Objective To explore the role of hypoxia-inducible factor-1? (HIF-1?) in the expression of telomerase reverse transcriptase (TERT) in the pulmonary arterial smooth muscle cells (PASMCs) under anoxia.Methods HIF-1? and TERT mRNA in the PASMCs were examined by RT-PCR with the presence of HIF-1? inducers (CoCl_2,DFX),inhibitor (FAS,CBZ-LLL) and oligodeoxynucleotides,or under anoxia.HIF-1? protein in the PASMCs was determined by Western blotting.Results Expressions of HIF-1? and TERT mRNA significantly up-regulated under anoxia (0.59?0.07,0.60?0.06) compared with that in control group ( 0.11? 0.02,0.1?0.01,P0.05).Antisense oligodeoxynucleotides invalidated CoCl_2-induced increase in TERT mRNA levels (0.45?0.04) compared with CoCl_2 group (0.95?0.08,P

Objective To compare the clinical effect of two hemostatic methods for patients with pelvic fracture combined with hemorrhagic shock in the early stage.Methods A retrospective analysis was done on clinical data of 90 patients with unstable pelvic fracture combined with hemorrhagic shock managed by damage control resuscitation from January 2008 to December 2012.Unstable blood pressure and abdominal distension were noted postoperatively.Forty patients in Group A received laparotomy and bilateral internal iliac artery ligation from January 2008 to January 2010.Fifty patients in Group B underwent internal iliac artery embolization from February 2010 to December 2012.Comparative measurement was made on parameters of mortality,24-hour blood transfusion volume,24-hour lactic acid value,postoperative systolic blood pressure,postoperative body temperature,and postoperative prothrombin time (PT).Results Following parameters differed significantly between Groups A and B (P ＜ 0.05):mortality rate (53％ vs 12％),24-hour blood transfusion volume[(3 865.5 ±451.3)ml vs (2 108.8 ±336.4)ml],24-hour lactic acid value[(13.0 ± 2.0)mmol/L vs (5.4 ± 1.2)mmol/L],postoperative systolic blood pressure [(80.50 ± 22.73) mmHg vs (113.23 ± 20.89) mmHg],postoperative body temperature [(32.4 ± 0.2)℃ vs (36.1 ±0.3)℃],postoperative PT [(24.5 ±3.6)s vs (18.4±2.1)s].Conclusion For pelvic fracture combined with hemorrhagic shock,if the indications of abdominal viscera rupture are unclear,the interventional embolization can gain advantage over laparotomy in improving treatment success rate and reducing mortality and complications.

Objective To explore the protection and mechanism of hydrogen sulfide (H2S) preconditioning against injury induced by cisplatin in human marrow mesenchymal stem cells (HMMSCs).Methods HMMSCs were treated by cisplatin at 0,5,10,20,40,60,80 mg/L concentrations for 24 h respectively and were exposed to cisplatin at 20 mg/L concentrations for 0,6,12,24,36,48 h respectively.HMMSCs were pretreated with NaHS at 0,0.4,0.6,0.8,1.0 mmol/L respectively for 30 min before exposed to cisplatin at 20 mg/L concentrations for 24 h.HMMSCs were treated by U0126 or combined with human epidermal growth factor (HEGF) together for 30 min before they were preconditioned with NaHS for 30 min.The cell survival rate,cell apoptosis rate,the expression of p-ERK1/2 and t-ERK1/2 were recorded.Results The cell survival rate decreased to 71.72％±2.72％,59.41％±5.44％,50.37％±4.55％,38.97％±2.92％,30.11％±4.64％ and 21.71％±5.35％ respectively after cells were treated with cisplatin at 5,10,20,40,60,80 mg/L concentrations respectively,and the differences were statistically significant compared with 0 mg/L group.The cell viability fell to 70.30％±6.20％,61.63％±2.70％,51.29％±3.13％,38.72％±3.66％ and 27.57％±2.32％ after HMMSCs were treated with cisplatin at 20 mg/L for 6,12,24,36,48 h respectively,and the differences were statistically significant compared with 0 h group.The cell viability increased to 65.99％±2.67％,72.93％±5.44％,75.10％±4.71％ and 76.56％± 5.25％ when HMMSCs got pretreatment of NaHS,and the differences had statistical significance compared with cisplatin group.The cell apoptotic rate decreased from 35.29％±2.77％ to 18.62％±0.97％ when HMMSCs were pretreated with NaHS at 0.6 mmo/L.Treatment of HMMSCs with cisplatin at 20 mg/L for 24 h reduced p-ERK1/2 expression.The pretreatment of NaHS could inhibit the inhibitory action to the expression of p-ERK1/2 induced by cisplatin.Pretreatment with U0126 or HEGF inhibited or promoted the protection and the upregulated expression ofp-ERK1/2 caused by NaHS pretreatment.Conclusion The preconditioning of H2S can protect cell damage caused by cisplatin via activating the ERK1/2 pathway of HMMSCs.

Objective To investigate the effect of antisense telomerase reverse transcriptase (TERT) on 5-hydroxytryptamine (5-HT) induced proliferation and cell cycle of pulmonary artery smooth muscle cells (PASMCs). Methods PASMCs were cultured and divided into four groups: control group (cultured in RPMI-1640 culture medium), 5-HT group (cultured in culture medium containing 5-HT), antisense oligonucleotide (ASODN) group (cultured in culture medium containing 5-HT and ASODN TERT), and sense oligonucleotide (SODN) group (cultured in culture medium containing 5-HT and SODN TERT). The apoptosis of PASMC was observed by fluorescence microscopy with Hoechst staining. Apoptosis and cell cycle was analyzed with flow cytometry. Expression of proliferated cell nuclear antigen (PCNA) was determined by immunohistochemistry staining. Results Hoechst staining showed that apoptosis in 5-HT group (161?33) was significantly higher than that in control group (63?16, P

Objective To study the change characteristics of erythrocyte parameters in carriers of deletional alpha-thalassemia gene.Methods 389 patients with deletional alpha-thalassemia gene determined by the gap-PCR technique were classified into three groups based on different genotypes of alpha-thalassemia including silent thalassemia group,alpha-thalassemia trait group (265 ca-ses)and intermediate thalassemia group,and contemporaneous 188 healthy adults were randomly selected as the normal control-group(NC).The erythrocyte parameters including RBC,Hb,MCV,MCH,RDW were retrospectively analyzed and their differences were compared among aboved-mentioned groups by the analysis of variance and the multiple comparison.Results Alpha-thalasse-mia manifested by different degrees of microcyte hypochromia.There were statistically significant differences in the erythrocyte pa-rameters among various genotypes and phenotypes about erythrocyte indices(P <0.05).Moreover,the Hb,MCV and MCH values were lower than those in the control group,and had the decreasing tendency with the increase of deletedα-globin gene numbers;but the RDW value was higher than that in the control group,showing the increasing tendency,the differences had statistical siginifi-cance(P <0.05).Conclusion With the increase of deleted α-globin gene number,the characteristic of small-cell low-hemoglobin is more and more remarkable.RBC has a overall increase,while Hb,MCV and MCH have a decreasing tendency,and the heterogeneity of erythrocyte volume is increased.When MCV and MCH decreasing in high risk region,thalassemia should be highly suspected, but normal MCV and MCH can not exclude silent thalassemia and alpha-thalassemia trait.

Objective: To evaluate the differential calretinin immunostaining in different segments of total colonic aganglionosis and its utility in the diagnosis. Methods: Nine specimens including ileum and colon segments were obtained from 9 patients with total colonic aganglionosis (TCA), from 2010 to 2016 year, in Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science and Technology. Another 9 ganglionic specimens including the same segments from patients with non-Hirschsprung disease (non-HD) patients were collected as control. All cases were immunostained with calretinin. The patterns of calretinin immunostaining were observed, and morphometric analysis of each sample was performed by image analysis program (Image-Pro-Plus). The mean absorbance was evaluated by calculating the areas of the lamina propria occupied by the positively stained area of the calretinin at high power field. Results: The same pattern of calretinin immunostaining was seen in ganglionic ileum and ganglionic colon segments, with staining seen in intrinsic nerves fibers (INF), and in granular aggregates in the lamina propria and muscularis mucosae. There was no significant difference in the numbers of calretinin-positive INF from the ganglionic segments. In contrast, the number of calretinin-positive INF and granular aggregates in aganglionic segments were significantly lower than those in the ganglionic group (P<0.01). In the ileum transitional zone, scattered calretinin staining was observed, and the amount of calretinin-positive INF was significantly lower than those in the proximal segment of ganlionic ileum (P<0.01). Conclusions Since there is significant different expression of calretinin among the different segments from TCA, calretinin immunostaining has potential value in detecting TCA. It could be an important adjunctive method in detecting TCA in the future.

OBJECTI VE To optimize the extraction technology of the leaves of Dimocarpus longan according to flavonoids and phenolic acids. METHODS The contents of gallic acid ，protocatechuic acid ，ethyl gallate ，quercetin，luteolin and kaempferol in the leaves of D. longan were determined by HPLC. Based on single factor test ，with the ethanol volume fraction ，solid-liquid ratio and extraction time as factors ，using comprehensive scores of the contents of above six components as indexes ，the extraction technology of the leaves of D. longan was optimized by Box-Behnken response surface methodology. RESULTS The optimal extraction technology included ethanol volume fraction of 100％，solid-liquid ratio of l ∶ 7（g/mL），extraction time of 90 min， extraction temperature of 80 ℃. After 3 times of validation tests ，the average comprehensive score was 97.54（RSD＝0.33％，n＝ 3），relative error of which with predicted score （99.05）was 1.55％. CONCLUSIONS Box-Behnken response surface methodology combined with multi-index comprehensive score can be used for the extraction technology of the leaves of D. longan ，and the optimized extraction technology is stable and feasible.

Objective To investigate the active ingredients of Jingfang Baidu San for the prevention and treatment of COVID-19 by using network pharmacology and molecular docking, and to provide references for clinical applications. Methods The chemical constituents and action targets of all medicinal materials in Jingfang Baidu San were retrieved from TCMSP. Uniprot database was used to search the corresponding genes of targets. Cytoscape software was used to construct the network of medicinal materials-compounds-targets for visualization. The target proteins of COVID-19 were searched by disease databases. The intersection of both was considered to be analyzed to establish the protein-protein interaction (PPI) network by STRING database. GO function enrichment analysis and KEGG pathway enrichment analysis were performed through Metascape database to predict its mechanism. The effective strength of core constituents on preventing COVID-19 was calculated by molecular docking method. Results A total of 159 effective ingredients and 277 potential targets were obtained in Jingfang Baidu San within the given screening conditions [oral bioavailability (OB) ≥30%; drug-like (DL) ≥ 0.18], including 55 core targets with the intersection of 273 targets of COVID-19. According to the results of GO and KEGG enrichment analysis performed on the core targets, 1376 GO items and 136 KEGG pathways were obtained, involving infectious diseases, cancer, cell progress, immune system, signaling pathways etc. The results of molecular docking indicated strong binding capacity between the core ingredients and SARS-CoV-2 3CL hydrolase or angiotensin-converting enzyme II (ACE2). The hydrogen binding and hydrophobic effect were the main forms of the interaction. Conclusion The active ingredients in Jingfang Baidu San can inhibit the binding between SARS-CoV-2 protein and ACE2, thus regulating multiple targets and signal pathways, which plays a role in the prevention and the treatment of COVID-19.

OBJECTIVE To study the metabolites derived from the ethanol extract from the leaves of Dimocarpus longan preliminarily in rats in vivo ，and to provide reference for elucidating the possible metabolic mechanism of the leaves of D. longan in lowering blood glucose . METHODS Ultra high performance liquid chromatography quadrupole time -of-flight mass spectrometry （UPLC-Q-TOF-MS/MS） was adopted by taking ethanol extract of D. longan leaves，the feces and urine of rats at 0-72 h and 0-48 h after intragastric administration of 33.8 g/kg ethanol extract of D. longan leaves（by extract ），the feces and urine of rats at the corresponding time after intragastric administration of normal saline （blank control ） as samples . The accurate relative molecular weight ，formula and fragment information of the compounds were collected ， and the compounds were speculated and i dentified by matching with the database and spectrum library of the instrument ，and comparing with the reference substance and relevant literature . RESULTS A total of eight compounds were identified in urine and feces of rats ，including 2 prototype components and 6 metabolites. Three compounds （including two prototype components as quercetin ，luteolin and one metabolite as luteolin or kaempferol） in feces of rats were identified ；five compounds （all metabolites ） in urine of rats were identified ，involving metabolites of quercetin ，luteolin or kaempferol . Metabolites mainly included the products of methylation ，glucuronidation and oxidation. CONCLUSIONS After intragastric administration ，the ethanol extract from the leaves of D. longan is mainly metabolized in rats through methylation ，glucuronidation and other pathways . The identified compounds are mostly metabolites of quercetin and luteolin .