum was more sensitively to the respiratory effects of O3.

microRNA-935 is reduced in non-small cell lung cancer tissue, is linked to poor outcome, and acts on signal transduction mediator E2F7 and the AKT pathway.

BACKGROUND: A potential role for microRNA-935 (miR-935) has been identified in several cancers but not in non-small cell lung cancer (NSCLC). We hypothesised changes in miR-935 in NSCLC, and proposed mechanisms that may further explain its role in carcinogenesis. METHODS: NSCLC tissue and nearby normal tissue was obtained from 101 patients and was probed by qRT-PCR for miR-935 expression. The role of miR-935 and a potential target (signal transduction factor E2F7) was determined in cell lines by a dual luciferase assay. The function of miR-935 was investigated through metabolic activity (MTT) and transwell migration assays. Western blot and immunocytochemical assays examined protein expression level. Growth of miR-935 transfected or untransfected cells was measured via xenograft tumour formation. RESULTS: miR-935 was reduced in cancer tissue and was related to lymph node metastases, tumour node metastasis status and poor prognosis (all p < 0.02). In vitro, miR-935 suppressed cell proliferation, migration and invasion in NSCLC cells through targeting E2F7. Furthermore, E2F7 was upregulated in NSCLC tissue associated with poor prognosis (p = 0.0203) of NSCLC patients. miR-935 suppressed the epithelial-mesenchymal transition and AKT pathways in NSCLC and inhibited the tumour growth in vivo. CONCLUSION: Altered miR-935 in lung cancer biopsy tissue may be a diagnostic tool and could direct treatment. Involvement in carcinogenesis is implied by its suppression of the development of NSCLC via targeting E2F7 and inhibiting AKT pathway.

Liraglutide ameliorates palmitate-induced insulin resistance through inhibiting the IRS-1 serine phosphorylation in mouse skeletal muscle cells.

OBJECTIVE: A reduction in insulin-stimulated glucose uptake in skeletal muscles is a characteristic of insulin resistance and type 2 diabetes mellitus (T2DM). The glucagon-like peptide (GLP)-1 agonist liraglutide can reduce blood glucose levels in individuals with T2DM. However, its effect on insulin-induced glucose metabolism in the skeletal muscle of insulin resistance is unknown. We investigated the effects and action mechanisms of liraglutide on insulin resistance (IR) in the skeletal muscle cells treatment with palmitic acid (PA). METHODS: The cell-surface GLUT4myc levels were determined by an antibody-coupled colorimetric assay. The phosphorylation levels of Akt, PI3K(p85α), AS160, IRS1, IKK, and JNK were determined by western blotting. The quantifications of mRNA levels of TNFα, IL-1ß, and IL-6 were determined by real-time PCR. Analysis of variance was used for data analysis. RESULTS: PA elevated not only phosphorylation of JNK, IRS1 serines, and IKKα/ß, but also the expression of IL-6, TNFα and IL-1ß in C2C12-GLUT4myc cells. PA can reduce phosphorylation of IRS1 tyrosine. These effects of PA were reversed by liraglutide. In addition, liraglutide can reverse PA-decreased insulin-stimulated cell-surface GLUT4 levels, Akt, PI3K(p85α), and AS160 phosphorylation. CONCLUSIONS: Liraglutide can enhance insulin-induced GLUT4 translocation by inhibiting IRS1 serine phosphorylation in PA-treated muscle cells.

[Effects of equol on colon cancer cell proliferation].

OBJECTIVE: To investigate the effect of equol on the proliferation of colom cancer cells and to explore the mechanisms. METHODS: Colon cancer cells (DLD1,HCT15,COLO205,LOVO,SW480) were incubated, the cell proliferation was identified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay. Reverse transcription PCR and Western blot were used to measure the mRNA and the protein expression of estrogen receptor and nuclear factor (erythroid-derived 2)-like 2 (Nrf2)in the colon cancer cells, respectively. Moreover, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay was used to investigate the effect of estrogen receptor(ER) inhibitor,ERα agonist, and estrogen receptor ERßagonist on the cell proliferation. RESULTS: ERα was faintly expressed in the DLD-1 and HCT-15 cells. However, ERß expression in DLD1, HCT15, COLO205, LOVO, and SW480 colon cancer cells. Different concentrations of equol (0, 0.5, 1, 5, 10 µmol/L) significantly inhibited the growth of HCT-15 cell with the expression of ERα and ERß.More-over, different concentrations of equol (0, 0.5, 1, 5, 10 µmol/L) significantly inhibited the growth of LOVO, and SW480 cells with the ERß expression in a dose-dependent manner as demonstrated with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay. mRNA expressions of ERα and ERß in HCT-15 were stimulated significantly. Western blotting proved that the protein expressions of ERα and ERß increased with the increasing of equol dose. Moreover we found significant difference of Nrf2 protein expression in HCT-15 cell stimulated by different concentrationss of equol. After the similation of estrogen receptor inhibitor, ERα agonist, or ERß agonist, we found that only dif-ferent concentrations of ERß agonist(0, 1, 10, 100, 1 000, 10 000 nmol/L) significantly inhibited the growth of HCT-15, LOVO, and SW480 in adose-dependent manner. Estrogen receptor inhibitor and ERα agonistdid not present significant effect on the cell proliferation of HCT-15, LOVO, and SW480. CONCLUSION: Equol inhibited the colon cancer cell proliferation by its estrogenic activities and antioxidant activities.

[Research advances in the role of mesenchymal stem cells and their exosomes in treatment of liver diseases].

At present, liver transplantation remains the most effective treatment for acute liver failure and advanced cirrhosis, but its use and promotion are limited by insufficient organ donors, financial consideration, and requirements for lifelong immunosuppression. In recent years, stem cell transplantation has been recommended as an effective substitutive therapy for liver disease. Mesenchymal stem cells, also known as pluripotent interstitial stromal cells, are self-renewing cells that can be found in almost all postnatal organs and tissues, including the liver. Their potential to differentiate into hepatocytes and immunomodulatory properties provide new insights into the use of mesenchymal stem cells in the treatment of acute and chronic liver diseases. This article reviews the characteristics of mesenchymal stem cells, their mechanisms in the treatment of acute and chronic liver diseases, and related risks.

Predictive value of CK20 in evaluating the efficacy of treatment and prognosis after surgery for colorectal cancer.

Herein, correlations between expression levels of CK20 and efficacy of treatment and postoperative prognosis of colorectal cancer were evaluated to elucidate the clinical value of CK20. Postoperative follow-up was performed on 62 patients who underwent surgery for colorectal cancer between January 2010 and December 2010. Samples of tumor tissues and intraperitoneal drainage fluids were collected. Blood samples were obtained during the 2-year follow-up period. The expression of CK20 in surgical specimen, intraperitoneal drainage fluids, and postoperative serum samples was quantified by enzyme-linked immunosorbent assay, RT-PCR, and western blotting. Correlation between the levels of CK20 and postoperative outcomes was investigated by Spearman correlation analysis. In both tumor specimens and intraperitoneal drainage fluids, CK20 levels were lower in patients with earlier cancer stages than in those at later stages. During postoperative follow-up, serum negative CK20 patients had significantly higher 3-year survival rates than serum positive CK20 patients. All differences were statistically significant (P < 0.05). CK20 levels can provide clinically valuable information on the postoperative prognosis of patients with colorectal cancer.

Isolation and characterization of novel microsatellite markers from the sika deer (Cervus nippon) genome.

Microsatellite markers are widely and evenly distributed, and are highly polymorphic. Rapid and convenient detection through automated analysis means that microsatellite markers are widely used in the construction of plant and animal genetic maps, in quantitative trait loci localization, marker-assisted selection, identification of genetic relationships, and genetic diversity and phylogenetic tree construction. However, few microsatellite markers remain to be isolated. We used streptavidin magnetic beads to affinity-capture and construct a (CA)n microsatellite DNA-enriched library from sika deer. We selected sequences containing more than six repeats to design primers. Clear bands were selected, which were amplified using non-specific primers following PCR amplification to screen polymorphisms in a group of 65 unrelated sika deer. The positive clone rate reached 82.9% by constructing the enriched library, and we then selected positive clones for sequencing. There were 395 sequences with CA repeats, and the CA repeat number was 4-105. We selected sequences containing more than six repeats to design primers, of which 297 pairs were designed. We next selected clear bands and used non-specific primers to amplify following PCR amplification. In total, 245 pairs of primers were screened. We then selected 50 pairs of primers to randomly screen for polymorphisms. We detected 47 polymorphic and 3 monomorphic loci in 65 unrelated sika deer. These newly isolated and characterized microsatellite loci can be used to construct genetic maps and for lineage testing in deer. In addition, they can be used for comparative genomics between Cervidae species.

Effects of rare earth element lanthanum on rumen methane and volatile fatty acid production and microbial flora in vitro.

The objectives of the trial were to study the effects of rare earth element (REE) lanthanum (La) on the in vitro rumen methane (CH4 ) and volatile fatty acid (VFA) production and the microbial flora of feeds. Four feed mixtures with different levels of neutral detergent fibre (NDF), that is 20.0% (I), 31.0% (II), 41.9% (III) and 52.7% (IV), were formulated as substrates. Five levels of LaCl3 , that is 0, 0.4, 0.6, 0.8 and 1.0 mmol/kg dry matter (DM), were added to the feed mixtures, respectively, as experimental treatments in a two-factor 5 × 4 randomized design. The in vitro incubation lasted for 24 h. The results showed that supplementing LaCl3 increased the total gas (p < 0.001) production and tended to increase the total VFA production (p = 0.072) and decreased the CH4 production (p = 0.001) and the ratios of acetate/propionate (p = 0.019) and CH4 /total VFA (p < 0.001). Interactions between LaCl3 and NDF were significant in total gas production (p = 0.030) and tended to be significant in CH4 production (p = 0.071). Supplementing LaCl3 at the level of 0.8 mmol/g DM decreased the relative abundance of methanogens and protozoa in the total bacterial 16S rDNA analysed using the real-time PCR (p < 0.0001), increased F. succinogenes (p = 0.0003) and decreased R. flavefaciens (p < 0.0001) whereas did not affect R. albus and anaerobic fungi (p > 0.05). It was concluded that LaCl3 decreased the CH4 production without negatively affecting feed digestion through manipulating rumen microbial flora when feed mixtures with different levels of NDF were used as substrates.

Virtual classroom helps medical education for both Chinese and foreign students.

AIMS: The rapid development of computer and internet technology has a strong influence over one's quality of education within different fields of study. To determine the potential benefits of introducing internet into medical school classes, a pilot study was conducted in three different Chinese medical schools. MATERIALS AND METHODS: Seven hundred and eight medical school undergraduates, 385 dental school students and 366 foreign students were randomly recruited to complete a self-administered questionnaire. The contents included personal information, current usage of computer and internet, and attitudes towards the computerised teaching methods. Two forum groups were created using instant message software and were randomly assigned to two classes, allowing students to freely ask or discuss questions with the help of their teachers in these two virtual classrooms. RESULTS: All 1539 questionnaires were accepted and analysed. Although there were some differences between Chinese and foreign undergraduates, both group of students were highly proficient in internet usage and navigation. Overwhelmingly, 88.37% of the students owned a computer and frequently logged onto the internet. Most of them believed that the internet is a helpful adjunct to their studies and held positive attitudes towards computerised teaching. Compared to the classes that were not assigned internet forums, the two experimental classes performed significantly better on the examination. CONCLUSIONS: Our results suggest that computerised teaching methods have significant potential to assist in learning for both Chinese and foreign medical undergraduates.

Ca²âº signals promote GLUT4 exocytosis and reduce its endocytosis in muscle cells.

Elevating cytosolic Ca(2+) stimulates glucose uptake in skeletal muscle, but how Ca(2+) affects intracellular traffic of GLUT4 is unknown. In tissue, changes in Ca(2+) leading to contraction preclude analysis of the impact of individual, Ca(2+)-derived signals. In L6 muscle cells stably expressing GLUT4myc, the Ca(2+) ionophore ionomycin raised cytosolic Ca(2+) and caused a gain in cell surface GLUT4myc. Extra- and intracellular Ca(2+) chelators (EGTA, BAPTA-AM) reversed this response. Ionomycin activated calcium calmodulin kinase II (CaMKII), AMPK, and PKCs, but not Akt. Silencing CaMKIIδ or AMPKα1/α2 partly reduced the ionomycin-induced gain in surface GLUT4myc, as did peptidic or small molecule inhibitors of CaMKII (CN21) and AMPK (Compound C). Compared with the conventional isoenzyme PKC inhibitor Gö6976, the conventional plus novel PKC inhibitor Gö6983 lowered the ionomycin-induced gain in cell surface GLUT4myc. Ionomycin stimulated GLUT4myc exocytosis and inhibited its endocytosis in live cells. siRNA-mediated knockdown of CaMKIIδ or AMPKα1/α2 partly reversed ionomycin-induced GLUT4myc exocytosis but did not prevent its reduced endocytosis. Compared with Gö6976, Gö6983 markedly reversed the slowing of GLUT4myc endocytosis triggered by ionomycin. In summary, rapid Ca(2+) influx into muscle cells accelerates GLUT4myc exocytosis while slowing GLUT4myc endocytosis. CaMKIIδ and AMPK stimulate GLUT4myc exocytosis, whereas novel PKCs reduce endocytosis. These results identify how Ca(2+)-activated signals selectively regulate GLUT4 exocytosis and endocytosis in muscle cells.

[Peripheral blood cell count composite score as a prognostic factor in patients with colorectal cancer].

Objective: To develop a prognostic prediction model for patients with colorectal cancer based on a peripheral blood cell composite score (PBCS) system. Methods: This retrospective observational study included patients who had primary colorectal cancer without distant metastasis, who did not undergo radiotherapy or chemotherapy before surgery, who did not receive leukocyte or platelet-raising therapy within 1 month before surgery, and whose postoperative pathology confirmed colorectal adenocarcinoma with complete tumor resection. Patients with severe anemia, infection, or hematologic diseases before surgery, as well as those with severe heart, lung, or other important organ diseases or concurrent malignant tumors, were excluded. In total, 1021 patients with colorectal cancer who underwent surgical treatment in the Department of Gastrointestinal Surgery of the Fourth Hospital of Hebei Medical University from April 2018 to April 2020 were retrospectively included as the training set (766 patients) and the internal validation set (255 patients). Additionally, using the same criteria, 215 patients with colorectal cancer who underwent surgical treatment in another treatment group from March 2015 to December 2020 were selected as the external validation set. The "surv_cutpoint" function in R software was used to analyze the optimal cut-off values of neutrophils, lymphocytes, and platelets, and a PBCS system was established based on the optimal cut-off values. The scoring rules of the PBCS system were as follows: Neutrophils and platelets below the optimal cut-off value = 1 point, otherwise 0 points; Lymphocytes above the optimal cut-off value = 1 point, otherwise 0 points. The scores of the three cell types were added together to obtain the PBCS. Univariate and multivariate Cox regression analyses were performed to explore the correlation between patients' clinicopathological features and prognosis, and a nomogram was constructed based on the Cox regression analysis to predict patients' prognosis. The accuracy of the nomogram prediction model was validated using the C-index, calibration curve, and decision curve analysis. Results: The optimal cut-off values for neutrophils, lymphocytes, and platelets were 4.40×109/L, 1.41×109/L, and 355×109/L, respectively. The patients were divided into high and low groups according to the optimal cut-off values of these cells. Survival curve analysis showed that a high lymphocyte count (training set: P=0.042, internal validation: P=0.010, external validation: P=0.029), low neutrophil count (training set: P=0.035, internal validation: P=0.001, external validation: P=0.024), and low platelet count (training set: P=0.041, internal validation: P=0.030, external validation: P=0.024) were associated with prolonged overall survival (OS), with statistically significant differences in all cases. Survival analysis of different PBCS groups showed that patients with a high PBCS had longer OS than those with a low PBCS (P<0.05). Univariate and multivariate Cox regression analysis results showed that aspirin use history, vascular thrombus, neural invasion, CA19-9, N stage, operation time, M stage, and PBCS were independent factors affecting OS (all P<0.05). The PBCS was also an independent factor affecting disease-specific survival (P<0.05), but not progression-free survival (P>0.05). The above independent risk or protective factors were included in R software to construct a nomogram for predicting OS. The C-index (0.873), calibration curve, and decision curve analysis (threshold probability: 0.0%-75.2%) all indicated that the nomogram prediction model had good predictive performance for OS. Conclusion: This study demonstrates that the PBCS constructed based on preoperative peripheral blood levels of neutrophils, lymphocytes, and platelets is an independent factor associated with the prognosis of patients with colorectal cancer. The nomogram model constructed based on this score system exhibits good predictive efficacy for the prognosis of these patients.

The diagnostic value of serum procalcitonin, IL-10 and C-reactive protein in community acquired pneumonia and tuberculosis.

AIM: To explore the diagnostic value of serum procalcitonin (PCT), interleukin-10 (IL-10), C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) in community acquired pneumonia (CAP) and tuberculosis (PTB). PATIENTS AND METHODS: 113 CAP cases patients and 78 PTB cases were enrolled from May 2011 to March 2012. Routine blood test, serum PCT, CRP, IL-10 and ESR of patients within 24 hours were analyzed retrospectively. RESULTS: The serum concentrations of PCT, IL-10, CRP and ESR in CAP patients with CAP were 0.35±0.017 mg/mL, 0.095±0.004 mg/L, 59.80±5.12 mg/L and 35.00±4.81 mm/1h, respectively, significantly higher than patients with PTB (p < 0.01); According to the result of ROC curve analysis in CAP and PTB, the PTC area under ROC curve is 0.715 (95% CI 0.647-0.782), the sensitivity and specific degree of serum PTC were significant better than CRP and IL10 (p < 0.05). In tuberculosis sputum culture, the serum concentrations of IL-10 and ESR in TB positive group were 0.045±0.013 mg/L and 62.50±8.69 mm/1h, significantly higher than that of TB negative group (p < 0.05); whereas, the concentrations of serum PCT and CRP in TB positive and negative groups had no significant difference (p > 0.05). CONCLUSIONS: The levels of serum PCT, IL-10, CRP and ESR in CAP patients are higher than that in PTB patients. Therefore, the serum PCT, IL10, CRP and ESR level is benefit to distinguish between CAP and PTB. This could provide a comprehensible evidence for both diagnosis and prognosis.