Due to the increasing use of hepatoeytes,the demand of hepatoeytes is increasing consistently in basic clinical research.The application of rational and efficient method of hepatoeyte isolation is a prereq-uisite for its application.At present,there are many methods of hepatoeyte isolation.This review summarizes the latest progress on them.

Objective To observe the protective effects of the mixture of ferulic acid (AF) and paeoniflorin on PC12 cells injury by FeSO4/H2O2 in vitro, and explorer its mechanism. Methods The cell injury model was established with FeSO4/H2O2 for 24 h. The process of the growth and morphology of the cells were observed under phase contrast microscope in vitro. The survival rate of the cells was determined by MTT method. The content of MDA and SOD activity was measured respectively by thiobarbituric acid and xanthine oxidase method. Results Compared with FeSO4/H2O2, the mixture of ferulic acid (AF) and paeoniflorin enhanced the PC12 cells survival rate and viability, reduced MDA content and increased SOD activity. Conclusion The mixture of 5～50 ?mol/L AF and 1～10 ?mol/L paeoniflorin exerted protective effects on PC12 cells injury induced by FeSO4/H2O2, which maybe related to free radical scavenging activity and lipid peroxidation process being inhibited.

Objective:To analyze the effect of insulin aspart on the islet cell secretion function, ultrasound imaging indicators and maternal and infant outcome in patients with gestational diabetes mellitus (GDM).Methods:The clinical data of 120 patients with GDM in Beijing Obstetrics and Gynecology Hospital Affiliated to Capital Medical University from February 2017 to July 2019 were retrospectively analyzed. Among them, 60 cases were treated with insulin aspart (observation group) and 60 cases were treated with biosynthetic human insulin (control group). The efficacy indexes, ultrasound imaging indexes and maternal and infant outcome were compared.Results:The 2 h postprandial blood glucose (2 h PBG) after breakfast on the third and fifth day of treatment in observation group was significantly lower than that in control group: (6.85 ± 0.87) mmol/L vs. (7.47 ± 1.35) mmol/L and (6.24 ± 0.59) mmol/L vs. (6.60 ± 0.87) mmol/L, and there was statistical difference ( P<0.01); there was no statistical difference in 2 h PBG after breakfast on the first day of treatment and after lunch and dinner on the first, third and fifth day of treatment between 2 groups ( P>0.05). The homeostasis model assessment-insulin resistance index (HOMA-IR) after treatment in observation group was significantly lower than that in control group (2.57 ± 0.25 vs. 3.00 ± 0.35), the homeostasis model assessment- β cell function index (HOMA-β) was significantly higher than that in control group (72.45 ± 12.33 vs. 63.66 ± 10.72), and there were statistical differences ( P<0.01). There were no statistical differences in pre-lunch blood glucose, pre-lunch initial insulin amount, pre-lunch final insulin amount and blood glucose target time between 2 groups ( P>0.05); the incidence of pre-meal hypoglycemia in observation group was significantly lower than that in control group: 8.33% (5/60) vs. 23.33% (14/60), and there was statistical difference ( P<0.05). The umbilical artery and renal artery resistance index (RI), maximum systolic blood flow velocity and end-diastolic blood flow velocity ratio (S/D) after treatment in observation group were significantly lower than those in control group (RI: 0.49 ± 0.16 vs. 0.59 ± 0.15 and 0.69 ± 0.17 vs. 0.76 ± 0.12; S/D: 2.09 ± 0.22 vs. 2.38 ± 0.26 and 5.17 ± 0.45 vs. 5.77 ± 0.63), and there were statistical differences ( P<0.01 or <0.05). There were no statistical differences in gestational age, delivery mode, neonatal body weight and the incidences of macrosomia, neonatal hypoglycemia and neonatal referral between 2 groups ( P>0.05). Conclusions:Insulin aspart can significantly improve the islet cell secretion function and ultrasound imaging indexes in the treatment of GDM, and can reduce the risk of pre-meal hypoglycemia, but the effect on maternal and infant outcome remains to be explored.

ObjectiveTo study the mechanism of infection-related reactive thrombocytosis.MethodsSeventy-five infectious disease patients were selected including 42 cases with acute infections phase thrombocytosis (acute infectious phase thrombocytosis group),18 cases with acute infectious phase normal platelet count (acute infectious phase normal platelet count group),15 cases with recovered phase (infectious recovered phase group) and 16 cases with healthy controls(control group).The serum thrombopoietin (TPO),intefleukin-6 (IL-6),white blood cell,platelet was determined and compared among 4 groups.Results The serum TPO in acute infectious phase thrombocytosis group [( 159.1 ± 65.9) ng/L]was higher than that in acute infectious phase normal platelet count group,infectious recovered phase group,and control group [(43.5 ± 14.4),(40.3 ± 15.2),(41.8 ± 18.9) ng/L](P＜ 0.05).There was no significant difference in the serum IL-6 between acute infectious phase thrombocytosis group [(542.7 ± 247.0) ng/L]and acute infectious phase normal platelet count group [(598.5 ± 250.4) ng/L] (P ＞ 0.05 ),but which was higher than that in infectious recovered phase group [(43.5 ± 20.7 ) ng/L] and control group [( 38.3 ± 17.6 )ng/L] respectively (P ＜ 0.05 ).The serum IL-6,TPO was positively correlated with platelet,white blood cell.The serum TPO was positively correlated with IL-6.ConclusionElevated TPO leads to the thrombocytosis,which is the possible mechanism of infection-related reactive thrombocytosis.

Objective To build of lentiviral mediated RNA interference of mammalian target of rapamcin (mTOR) ,because the mammalian target of rapamycin plays a important role in tumor development and the signal path .Methods According to RNA in-terference (RNAi) design rules Completely ,in view of the gene called mTOR was desiged four interference targets and Negative control (FAM ) sequence ,first of all ,synthetic oligonucleotides nucleotide fragments with artificial ,and can obtain siRNA fragments effectively by the method of PCR joining together ,then undertake transfection on lung adenocarcinoma A 549 cell by Lipofectamine transfection reagent 2000 .To begin to observe the enhanced green fluorescent protein expression in lung adenocarcinoma A 549 cells by fluorescence microscopy at high magnification after 1 days .We can use semi-quantitative RT-PCR method ,and detect of mTOR gene expression of mRNA level after 1 days ,meanwhile ,testing the expression of protein levels by Western Blot after 2 days ,in or-der to select the most efficient interference target sequence ,,afterwards ,synthetic double-stranded DNA ,and it can be make up vec-tor system with plasmid pHelper 1 .0 and pHelper 2 .0 by the pGCL-GFP carrier ,further transfect 293 T cells ,at last produce lenti-viral after packaging ,then detection GFP protein expression levels by Western-Blot method ,and consequently detect the virus drops degree of 293T cells ,at the same time ,identify the activity .Results The high efficiently target of mTOR gene has been successfully selected ;mTOR siRNA infecte of human lung adenocarcinoma A549 cells in vitro culture ,and the gene is obviously silence no mat-ter from the mRNA level or protein level ;3 mTOR gene lentivirus siRNA carrier was successfully build .geting virus supernatant al-so ,and virus drops to 1 × 108 UT/mL .Conclusion MTOR siRNA infected of human lung adenocarcinoma A549 cells in vitro cul-ture ,and could lead to mTOR gene obviously silence ;The construction was successfully gene mTOR siRNA lentivirus vectors .

Objective To investigate the relationship between body mass index (BMI) and long-term survival in patients with chronic obstructive pulmonary disease (COPD).Methods A retrospective cohort study was conducted in 1124 patients with completed data among 1528 hospitalized patients with COPD.Vital status was ascertained at death registry and civil affairs department.Clinical characteristics were acquired from medical record.Patients were divided into 4 groups according to the quartile of BMI:BMI＜18.9 kg/m2 (group A),BMI from 18.9 kg/m2to 22.7 kg/m2 (group B),BMI from 22.8 kg/m2 to 26.2 kg/m2 (group C),BMI＞26.2 kg/m2 (group D).The differences in survival curves were compared between groups by using log-rank test.The relationship between all-cause mortality and BMI was evaluated by Cox proportional hazards regression model.Results The median BMI of 1124 patients was 22.7 kg/m2 [(18.9-26.2) kg/m2].270 patients (24.0％) had BMI＜18.5 kg/m2.162 patients (14.4％) died during follow-up.There was significant difference in the survival curves in the four groups (x2 =7.97,P＜0.05).Survival rate was highest in group C and lowest in group A.After adjustment for other factors,BMI was an independent risk factor for predicting long-term survival.The survival rate was respectively decreased by 41 ％ and 35 ％ in group C and D as compared with group A.Conclusions Weight loss is comman in COPD patients.BMI is an independent risk factor for predicting long-term survival.BMI is easily acquired and stable,which is especially suitable to evaluate the prognosis of patients with acute exacerbation.

Objective To investigate the association between inflammatory biomarkers and survival in patients with stable chronic obstructive pulmonary disease(COPD).Methods A total of 1038 patients with stable COPD from January 2008 to December 2009 were included in a prospective cohort study.Clinical characteristics,pulmonary function tests,6 min walk test and a modified British Medical Research Council dyspnea scale (MMRC) were completed.Fasting blood was obtained to detect inflammatory biomarkers including neutrophils,C-reactive protein(CRP),fibrinogen,TNF-a,IL-6 and IL-8.Participants were evaluated every 3 months,all-cause mortality was used as the end event.Results 120 patients (9.2％) died in the period of follow-up.Kaplan-Meier analysis showed that 1 year,2-year-and 3-year survival rates were 94.4％,88.3％ and 84.0％,respectively.Compared with survivors,those who died had a higher level of inflammatory biomarkers.Cox multivariate regression analysis demonstrated that the independent risk factors for death were neutrophils (HR:1.262,95％CI:1.143-1.512,P=0.035),CRP (HR:1.234,95％CI:1.097-1.624,P=0.029),fibrinogen (HR:1.327,95％CI:1.141-1.619,P=0.026),TNF-α (HR:1.124,95％CI:1.043-1.659,P=0.045),IL-6 (HR:1,429,95％CI:1.237-1.816,P=0.014) and IL-8 (HR:1.188,95％CI:1.024-1.383,P=0.042).C statistical analysis showed that no single biomarker significantly improved the C statistic value on the base of clinical model,but it was further improved by the addition of all biomarkers (C =0.764,P =0.010).Conclusions The level of inflammatory biomarkers in the death with stable COPD is significantly increased.Age.BODE index,neutrophils,CRP,fibrinogen,TNF-α,IL-6 and IL-8 are independent risk factor for the prediction of mortality in patients with COPD.

Objective To investigate the risk factors of acute kidney injury(AKI) after intracoronary stent implantation in order to provide the basis for clinical prophylaxis and treatment.Methods Retrospectively analyzed 626 consecutive patients who underwent isolated intracoronary stent implantation in our institution from January 2007 to July 2011.Multivariate logistic regression model was constructed to identify the risk factors for the development of AKI defined as a serum creatinine (SCr) 130 to 199 μ mol/L or estimated creatinine clearance(Ccr) 30 to 60 ml/min per 1.73 m2.Results Ninety-three patients of 626 (14.9％) underwent isolated intracoronary stent implantation developed AKI.The results of the multivariate forward stepwise logistic regression analysis found that risk factors for the development of AKI following isolated intra-coronary stent implantation was associated with age (OR =1.570,95％ CI 1.308-1.885),ejection fraction (EF) ≤ 30％(OR =11.526,95％ CI 2.452-54.177),hypotension during perioperative and postoperation (OR =11.074,95％ CI 2.439-50.282),operation duration(OR =1.032,95％ CI 1.012-1.051),sex (OR =0.010,95％ CI 0.001-0.086),NYHA class Ⅲ & Ⅳ (OR =0.209,95％ CI 0.059-0.737),peripheral vascular disease (OR =0.528,95％ CI 0.286-0.973),chronic obstructive pulmonary diseases (OR =0.546,95％ CI 0.304-0.982),preoperation Cr (OR=1.418,95％CI 1.216-1.654) (and all P＜0.05).Conclusion AKI is the common complications after intracoronary stent implantation,especially age,EF ≤ 30％,hypotension during perioperative and postoperation,operation duration are independent risk factors.

Objective To investigate the impact of high intra-abdominal pressure on intracranial pressure in patients with traumatic brain injury and discuss the clinical significance of abdominal decom-pression. Methods Intra-abdominal pressure and intracranial pressure of 15 patients with abdominal trauma and brain injury were observed to discuss changes of intracranial pressure after abdominal decom-pression. Results After abdominal decompression, all patients got lower intracranial pressure, with decrease of (15.2±3.6) mm Hg. Conclusion Intra-abdominal pressure does affect intracranial pres-sure for patients with abdominal trauma and brain injury. Abdominal decompression may be effective for high intracranial pressure.

Objective To compare the effect of three cryoprotectants on vitrification-cryopreservation of C57BL/6J mouse epididymis.Method Epididymises from 6-8-week old male C57BL/6J mice (age) were cryopreserved using DMSO, PROH and R18S3 cryoprotectant solution and thawed , respectively.The morphology of sperm from thawed epididy-mis, the rate of post-thawing survival and reproductive capacity were determined to evaluate the freezing efficiency of the three cryoprotectants .Results The sperms from thawed epididymis of the three groups were all well-preserved structural-ly.The survival rate of sperms was 88.17%, 61.17% and 16.83% in the PROH, DMSO and R18S3 cryoprotectant solu-tions, respectively, and there were significant differences between the three cryopreservation groups (P<0.05 for all). The number of pups from the DMSO , PROH and R18S3 groups were 13, 8 and 17 mice after ICSI and embryo implantation , respectively.Conclusions All the three cryoprotectants DMSO , PROH and R18S3 solutions are suitable for vitrification-cryopreservation of C57BL/6J mouse epididymis.But PROH is the preference of these cryoprotectant solutions .