AIM: To develop a reliable primary cultured hepatocytes model in vitro for liver metastasis research. METHODS: Hepatocytes were isolated by a modification of the two-step collagenase perfusion method. The apoptosis and cell cycle of hepatocytes were measured with flow cytometry. The proliferation of hepatocytes was detected by SRB method. RESULTS: The viability and purity of hepatocytes were 90% and 95%,respectively. The result of flow cytometry analysis showed that there was little apoptosis in hepatocytes and most of hepatocytes were in G_0/G_1 phase. The proliferation and albumin-secreting function of hepatocyte cultured by low glucose DMEM and high glucose DMEM were higher than that of cultured by RPMI1640 during 1 to 6 day, but there was no significant different between low glucose DMEM group and high glucose DMEM group. CONCLUSION: Hepatocytes have higher purity and viability with the normal biological activity for about 6 days by this method and it may be a cell model for the study of liver metastasis in vitro.

Objective To evaluate monocyte chemotactic peptide-1 (MCP-1) excretion levels in urine of renal transplant recipients and study the relation between urinary MCP-1 levels and acute rejection.Methods Urinary MCP-1 levels were determined by avidin biotin complex-enzyme linked immunosorbent reaction (ABC-ELISA).Results The urinary MCP-1 levels were significantly higher in recipients with acute rejection than in clinically stable ones (P

Objective To determine the effect of primary realignment of posterior urethral injury associated with pelvic fracture on length and delayed operative treatment of ensuing urethral stricture.Methods A retrospective review was made on the clinical data of 64 patients with posterior urethral injury after pelvic fracture treated from January 2008 to January 2012.Of those patients,43 underwent primary endoscopic realignment (early realignment group) and 30 received primary suprapubic cystostomy (cystostomy group).All were evaluated postoperatively for the late stricture rate,stricture length,types of delayed repair,and operation frequency.Results Rate of stricture was 53％ (18/34) in early realignment group and 100％ (30/30) in cystostomy group,but all were corrected by delayed urethroplasty.Mean length of the stricture was (1.8±0.6) cm in early realignment group and (2.9±0.7)cm in cystostomy group(t=6.7,P＜0.05).Of the urethrostenosis patients in early realignment group,83％ (15/18) were successfully corrected with a simple endoscopic cold incision and 17％ (3/18) with open surgery.In contrast,only 60％ (18/30) in cystostomy group were successfully corrected by endoscopic cold incision.Patients in cystostomy group underwent (2.8 ± 0.5) procedures for cure compared with (1.6 ± 0.6) procedures in early realignment group (t =9.2,P＜0.05).Conclusion Primary endoscopic realignment for posterior urethral injury pelvic fracture offers the decrease in stricture incidence,stricture length,operation difficulty and operation frequency.

Objective To evaluate the efficacy and safety of mesenchymal stem cells(MSCs)in preventing early acute rejection after renal transplantation.Methods Eighty-eight primary cadaveric renal allograft recipients in our department were randomized into two groups treated with bone marrow MSCs (BMSCs group,n =43) or not (control group,n =45).Main immunosuppressive therapy regimen consisted of steroids,tacrolimus or cyclosporine and mycophenolate mofetil in all recipients.Estimated glomerular filtration rate (eGFR) of transplant kidney,incidence of acute reaction (AR),graft survival and incidence of adverse events were recorded within 24 months.Results In BMSCs group,the incidence of AR was 4.7 ％ and 9.3 ％ at 3rd month and 6th month respectively,significantly lower than 20.0 ％ and 26.7 ％ (P＜0.05) in the control group.The eGFR at day 7,14and 30 post-transplantation was significantly higher in the BMSCs group than in the control group (P＜0.01,P＜0.01,P＜0.05 respectively).The incidence of adverse events in the BMSCs group and the control group was 44.2 ％ (19/43) and 66.7 ％ (30/45,P ＜ 0.05) respectively and the rate of infection was 37.2 ％ (16/43) and 33.3 ％ ( 15/46,P ＞ 0.05) respectively within 24 months.Conclusion Induction therapy with autogenous BMSCs appeared to be more effective in the prevention of AR following cadaveric kidney transplantation and was associated with better clinical outcomes as far as early renal graft function without compromising patient safety.

Objective To investigate the prevalence of iron deficiency(ID)and iron deficiency anemia (IDA) in pregnant women in urban areas of China. Methods The study was a national cross-sectional survey conducted from September 19th, 2016 to November 20th, 2016. According to the classification of the National Bureau of Statistics, all survey sites were set up in 6 regions of the country. Pregnant women were continuously selected using multistage stratified sampling. A total of 12 403 pregnant women were collected and examined for serum ferritin and hemoglobin levels. Results The median serum ferritin level during pregnancy was 20.60 μg/L(11.78-36.98 μg/L), the hemoglobin level was(118±12)g/L. With the progress of pregnancy, the levels of serum ferritin and hemoglobin decreased gradually. The median serum ferritin levels in the first, second trimester and third trimester were 54.30 μg/L(34.48-94.01 μg/L), 28.60 μg/L(16.40-50.52 μg/L), and 16.70 μg/L(10.20-27.00 μg/L)respectively(P<0.01). The mean hemoglobin levels were(127 ± 10)g/L,(119 ± 11)g/L and(117 ± 11)g/L respectively(P<0.01). The prevalence of ID in urban pregnant women was 48.16%(5 973/12 403), and IDA prevalence was 13.87% (1 720/12 403). The prevalence of IDA in the first, second trimester and third trimester were 1.96% (20/1 019), 8.40%(293/3 487)and 17.82%(1 407/7 897), respectively(P<0.01). The prevalence of standardized ID and IDA were significantly different in various regions of China(P<0.01). The standardized prevalence of ID were relatively higher in East China and Northeast China, 57.37% and 53.41% respectively, while it was the lowest in Southwest China, 30.51%. The standardized prevalence of IDA in South Central, Northwest, and East China were relatively high, 21.30%, 16.97% and 17.53% respectively, and the standardized prevalence of IDA in Southwest China was the lowest, 5.44%,the differents in various regions were significant(all P<0.01). Conclusion The current phenomenon of ID and IDA in pregnant women is still very common,and nutrition and health care during pregnancy should be strengthened.

OBJECTIVE：To investigate the inhibitory effects of total alkaloids of Gelsemium elegans （TAG） on the proliferation and angiogenesis of human colon cancer cells. METHODS ：Human colon cancer cell line HT- 29 and HUVEC were cultured in vitro . After the intervention of low- ，medium-，high-dose TAG （40，80，120 μg/mL），the morphology of the two cells was observed by fluorescence inversion microscope. The survival rate of HT- 29 cells and HUVEC was detected by CCK- 8 assay. Flow cytometry was used to detect HT- 29 cell cycle. The migration rate ，invasion rate and tube number of HUVEC were observed by scratching test ，Transwell invasion experiment and tube formation experiment. RESULTS ：Compared with blank group ，HT-29 cells and HUVEC were decreased to different extents in TAG groups ；dead cells were observed ，and the survival rate of both decreased significantly （P＜0.05 or P＜0.01）. The proportion of HT- 29 cells at G 2/M phase in TAG groups as well as those at G 0/G1 phase in medium-dose group were increased significantly ；the proportion of HT- 29 cells at S phase in TAG groups as well as those at G 0/G1 phase in high-dose group were decreased significantly （P＜0.05 or P＜0.01）. Survival rate ，migration rate and invasion rate of HUVEC were decreased significantly in TAG groups ，and tube number was also decreased significantly at each time point during 4-24 h（P＜0.01）. CONCLUSIONS ：TAG have inhibitory effect on the proliferation of human colon cancer HT- 29 cells and HUVEC，can change HT- 29 cell cycle ，inhibit the migration ，invasion and tube formation of HUVEC.

OBJECTIVE To explore the antitumor effect and mechanism of total alkaloids of Gelsemium elegans （TA） and sempervirine （SPV） in vitro. METHODS The effects of low， medium and high concentrations of TA （50， 100， 200 μg/mL） and SPV （10， 30， 50 μmol/L） on the morphology of human hepatoma cells （HepG2， Bel-7402）， human lung cancer cells （A549） and human colon cancer cells （HCT-8） were observed， and the toxicity of TA and SPV to four tumor cells was monitored. The effects of TA and SPV on the contents of caspase-3 and caspase-9 in the supernatant of HCT-8 cells， the protein expressions of phosphorylated protein kinase B （p-Akt） （Thr308， Ser473）， B-cell lymphoma 2 （Bcl-2）， Bcl-2-related X protein （Bax）， survivin， C/EBP-homologous protein （CHOP）， immunoglobulin binding protein （Bip） and microtubule-associated protein 1 light chain 3Ⅱ （LC3Ⅱ） in HCT-8 cells were detected. RESULTS After the intervention of TA and SPV， the volume reduction and nuclear shrinkage were founded in four tumor cells； the cell activity decreased to varying degrees， among which TA and SPV had the best inhibitory effect on HCT-8 cells. After the intervention of TA and SPV， the contents of caspase-3 and caspase-9 in the supernatant of HCT-8 cells， the protein expressions of Bax， CHOP， Bip and LC3Ⅱ all increased to different degrees， while the protein expressions of p-Akt （Thr308， Ser473）， Bcl-2 and survivin in HCT-8 cells all decreased to different degrees. CONCLUSIONS TA and SPV have inhibitory effects on the above four tumor cells， and the inhibitory effect on HCT-8 cells is the best. The mechanism of their action on HCT-8 cells may be related to promoting apoptosis， activating endoplasmic reticulum stress and autophagy.