Objective To investigate the apoptosis of cell line HL-60 induced by matrine and its possible mechanism. Methods CCK-8 assay was used to observe the proliferation inhibitation of HL-60 cells treated with matrine. FCM was applied to detect apoptotic cells and mitoehondrial transmembrane potential.Spectrophotometry was used to detect the activity of Caspase-9. Results Matrine (0.25-2.0 mg/ml) could significantly inhibit proliferation of HL-60 cells (F=67.83, P ＜0.05). Ater 48 hours, the apoptosis rate increased obviously in dose dependence (t = 4.685, 6.300. 9.641, 6.786, P ＜0.05). Within 48 hours, the mitochondrial transmembrance potential of HL-60 cells treated with matrine (1.0 mg/ml) gradually decreased (F = 54.83, P ＜0.05), and the activation of Caspase-9 gradually decreased in time dependence (F = 72.31,P ＜0.05). Conclusion Matrine can induce apoptosis of HL-60 cells by reducing mitochondrial transmembrance potential and activating Caspase-9.

Objective To explore effective treatment methods and experience of management and organization in the early period of multiple patients with extensive burns combined with blast injury of the lungs. Methods To summary and analyze the climical date of 16 patients with extensive burns the combined with blast injury and experience of management and organization. Results One patient died of severe blast injury of lung 5 days after,one died of multi-organ dysfunction syndrome, the others recovered well and the burn, healed well. Organization and the course of resuscitating patients were effective, logistic support was strong and all patients were transported and transferred successfully. Conclusion Individualization fluid infusion based on exact clinical observations and increaseing the ratio of colloid fluid would help to prevent shock accompany burst injury of lung. Early and suitable mechanical ventilation mode to improve ventilation/gas exchange function were important measures to manage inhalation injuries. To plan directive salvage schemes for emergency and powerful logistic support,and transporting and transferring patients in time would help to increase treatment efficiency of multiple patients.

Objective To explore effective methods of organization and early treatment of mass casualties of burn-blast combined injury. Methods Early clinical data of a batch of 16 patients with burn-blast combined injury and experience of organization for early care were summarized and analyzed. Results The plan for emergency care of mass casualties was implemented in time and well organized, as a result all the wounded survived the shock stage. 10 wounded servicemen were safely transported to other hospitals. Cardiac and respiratory arrest occurred in 1 case during transportation by ambulance, but was successfully resuscitated. Conclusions Timely deployment of plan for emergency treatment, with the organization of a task force including the director of the hospital and clinical specialists are pivotal in the success of taking care of mass casualties. Correct understanding of the clinical characteristics of burn-blast combined injury, effective fluid resuscitation in early stage of injury, and measures to prevent multiple organ dysfunction syndrome were essential in the treatment.

BACKGROUND:It is hopeful that skeletal muscle satellite cells(SMSCs)can be served as seed cells for hematopoietic reconstitution.Angelica polysaccharides(APS)can not only promote hematopoietic stem cells and hematopoietic progenitor cells proliferation and differentiation,but also change the growth characteristics of SMSCs.OBJECTIVE:To investigate the effects of APS on the proliferation of mouse SMSCs in different culture environments.METHODS:SMSCs were procured by a modified method from new born mouse.The α-actin protein of the SMSCs was examined by immunohistochemistry at 5 days after culture.SMSCs were cultured and synchronized for 24 hours in the 96-well plate.After that,SMSCs were assigned into the blank control group,marrow stroma cell supernatant group,APS DMEM/F12 groups(contained 50,100,200,300,400 mg/L APS)and the marrow stroma cell conditioned medium(disposed by 50,100,200,300,400 mg/L APS in DMEM/F12).The proliferation of SMSCs was determined by MTT.RESULTS AND CONCLUSION:The α-actin was positive in the cultured SMSCs.MTT results demonstrated that,SMSCs showed a proliferative property in the marrow stroma cell conditioned medium groups.Additionally,the marrow stroma cell conditioned medium can effectively alter growth characteristics of SMSCs in a dose-dependent manner.

Objective To evaluate the curative effect of insulin glargine plus acarbose in the treatment of elderly type 2 diabetes and cerebral infarction.Methods Totally 84 elderly patients with type 2 diabetes and cerebral infarction were received insulin glargine and acarbose for 16 weeks.The levels of fasting plasma glucose (FPG),hemoglobin A1c (HbA1c),Postprandial glucose values of 2 h (2 hPG),Postprandial C peptide (PCP) and Fasting C peptide (FCP) before and after treatment were detected.Results Compared with pretreatment,the levels of FPG,HbA1c and 2 hPG after treatment were decreased to (6.87 1.46) mmol/L (t =1.658,P＜0.05),(6.48±1.12)％ (t=1.629,P＜0.05) and (8.34±2.15) mmol/L (t=2.037,P＜0.05),respectively,while PCP and FCP were increased to (3.82±0.22) μg/L and (3.52±0.36) μg/L (t=2.698,t=2.087,bothP＜0.05).Conclusions Combined insulin glargine with acarbose in the treatment of elderly type 2 diabetes and cerebral infarction can effectively reduce fasting and postprandial blood glucose,and improve the function of islet β cells.

Objective To investigate the influence of arsenic trioxide (AS2 O3 )in the vasculogenic mimicry (VM ) of HepG2 cells, and to preliminary clarify the possible mechanism of inhibition of AS2 O3 on the VM. Methods Themean inhibitory concentration (IC50 )of AS2 O3 72 h after treatment of HepG2 cells was calculated by CCK-8 assay.The HepG2 cells were cultured on 3-D Matrigel and randomly divided into control group, 1/2 IC50 AS2 O3 group and IC50 AS2 O3 group.IPP software was used to calculate the number,length and area of VM,and the expression levels of VM-related proteins VE-cadherin and MMP-2, apoptotic-related protein caspase-3 and proliferation-related protein PCNA were detected by Western blotting method.Results The IC50 of AS2 O3 was 10μmol·L-1 72 h after treatment of HepG2 cells.The number,length and area of VM in 1/2 IC50 and IC50 AS2 O3 groups were significantly lower than those in control group (P<0.01);the number,length and area of VM in IC50 AS2 O3 group were also lower than those in 1/2 IC50 AS2 O3 group (P<0.05).Compared with control group,the expression levels of VE-cadherin and MMP-2 in 1/2 IC50 and IC50 AS2 O3 groups were decreased (P<0.05),and the expression levels of caspase-3 and PCNA had no significant change (P>0.05).Conclusion AS2 O3 can inhibit the forming of VM of HepG2 cells,which indicated that its mechanism may be related to inhibiting the expressions of VE-cadherin and MMP-2 .

Objective To observe the effects of high sucrose,high saturated fatty acid and high unsaturated fatty acid(diets) on insulin resistance and endothelium-dependent vasodilation function.Methods Adult Wistar rats were divided into normal control(NC) group,high sucrose(HS) group and high saturated fatty acid(HSF) group,high unsaturated fatty acid(HUF) groups.Insulin sensitivity was tested by hyperinsulinemic-euglucemic clamp after 24 weeks.Acetylcholine-induced(or sodium nitroprussideinduced) relaxation of preconstricted isolated renal arteries was measured by Mulvany myograph.Results GIR was obviously lower in experimental groups than that in NC group.GIR was negatively correlated with triglyceride (TG),free fatty acid(FFA).Acetylcholine-induced relaxation was markedly decreased in all experimental groups compared with that in NC group and the maximal response was decreased 37.4% in HSF group,32.7% in HUF group,27.7% in HS group.Acetylcholine-induced relaxation was enhanced by incubation with L-Arg and decreased incubated with L-NNA,MB in all experimental groups.Vasodilation response was negatively correlated with TG,INS and well positively correlated with NO,GIR.There was significantly negative correlation between FFA andNO.Conclusions: The rats fed high sucrose,high saturated fatty acid and high unsaturated fatty acid diets developed insulin resistance with reduced endothelium-dependent vasodilation function.

Objective To study the inhibitory effect and mechanism of dipeptide peptidase inhibitors analogues on LPS -induced inflam-matory response on microglia .Methods Microglia cells were cultured ,isolated and purified from the neonatal Sprague-Dawley rats.Divided them into blank group ,negative control ,LPS group and medicine group ( parallel determination for 3 times each group ) after pharmacological preconditioning for 48 hours.The optimal concentration of microglia proliferation induced by LPS were measured by MTT assay .Observed the role of dipeptide peptidase inhibitors analogues on LPS-induced microglia in different concentrations .The interleukin1β( IL-1β) ,tumor necro-sis factor alpha ( TNF-α) were assayed by enzyme-linked immunorrbent assay ( ELISA ) .The expression of TLR-4 was detected by Western blotting and the expression of NF-κB was detected by RT-PCR.Results LPS induced the proliferation of microglia and significantly in-creased the release of inflammatory cytokines in LPS-stimulated primary microglia .Compared with the blank group ,dipeptide peptidase inhibi-tors analogues could inhibit this effect and the IC 50 values was 1.014 ×10 -2 mol/L to MG after pretreatment for 48 hours.Dipeptide peptidase inhibitors analogues could inhibit the release of TNF-αand IL-1 significantly(P<0.01),and it decreased the expression of TLR4 and NF-κB signif-icantly(P<0.01).Conclusion This research suggests that dipeptide peptidase inhibitors analogues restrain cell proliferation and inflammatory re-sponse of LPS-stimulated microglia,and the possible mechanism may be related to the inhibition of the expression of TLR-4 and NF-κB.

Objective To investigate the prognostic value of plasma thrombomodulin(TM)in patients with septic shock.Methods A retrospective analysis was conducted on the clinical data of 180 patients with septic shock admitted to the intensive care unit of the 908th Hospital from May 2018 to November 2022.The patients were divided into survival group(106 cases)and death group(74 ca-ses)based on the 30-day follow-up outcomes.Propensity score matching(PSM)was used to match 57 surviving patients with 57 de-ceased patients in a 1∶1 ratio,based on confounding factors such as age,gender,underlying diseases,primary infection site,laborato-ry results and disease severity scores.TM and other coagulation molecular markers were compared between the two groups,and logistic regression,receiver operating characteristic(ROC)curve,survival and correlation analyses were performed.Results After PSM,the TM levels in the death group(18.3[13.2,22.3]TU/mL)were significantly higher than those in the survival group(13.7[9.0,18.3]TU/mL)(P＜0.05).Multivariate logistic regression analysis showed that TM was an independent risk factor for 30-day mortality in the patients with septic shock(OR=1.137,95％CI:1.023-1.262,P＜0.005).ROC curve analysis revealed that the areas under the curve(AUCs)for predicting 30-day mortality were 0.665,0.627 and 0.600 for TM,Acute Physiology and Chronic Health EvaluationⅡ(APACHE Ⅱ)and Sequential Organ Failure Assessment(SOFA)scores,respectively.Kaplan-Meier survival analysis stratified by the optimal TM cut-off value(17.9 TU/mL)showed that the 30-day survival rate of the TM＜17.9 TU/mL group was 1.56 times that of the TM≥17.9 TU/mL group(Log-Rank test,P＜0.000 1).Spearman correlation analysis demonstrated that TM levels were positively correlated with APACHE Ⅱ(r=0.10,P＜0.005)and SOFA scores(r=0.35,P＜0.005).Conclusion Plasma TM has showed a good predictive value for assessing the prognosis of patients with septic shock and may serve as a potential biomarker for determining the prognosis of septic shock.

It is one of the frequently utilized strategies for positive-negative selection to elevate the gene targeting efficiency in somatic cells by enriching targeted colonies. Knocking out prnp in animals by gene targeting can prevent it from expressing Prion protein (Pathogenic protein of transmissible spongiform encephalopathy), which enables it to resist infection of Prion. We constructed a bovine prnp biallelic targeting vector via the positive-negative selection strategy, and transfected the linearized vector into the bovine fetal fibroblasts through electroporation. Then, we selected cells in cell culture medium with G418 under a concentration of 600 microg/mL followed by Ganciclovir (GCV) under a concentration of 200 nmol/mL. In the end, we successfully obtained 176 cell clones. All these clones were identified by means of sequencing, immunofluorescence and western blotting, respectively, confirming that there existed 9 positive cell clones. The results showed that the bovine prnp gene was successfully knocked out. Conclusively, we provide an effective way to knockout bovine prnp gene, which could serve as the basis for producing prion protein gene knockout transgenic cloned cattle.
Animals ; Cattle ; Electroporation ; Encephalopathy, Bovine Spongiform ; genetics ; Fetus ; cytology ; Fibroblasts ; cytology ; metabolism ; Gene Knockout Techniques ; methods ; Gene Targeting ; Genetic Vectors ; genetics ; Prions ; genetics ; Transfection