Construction of TRAF6 ubiquitin site 331 mutant colorectal cancer cell stable line and its effect on biological behavior of colorectal cancer cells / 中华肿瘤杂志

Objective: To investigate the effect of ubiquitin mutation at position 331 of tumor necrosis factor receptor related factor 6 (TRAF6) on the biological characteristics of colorectal cancer cells and its mechanism. Methods: lentivirus wild type (pCDH-3×FLAG-TRAF6) and mutation (pCDH-3×FLAG-TRAF6-331mut) of TRAF6 gene expression plasmid with green fluorescent protein tag were used to infect colorectal cancer cells SW480 and HCT116, respectively. The infection was observed by fluorescence microscope, and the expressions of TRAF6 and TRAF6-331mut in cells was detected by western blot. Cell counting kit-8 (CCK-8) and plate cloning test were used to detect the proliferation ability of colorectal cancer cells in TRAF6 group and TRAF6-331mut group, cell scratch test to detect cell migration, Transwell chamber test to detect cell migration and invasion, immunoprecipitation to detect the ubiquitination of TRAF6 and TRAF6-331mut with ubiquitinof lysine binding sites K48 and K63. Western blot was used to detect the effects of TRAF6 and TRAF6-331mut over expression on the nuclear factor kappa-B (NF-κB) and mitogen activated protein kinase mitogen-activated protein kinase (MAPK)/activating protein-1(AP-1) signal pathway. Results: The successful infection of colorectal cancer cells was observed under fluorescence microscope. Western blot detection showed that TRAF6 and TRAF6-331mut were successfully expressed in colorectal cancer cells. The results of CCK-8 assay showed that on the fourth day, the absorbance values of HCT116 and SW480 cells in TRAF6-331mut group were 1.89±0.39 and 1.88±0.24 respectively, which were lower than those in TRAF6 group (2.09±0.12 and 2.17±0.45, P=0.036 and P=0.011, respectively). The results of plate colony formation assay showed that the number of clones of HCT116 and SW480 cells in TRAF6-331mut group was 120±14 and 85±14 respectively, which was lower than those in TRAF6 group (190±21 and 125±13, P=0.001 and P=0.002, respectively). The results of cell scratch test showed that after 48 hours, the percentage of wound healing distance of HCT116 and SW480 cells in TRAF6-331mut group was (31±12)% and (33±14)%, respectively, which was lower than those in TRAF6 group [(43±13)% and (43±7)%, P=0.005 and 0.009, respectively]. The results of Transwell migration assay showed that the migration numbers of HCT116 and SW480 cells in TRAF6-331mut group were significantly lower than those in TRAF6 group (P<0.001 and P<0.002, respectively). The results of Transwell invasion assay showed that the number of membrane penetration of HCT116 and SW480 cells in TRAF6-331mut group was significantly lower than those in TRAF6 group (P=0.008 and P=0.009, respectively). The results of immunoprecipitation detection showed that the ubiquitin protein of K48 chain pulled by TRAF6-331mut was lower than that of wild type TRAF6 in 293T cells co-transfected with K48 (0.57±0.19), and the ubiquitin protein of K63 chain pulled down by TRAF6-331mut in 293T cells co-transfected with K63 was lower than that of wild type TRAF6 (0.89±0.08, P<0.001). Western blot assay showed that the protein expression levels of NF-κB, p-NF-κB and p-AP-1 in TRAF6-331mut-HCT116 cells were 0.63±0.08, 0.42±0.08 and 0.60±0.07 respectively, which were lower than those in TRAF6-HCT116 cells (P=0.002, P<0.001 and P<0.001, respectively). The expression level of AP-1 protein in TRAF6-HCT116 cells was 0.89±0.06, compared with that in TRAF6-HCT116 cells. The difference was not statistically significant (P>0.05). The protein expression levels of NF-κB, p-NF-κB and p-AP-1 in TRAF6-331mut-SW480 cells were 0.50±0.06, 0.51±0.04, 0.48±0.02, respectively, which were lower than those in TRAF6-SW480 cells (all P<0.001). There was no significant difference in AP-1 protein expression between TRAF6-331mut-SW480 cells and TRAF6-SW480 cells. Conclusion: The ubiquitin site mutation of TRAF6 gene at 331 may prevent the binding of TRAF6 and ubiquitin lysine sites K48 and K63, and then affect the expressions of proteins related to downstream NF-κB and MAPK/AP-1 signal pathways, and inhibit the proliferation, migration and invasion of colorectal cancer cells.

Epithelial Cell Adhesion Molecules (EpCAMs) Promote Drug Resistance in Breast Cancer Cells by Enhancing Tight Junctions / 中国生物化学与分子生物学报

Breast cancer is a malignant tumor with high mortality, and multidrug resistance (MDR) mediated by ABCG2 (ATP-Binding cassette G2) is an important cause of chemotherapy failure. It is an urgent problem to explore the mechanism of ABCG2-mediated drug resistance and its key molecules. Epithelial cell adhesion molecule (EpCAM) is involved in multiple tumor drug resistance and is closely related to breast cancer MDR. However, its role in ABCG2-mediated breast cancer drug resistance has not been clarified. The purpose of this study was to explore the regulation of EpCAM on ABCG2-mediated MDR in breast cancer cells and its mechanism. CCK8 cytotoxicity assays confirmed that the drug resistance of MCF-7/MX cell line to mitoxantrone (MX) was significantly increased compared with MCF-7 drug-sensitive strain of human breast cancer. Western blotting results showed that ABCG2 was highly expressed and EpCAM was up-regulated in MCF-7/MX cells compared with MCF-7. SiRNA knockdown of EpCAM in MCF-7/MX cells down-regulated ABCG2 expression and restored sensitivity to MX. Cell morphology was observed under an inverted microscope, and it was found that knocking down EpCAM reduced cell-cell connections between MCF-7/MX cells. The co-localization of EpCAM and claudin 1 in MCF-7/MX cells was observed by immunofluorescence. Furthermore, Western blotting results showed that EpCAM knockdown reduced claudin 1 expression in MCF-7/MX cells. In conclusion, EpCAM may promote ABCG2-mediated mMDR in breast cancers by enhancing intercellular tight junctions through interaction with claudin 1.

Cloning and functional analysis of the phenylalaninammo-nialyase gene from Rhododendron fortunei / 生物工程学报

Phenylalaninammo-nialyase (PAL) is a key enzyme in the synthesis of methyl benzoate - a plant aroma compound. In order to understand the function of this enzyme in the formation of fragrance in the scented Rhododendron species-Rhododendron fortunei, we cloned a gene encoding this enzyme and subsequently examined the gene expression patterns and the profile of enzyme activity during development in various tissues. The full length of RhPAL gene was cloned by reverse transcription-PCR (RT-PCR) and rapid amplification of cDNA ends (RACE) techniques. The expression levels of RhPAL gene were measured by real-time quantitative reverse transcription PCR (qRT-PCR) and the amount of phenylalanine and cinnamic acid were assayed with LC-MS. The results showed that the ORF sequence of RhPAL gene amplified from the cDNA templates of flower buds had 2 145 bp, encoding 715 amino acids, and shared 90% homology to the PAL amino acid sequences from other species. qRT-PCR analysis showed that the expression of RhPAL in petals during flowering kept in rising even until the flowers wilted. The expression of RhPAL in pistil was much higher than that in stamen, while the expression in the younger leaves was higher than in old leaves. However, the expression level was relatively lower in petal and stamen compared to that in leaves. We also measured the PAL activity by Enzyme-linked immuno sorbent assay in the petals of flowers at different flowering stages. The results showed that PAL activity reached the highest at the bud stage and then decreased gradually to the lowest when the flowers wilted, which followed a similar trend in the emission of the flower fragrance. The phenylalanine and cinnamic acid contents measured by LC-MS were highly correlated to the expression level of RhPAL in various tissues and at different flowering stages, implying that RhPAL plays an important role in the formation of the flower fragrance. This work may facilitate the breeding and improvement of new fragrant Rhododendron cultivars.

Renin-angiotensin system inhibitor is associated with the reduced risk of all-cause mortality in COVID-19 among patients with/without hypertension / 医学前沿

Consecutively hospitalized patients with confirmed coronavirus disease 2019 (COVID-19) in Wuhan, China were retrospectively enrolled from January 2020 to March 2020 to investigate the association between the use of renin-angiotensin system inhibitor (RAS-I) and the outcome of this disease. Associations between the use of RAS-I (angiotensin-converting enzyme inhibitor (ACEI) or angiotensin receptor blocker (ARB)), ACEI, and ARB and in-hospital mortality were analyzed using multivariate Cox proportional hazards regression models in overall and subgroup of hypertension status. A total of 2771 patients with COVID-19 were included, with moderate and severe cases accounting for 45.0% and 36.5%, respectively. A total of 195 (7.0%) patients died. RAS-I (hazard ratio (HR)= 0.499, 95% confidence interval (CI) 0.325-0.767) and ARB (HR = 0.410, 95% CI 0.240-0.700) use was associated with a reduced risk of all-cause mortality among patients with COVID-19. For patients with hypertension, RAS-I and ARB applications were also associated with a reduced risk of mortality with HR of 0.352 (95% CI 0.162-0.764) and 0.279 (95% CI 0.115-0.677), respectively. RAS-I exhibited protective effects on the survival outcome of COVID-19. ARB use was associated with a reduced risk of all-cause mortality among patients with COVID-19.

Screening of Differential Genes for Endometriosis and Expression Validation of the Potential Target GAS6 / 中国生物化学与分子生物学报

Growth arrest specific protein 6 (GAS6) plays an important role in the occurrence and development of tumors, and its signal transduction is involved in cell proliferation, adhesion and migration, but its related functions and molecular mechanisms in endometriosis (EMs) are still unclear. In this study, we searched and downloaded the transcriptome datasets of EMs from GEO database and performed GEO online analysis, and then screened out the differentially expressed genes and performed cluster analysis based on GO and KEGG pathway. The mRNA levels of the differentially expressed genes shared by more than three datasets were verified by qRT-PCR in the endometrium of ten women with no endometriosis and no clear disease and the ectopic endometrium of 11 patients with ovarian chocolate cysts. Immunohistochemistry and qRT-PCR were used to verify the expression of GAS6 and epithelial mesenchymal transition (EMT) marker genes, and immunofluorescence was used to co-label GAS6 and E-cadherin in endometriosis clinical samples. In this study, a total of 47 differentially expressed genes were screened out of the four transcriptome datasets, which were mainly enriched in processes such as cell migration and related signal pathways such as MAPK, PI3K-AKT, and tight junction. The mRNA levels of the nine differentially expressed genes shared by more than three datasets in endometriosis patients were consistent with the results of bioinformatics analysis. GAS6 expression levels in ectopic endometrium of EMs patients are higher than the control group (P < 0. 05), and EMs patients have the characteristics of EMT in the ectopic endometrial tissue, that is, the expression of E-cadherin is down-regulated (P < 0. 05) and the expression of vimentin is up-regulated (P < 0. 01). The expression of E-cadherin in the ectopic endometrial glandular epithelial cells of EMs patients is low while the expression of GAS6 is up-regulated, suggesting that GAS6 may mediate the EMT process in endometriosis. In conclusion, this study reveals that GAS6 is highly expressed in endometriosis patients and may mediate the EMT process to participate in the occurrence and development of endometriosis, providing a potential target for clinical treatment of endometriosis.

Epithelial cell adhesion molecule promotes metastasis and multidrug resistance of breast cancer through induction of epithelial-mesenchymal transition / 中国药理学通报

Aim: To investigate the effect of epithelial cell adhesion molecule (EpCAM) on metastasis and multidrug resistance of breast cancer and its mechanism. Methods Immunohistochemical staining was employed to detect the expression of EpCAM in adjacent non-tumor tissues (ANTTs) and breast cancer tissues. siRNA was applied to knock down EpCAM expression in MDA-MB-231 cells. Transwell assay was used to detect the invasion and migration ability of breast cancer cells. Western blot analysis was performed to determine the protein expression of EpCAM, epithelial mesenchymal transition (EMT) markers E-cadherin, N-cadherin and vimentin, breast cancer resistance protein (BCRP), and β-catenin. Results EpCAM immunoreactivity was consistently stronger in primary breast cancer tissues and even higher in metastatic lesions than that in ANTTs. The expression of EpCAM was significantly upregulated in triple negative breast cancer MDA-MB-231 cells. EpCAM knockdown using siRNA decreased the invasion and migration ability and BCRP expression, and partially reversed the EMT phenotypes of MDA-MB-231 cells, β-catenin expression was upregulated in MDA-MB-231 cells. ICG-001, a specific Wnt/β-catenin pathway inhibitor, downregulated the expression levels of EpCAM, N-cadherin, and vimentin in MDA-MB-231 cells. Conclusions EpCAM could promote metastasis and drug resistance of breast cancer through the induction of EMT, which is related to the Wnt/β-catenin signaling pathway.

Effects of ultrafine particulates on cardiac function in rat isolated heart / 北京大学学报(医学版)

OBJECTIVE@#To evaluate whether ultrafine particulates (UFPs) have direct deleterious effects on cardiac function through activating MAPK signaling.@*METHODS@#Langendorff-perfused Sprague-Dawley rat hearts were randomly divided into 2 groups (n=10/each group). In control group, the rat hearts were perfused with Tyrode's buffer for 40 min; in UFPs-treated group, the hearts were perfused with UFPs at a concentration of 12.5 mg/L. Cardiac function was determined by measuring left ventricular developed pressure (LVDP), left ventricular peak rate of contraction and relaxation (±dp/dtmax) and coronary flow (CF). The levels of malondialdehyde (MDA), superoxide dismutase (SOD), total anti-oxidant capacity (TAOC) were detected in order to evaluate cardiac oxidative stress via the thiobarbituric acid assay, water soluble tetrazolium salt assay and colorimetry, respectively. The expressions of p-p38 MAPK, p-ERKs and p-JNKs in the myocardium were observed using immunohistochemical staining and Western blots.@*RESULTS@#No significant changes in cardiac function were detected before and after the perfusion in control group while UFPs perfused hearts showed a decline in cardiac function in a time-dependent manner (all P < 0.05). In UFPs-treated group, LVDP, +dp/dtmax, -dp/dtmax and CF were statistically reduced from (82.6±2.1) mmHg, (1 624±113) mmHg/s, (1 565±116) mmHg/s, (12.0±0.2) mL/min to (56.8±4.4) mmHg, (1 066±177) mmHg/s, (1 082±134) mmHg/s, (8.7±0.3) mL/min (all P < 0.05), respectively. Furthermore, The comparison between the two groups observed that UFPs perfusion caused a significant decrease in cardiac function at 30 and 40 min compared with the control group (all P < 0.05). At the end of the perfusion, the level of MDA was increased from (0.98±0.14) nmol/L to (1.95±0.18) nmol/L, while SOD and TAOC were reduced from (12.50±1.87) U/mL and (6.83±1.16) U/mL to (6.50 ±1.04) U/mL and (3.67±0.82) U/mL (all P < 0.001) in UFPs group, respectively. In coincidence with these changes, immunohistochemistry and Western blots results showed that the levels of p-p38 MAPK, p-ERKs and p-JNKs in the myocardium significantly increased in UFPs group as compared with control group (all P < 0.05).@*CONCLUSION@#The results of this study demonstrated that the short-term exposure of UFPs to the isolated rat hearts has direct and acute toxic effects on cardiac function, probably related to attenuation of anti-oxidative capacity and activation of MAPK signaling pathways.

Quality analysis of multi-source data for health management in major chronic diseases in Baoshan District of Shanghai / 上海预防医学

Objective An analysis of informationized multi-platform big data was conducted to learn about the quality change of health management data for hypertension and diabetes patients in Baoshan District of Shanghai since 2017.The result provided important information for further evaluation of the effect of quality control measures, and the prevention and management of chronic diseases. Methods Height, weight, blood glucose level, diagnosis and treatment information were collected from different databases of patients with hypertension and diabetes in Baoshan District from 2017 to 2019, and the consistency of the data from different sources was analyzed. Results Both the percentages of weight and height inconsistency among patients with hypertension and diabetes together were lower in 2019 than in 2017 (10.99% vs 18.72%, χ2=822.38, P < 0.001 and 0.86% vs 2.74%, χ2=347.03, P < 0.001, respectively).In 2019, the percentage of registered hypertensive patients with abnormal traceability from diagnosis was higher than that in 2017 (12.67% vs 11.72%, χ2=4.01, P=0.045).Similar results were also obtained in patients with diabetes.Analysis of glycated hemoglobin value last position in diabetic patients showed that the coefficient of variation of the last position composition ratio of the value in 2019 was significantly lower than that in 2017 (0.19 vs 0.31).The ratio in patients with the last position of glycosylated hemoglobin value of 0 was lower in 2019(4 701 cases, 12.72%) than that in 2017 (9 485 cases, 17.14%), and the difference was statistically significant. Conclusion The study result shows an improvement in quality management of hypertension and diabetes in Baoshan District of Shanghai.Information technology should be more widely used in promoting technical standardization, strengthening technical training, data quality control and effect evaluation.

Hepatoprotective Effect of Portulacae Herba on Carbon Tetrachloride Induced Acute Liver Injury in Mice / 中国实验方剂学杂志

Objective::To explore the effect and mechanism of Portulacae Herba protecting carbon tetrachloride (CCl4)-induced acute liver injury. Method::Sixty Kunming mice were randomly divided into normal group, model group, silybin group (200 mg·kg-1) and Portulacae Herba high, medium, low (2, 1, 0.5 g·kg-1) dose groups. After continuous intragastric administration for 5 days, mice in each group were intraperitoneally injected with 0.2% CCl4 peanut oil solution to establish acute liver injury model, except normal mice. After 23 hours of modeling, serum and liver tissue were collected. Fully automatic analysis of serum serum liver function indicators in mice. Liver tissues were taken for hematoxylin-eosin staining (HE) staining to observe liver pathological changes. RNA Sequencing (RNA-seq) was used to analyze differential genes and functional enrichment, real-time fluorescence quantification PCR(Real-time PCR) was used to verify the mRNA expression of cytochrome P450 family members(CYP)26A1, CYP2C37, CYP2C44, CYP2C50, CYP2C54. Result::Compared with normal group, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), total bilirubin (TBIL), malondialdehyde (MDA) in model group were significantly increased (P<0.05), and the activities of triglyceride (TG) and superoxide dismutase (SOD) were significantly decreased (P<0.05). Compared with model group, Portulaca Herba significantly reduced ALT, AST, TBIL and MDA levels in mice with acute liver injury (P<0.05), significantly increased SOD activity (P<0.01), and decreased the degree of liver tissue damage in mice. Compared with normal group, the mRNA expressions of CYP2C44, CYP2C50 in mice with acute liver injury were significantly decreased (P<0.05). Compared with model group, the mRNA expressions of CYP26A1, CYP2C37, CYP2C44, CYP2C50 and CYP2C54 were significantly increased in all dose groups of Portulaca Herba (P<0.05, P<0.01). Conclusion::Portulacae Herba has significant protective effects on acute liver injury caused by CCl4, and its mechanism may be related to the regulation of cytochrome P450 related genes.

Polyphyllin I alleviates osteoblasts injuries induced by TCP wear particles in vitro via inhibition of autophagy / 中草药

Objective: To observe the effect of polyphyllin I (PPI) on osteoblasts injuries induced by tricalcium phosphate (TCP) wear particles in vitro, and explain its regulation mechanisms. Methods: Primary osteoblasts obtained from the calvaria of neonatal SD rat by the series of digestion were identified with ALP staining. The osteoblasts were treated with TCP wear particles (TCP, 0.1 mg/mL) for 48 h to establish an in vitro injuries model of the calvarial osteoblasts. The experiment was randomly divided into control group, model (TCP, 0.1 mg/mL) group, PPI (30 μg/mL) group and PPI (100 μg/mL) group. CCK-8 and chemical colorimetry were used to examine cell viability and lactic dehydrogenase (LDH) content in culture media; Real-time PCR was performed to detect mRNA levels of ALP, Collagen I and RUNX2 in osteoblasts; The flow cytometry was used to examine apoptosis of osteoblasts using Annexin V/PI double staining; When the osteoblasts were treated for 14 d, mineral nodules formation was observed with alizarin S staining; Western blot was applied to examine proteins expression of Bax, Bcl-2, cleaved Caspase-3, Atg5, p62, and microtubule associated protein 1 light chain3 (LC-3). Results: Compared with control group, model group showed that the cell viability of osteoblasts, mRNA levels of ALP, Collagen I and RUNX2, and mineral nodules formation were significantly decreased; LDH content, percentage of apoptosis and proteins expression of Bax, cleaved Caspase-3, Atg5, LC-3 and the ratio of LC-3II/LC-3I were obviously increased in calvarial osteoblasts, whereas proteins expression of Bcl-2 and p62 was remarkably decreased. Compared with model group, PPI groups indicated that cell viability of osteoblasts, mRNA levels of ALP, Collagen I and RUNX2, and mineral nodules formation were dramatically increased; LDH content, percentage of apoptosis, protein expressions of Bax, cleaved Caspase-3, Atg5, and LC-3 and the ratio of LC-3II/LC-3I were obviously decreased, but Bcl-2 and p62 expression were obviously increased. Conclusion: PPI alleviates osteoblasts injuries induced by TCP wear particles via inhibition of autophagy.

Participation of Clinical Pharmacists in Pharmaceutical Care for a Child with Subglottic Talaromyces marneffei Infection / 中国药房

OBJECTIVE：To investigate the role of clinical pharmacists on the individualized treatment of children with subglottic Talaromyces marneffei infection. METHODS ：The clinical pharmacists participated in the medication procedure for a case of subglottic T. marneffei infection child . The clinical pharmacists suggested that Budesonide suspension for inhalation should be stopped，according to the subglottic infection pathogen type （T. marneffei ）；Itraconazole oral solution should be chosen and taken orally 2.5 mg/kg，q12 h，and indicators as liver function ，blood potassium should be monitored regularly. However ，as Itraconazole oral solution needed to be applied for temporary purchase ，Itraconazole capsules 2.5 mg/kg，q12 h，p.o.，was administrated temporarily ；clinical pharmacists suggested that Itraconazole capsules should be taken after meal ，and the doctor changed the feeding mode of milk from q 4 h to continuous pumping. After purchased ，Itraconazole oral solution was used instead 2.5 mg/kg，q12 h in fasting state ，and according the clinical pharmacist ’s suggestion ，the doctor changed the nursing method to q 4 h milk pumping. After purchasing and using oral solution instead ，clinical pharmacists suggested taking it at fasting state ；according to the monitoring results and target range （0.5-1 mg/L），oral dose of Itraconazole oral solution was finally adjusted to 8.3 mg/kg， q12 h. In view of the diarrhea during the treatment ，clinical pharmacists suggested to continue the original treatment after considering the effectiveness and importance of the treatment ；at the same time ，discharge medication education should be carried out. RESULTS ： The doctors adopted the suggestions of the clinical pharmacists. The child got a clinical improvement and was discharged after 48 days. CONCLUSIONS ：Clinical pharmacists participate in the treatment of children with T. marneffei infection，timely assist physicians to adjust and improve the medication regimen ，which improve the efficacy and safety of medication for children.

Therapeutic effects of different drug regimens and metabolic markers in children with bipolar affective disorder: a comparative analysis / 中国当代儿科杂志

OBJECTIVE@#To study the changes in metabolic markers and clinical outcome after treatment with different drug regimens in children with bipolar affective disorder.@*METHODS@#A retrospective analysis was performed on the medical data of 220 children with bipolar affective disorder who attended the hospital from January 2017 to January 2020. According to the treatment method, 112 children treated with atypical antipsychotic drugs alone were enrolled as the control group, and 108 children treated with atypical antipsychotic drugs combined with mood stabilizer were enrolled as the study group. The two groups were compared in terms of baseline data, changes in related metabolic markers[fasting insulin (FIN), glycosylated hemoglobin (HbAlc), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C)] after treatment, incidence rate of metabolic syndrome, and clinical outcome.@*RESULTS@#There were no significant differences in the baseline data including age, sex, and course of disease between the two groups (@*CONCLUSIONS@#Atypical antipsychotic drugs combined with mood stabilizer in the treatment of bipolar disorder in children have little effect on the level of metabolic markers, and the curative effect is significant.

Organ function support in patients with coronavirus disease 2019: Tongji experience / 医学前沿

Coronavirus disease 2019 (COVID-19) is a highly contagious disease and a serious threat to human health. COVID-19 can cause multiple organ dysfunction, such as respiratory and circulatory failure, liver and kidney injury, disseminated intravascular coagulation, and thromboembolism, and even death. The World Health Organization reports that the mortality rate of severe-type COVID-19 is over 50%. Currently, the number of severe cases worldwide has increased rapidly, but the experience in the treatment of infected patients is still limited. Given the lack of specific antiviral drugs, multi-organ function support treatment is important for patients with COVID-19. To improve the cure rate and reduce the mortality of patients with severe- and critical-type COVID-19, this paper summarizes the experience of organ function support in patients with severe- and critical-type COVID-19 in Optical Valley Branch of Tongji Hospital, Wuhan, China. This paper systematically summarizes the procedures of functional support therapies for multiple organs and systems, including respiratory, circulatory, renal, hepatic, and hematological systems, among patients with severe- and critical-type COVID-19. This paper provides a clinical reference and a new strategy for the optimal treatment of COVID-19 worldwide.

Effect of psoralen on rat osteoblasts injuries induced by TCP wear particles in vitro and its mechanism / 中国应用生理学杂志

To investigate the effect and mechanism of psoralen on calvarial osteoblasts injuries caused by tricalcium phosphate (TCP) wear particles in vitro. Primary osteoblasts were obtained from the calvaria of neonatal SD rat by the series of digestion and were identified with ALP staining. Calvarial osteoblasts were treated with TCP wear particles for 48 h to establish the in vitro model of osteoblasts injuries. The rat osteoblasts were randomly divided into control group, TCP wear particles (0.1 mg/ml) group, psoralen treated (at the concentrations of 10, 10, 10 mol/L) groups. WST assay and the flow cytometry were used to detect the cell viability of osteoblasts and apoptosis, respectively. Chemical colorimetry was performed to examine ALP activity of osteobalsts. When the osteoblasts were treated for 14 day, mineral nodules formation was observed with alizarin red S staining. Western blot was applied to examine protein expressions of glucose regulated protein78/94(GRP78/94), inositol dependent enzyme 1 alpha (IREα), spliced X-box binding protein 1 (XBP1s) and phosphorylated c-Jun N-terminal kinase (p-JNK) in calvarial osteoblasts. Compared with control group, the cell viability of osteoblasts, ALP activity and mineral nodules formation in TCP group were decreased significantly (P＜0.05), while the percentage of apoptosis and protein expressions of GRP78/94, IRE1α, XBP1 and p-JNK were obviously increased in calvarial osteoblasts (P＜0.05). Compared with TCP group, the injuries of calvarial osteoblasts and cell apoptosis in psoralen treated groups were obviously decreased (P＜0.05), and the expression levels of GRP78/94, IRE1α, XBP1 and p-JNK were down-regulated remarkably (P＜0.05). Psoralen prevents osteoblasts injuries caused by TCP wear particles through IRE1α-XBP1s-JNK signaling pathway activation.

Application of objective structured clinical examination in the training of professional postgraduate students in obstetrics and gynecology / 中华医学教育探索杂志

Objective To design and conduct objective structured clinical examination (OSCE) in professional postgraduates in obstetrics and gynecology, and to investigate the improvement measures for the teaching of professional postgraduates in obstetrics and gynecology by analyzing the performance of postgraduates in OSCE. Methods In October 2017, OSCE was performed for postgraduates in obstetrics and gynecology in the classes of 2016 and 2017 to evaluate their clinical ability. This OSCE consisted of the following four aspects: inquiry of standardized patients, basic practical skills in obstetrics and gynecology, utilization and interpretation of auxiliary examinations, and case analysis. The postgraduates were scored according to the following six items: doctor-patient communication skills, completeness of inquiry, basic practical skills in obstetrics and gynecology, utilization of auxiliary examinations, competence of clinical thinking, and mastery of medical knowledge. SPSS 18.0 was used for statistical analysis, and the t-test was used for comparison between groups . Results A total of 27 postgraduates participated in this OSCE . Among the six scoring items, completeness of inquiry had the lowest score (78 ±6), and utilization of auxiliary examinations had the highest score (85±3). Among the 27 postgraduates, 7 (25.9％) lacked doctor-patient communication skills, 6 (22.2％) showed serious problems in basic practical skills in obstetrics and gynecology, and 6 (22.2％) lacked the scientific thinking of clinical diagnosis. Conclusion In the training of professional postgraduate students in obstetrics and gynecology, doctor-patient communication skills and clinical thinking should be emphasized, and basic clinical skill training should be strengthened.

Study on correlation of thyroid hormone level, lipid and glucose metabolism and blood uric acid in patients with hyperthyroidism / 国际检验医学杂志

Objective To explore the correlation of thyroid hormone level, blood glucose level, blood lipid level and blood uric acid in patients with hyperthyroidism.Methods Totally 87patients with hyperthyroidism who were treated in our hospital from June 2015to December 2017were selected as the observation group, the other 69healthy people in the same period were selected as the control group.The fasting blood glucose (FBG), total cholesterol (TC), three glycerol (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), free three triiodothyronine (FT3), free thyroxine (FT4), thyroid stimulating hormone (TSH) and blood uric acid level were compared, and the correlation among the above indicators were analyzed.Results The levels of FBG, FT3, FT4and blood uric acid before treatment in the observation group were higher than those of the control group, and the levels of TSH, TC, TG, HDL-C and LDL-C were lower than those of the control group, the differences were statistically significant (P＜0.05).After treatment, the levels of FBG, FT3, FT4and blood uric acid were significantly decreased in the observation group, while the levels of TSH, TC, TG, HDL-C and LDL-C increased significantly, the differences were statistically significant between before and after treatment (P＜0.05).The levels of FT3and FT4were positively correlated with blood uric acid and FBG level in hyperthyroidism patients, and negatively correlated with the levels of TC, TG, HDL-C and LDL-C.The level of TSH was positively correlated with the levels of TC, TG, HDL-C and LDL-C, and had a negative correlation with the levels of FBG and blood uric acid.Conclusion The thyroid hormone level is closely related to lipid and glucose metabolism and blood uric acid level in hyperthyroidism patients.When the thyroid hormone is corrected, the blood glucose and blood lipid indexes turn to normal.It suggests that the clinical treatment of hyperthyroidism should be taken as the main means to correct the abnormal thyroid hormone.

Modeling of influenza-like illness prediction based on Elman neural network / 预防医学

Objective @#To build a model for influenza-like illness（ILI）prediction based on Elman neural network and to provide evidence for early warning of influenza epidemic in Zhejiang Province. @*Methods @#The data of ILI from 11 sentinel hospitals,influenza pathogen detection,meteorological factors and air pollutants in Zhejiang Province from 2013 to 2014 were collected. Time-delay correlation analysis was conducted to select variables for modeling. Based on Elman neural network,data from the 14th week of 2013 to the 44th week of 2014 were used as a training set to establish the model and the data from 45th week to 52nd weeks of 2014 were used as a test set for the model performance. @*Results @#There were ILI reported every week during 2013 and 2014,with a total of 506 391. The percentage of ILI cases per week was（3.07 ± 0.73）%. Ten variables selected by time-delay correlation analysis were the weekly average values of atmospheric pressure（13 weeks in advance）,vapor pressure（11 weeks in advance）,temperature（9 weeks in advance）,SO2（5 weeks in advance）,NO2（5 weeks in advance）,CO（5 weeks in advance）,PM2.5（5 weeks in advance）,PM10（5 weeks in advance）,air quality index（5 weeks in advance）and positive rate of pathogen（1 weeks in advance）. Elman neural network（10-15-1-1）was selected as the optimal model,and the prediction performed well,with 10.58% as the mean error rate and 0.876 7 as the nonlinear correlation coefficient. @*Conclusion @#This study demonstrated that Elman neural network including variables of meteorological factors,air pollutants and the positive rate of pathogen performed well on the short-term prediction of ILI incidence.

Application of anterior subcutaneous Internal Pelvic Fixation in pelvic anterior ring fracture: a meta analysis / 中华骨科杂志

Objective@#The related literatures on supra acetabular spinal pedicle screws and a subcutaneous connecting rod (INFIX) in recent years was reviewed, and the overall prognosis was analyzed by meta analysis.@*Methods@#literatures were obtained by searching the PubMed, Google Scholar, and Wanfang Full-text Database according to the criteria of inclusion and exclusion. The two members independently screened the qualified literature, evaluated the quality of the literature and extracted the data information. The data information includes demographic and surgical information, main results and secondary results. A large number of data in the selected articles were analyzed and summarized by meta.@*Results@#A total of 20 studies were included, including 567 patients. The clinical results were evaluated by Majeed standard in 6 articles, of which 5 articles described the number of excellent and general prognosis, including 185 patients. The excellent and good prognosis rate was 91% (95%CI: 80%-103%) by meta analysis. The general prognosis rate was 9% (95%CI: 3%-14%). Among the prognostic complications, the incidence of lateral femoral cutaneous nerve damage and heterotopic ossification was higher, and the incidence of other complications was less than 5%. There were 13 articles about the complications of ectopic ossification, of which 1 did not mention the specific number of asymptomatic heterotopic ossification, and the remaining 12 included 424 cases, including 132 cases of ectopic ossification. The incidence of ectopic ossification was 26% (95%CI: 11%-40%). Among the 567 cases, obvious symptoms of lateral femoral cutaneous nerve injury were reported. The patients with lateral femoral cutaneous nerve injury accounted for 25% (95%CI: 17%-32%), and the temporary nerve damage was found in 124 cases. The incidence of temporary agitation after subgroup analysis was 21% (95%CI: 14%-28%). Five articles mentioned the manifestations of postoperative femoral nerve damage, including 10 patients, the incidence of femoral nerve damage was 3% (95%CI: -1%-6%). A total of 18 articles on surgical site-related infections were reported, including 534 patients, of which 15 were infected, with an infection rate of 3% (95%CI: 2%-4%). 19 articles clearly reported the failure of internal fixation after INIFX operation, including 32 patients who needed reoperation for different reasons. The failure of internal fixation was 3% (95%CI: 2%-4%). A total of 585 cases of fracture non-prognosis data were reported, of which 473 cases were cured and 3 cases had bone nonunion. The non-union rate of fracture was 3% (95%CI: 1%-5%).@*Conclusion@#After InFix pelvic surgery, the patient's clinical follow-up prognosis score and imaging findings showed that the postoperative recovery was good, and the new technique also avoided the inconvenience and postoperative complications caused by many traditional pelvic external fixation techniques. However, it was also found that the lateral femoral cutaneous nerve damage and the incidence of heterotopic ossification were also higher.

Effect and Mechanism of Modified Buzhong Yiqi Tang and Pursed-lips Breathing on Serum TNF-α，IL-8，IL-6，IL-1β and Cys-C in Patients at Stable Stage of COPD / 中国实验方剂学杂志

Objective: To observe the effect and mechanism of modified Buzhong Yiqi Tang and pursed-lips breathing (PLB) on serum tumor necrosis factor-alpha (TNF-α), interleukin-8 (IL-8), IL-6, IL-1β and serum cystatin C (Cys-C) in patients of chronic obstructive pulmonary disease (COPD) at the stable stage. Method: Totally 120 cases of COPD treated at Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine from September 2017 to March 2018 conformed to the inclusion criteria, and were randomly divided into treatment group (60 cases) and control group (60 cases). Cases of control group received Tiotropium (18 μg/time, qd). In addition to the therapy of control group, cases of treatment group were also given Buzhong Yiqi Tang and PLB. Both groups were treated for 6 months. Percent of forced expiratory volume in one second in predicted value (FEV1%), ratio of FEV1 to forced vital capacity (FEV1/FVC), six-minute walk distance (6MWD),TNF-α, IL-8, IL-6, IL-1β and Cys-C were compared between both groups. Result: After treatment, the clinical efficacy rate of observation group was 93.33%, which was evidently higher than 86.67%of control group (Pa,IL-8, IL-6,IL-1β, Cys-C but increases in FEV1%, FEV1/FVC (Pa, IL-8, IL-6, IL-1β, Cys-C but increases in FEV1%, FEV1/FVC (PConclusion: Buzhong Yiqi Tang and PLB has an anti-inflammatory effect on COPD by reducing levels of inflammation cytokines TNF-α, IL-8,IL-6, IL-1β and chronic inflammation markers serum Cys-C and inflammatory reaction, and alleviating airflow obstruction, promoting the levels of FEV1%, FEV1/FVC, pulmonary function control inflammatory factors, so as to improve pulmonary function and the quality of life in patients with COPD.

A complex chromosome translocation with male infertility of karyotype analysis and literature review / 北京大学学报(医学版)

One case of family chromosomal karyotype with complex chromosomal translocation and male infertility was reported. This case is a male, 30 years old, Han nationality, who did not receive contraception for 3 years after marriage. The phenotype and intelligence of the patients were normal, and there were no abnormalities in the external genitalia. No abnormalities were found in the prostate and spermatic vein. There was no history of parotitis or testicular trauma, no history of smoking, drinking history, denial of harmful substances and history of radioactive contact. There were no similar patients in the family, and the secondary sex was normal. The routine semen examination suggested that the active sperm was seldom seen. There were no obvious abnormalities in the serum endocrine examination of the patient. Cytogenetic examination: the patient's karyotype 46XY, t (10; 18; 21) (q22; p11.2; q11.2). There was no deletion in locus sY84, sY86, sY127, sY134, sY143, sY254 and sY255. His wife's examination showed no obvious abnormality, and her karyotype was normal. The parents of the patients were not close relatives. Their father's chromosome karyotype analysis was 46, XY, and Y chromosome microdeletion was normal. The chromosome karyotype of the parent was 46XX, t (10; 18; 21), and the parents of the patient also had a daughter, whose phenotype and intellectual development were normal, chromosome karyotype 46XX, t (10; 18; 21). In this case, the patient's balance translocation should be inherited by the mother. Because of the normal phenotype of the patient, there was no loss of genetic material, but the abnormal chromosomes might be passed to the offspring, and the proportion of the unbalanced gametes was very high. Through systematic review and review of the cases, it was concluded that the balanced translocation carriers only changed the relative position of the translocation segments on the chromosomes, retained the total number of the original genes, only changed the relative position of the genes on the chromosomes, and had no serious effect on the role of the gene and the development of the individual. The phenotype was normal. The patients were given symptomatic treatment to improve semen quality. It is recommended that pre-implantation genetic screening/diagnosis(PGS/PGD) be performed if necessary. It is to guide married men and women to choose the appropriate childbearing age, avoid unhealthy environmental contacts, and strengthen genetic screening before and after pregnancy, so as to achieve the goal of eugenics.