Objective To investigate the effects of celastrol on the cell growth and apoptosis of human gallbladder cancer NOZ cells,and explore its potential molecular mechanism.Methods NOZ cell were cultured in vitro.And CCK-8 assay,Annexin V-FITC/PI staining method,cell cycle analysis were conducted to investigate the effects of celastrol on the growth and apoptosis of NOZ cells after being treated with drugs.The mitochondrial membrane potential and Bax and Bcl-2 protein expression level were determined by Rhodamine 123 and Western blot,respectively.Results Celastrol could inhibit NOZ cell growth,and the IC50 value was 5.3 μmol/L.Annexin-V/PI staining showed that cell apoptosis of NOZ cells were induced as the celastrol concentration increased,and the apoptosis ratio of control group was 4.4％,while the apoptosis rates of the test groups (2,5,10 p mol/L) were 7.4％,27.1％ and 43.4％,respectively.In addition,cell cycle analysis revealed that celastrol could induce G1-phase arrest.The G1-phase rate of control group was 25.6％,while the G1-phase rates of the test groups (2,5,10 μmol/L) were 36.5％,45.7％ and 92.5％,respectively.The mitochondrial membrane potential was measured after treatment with celastrol and the results indicated that the mitochondrial membrane potential was significantly decreased.Western Blot showed that the protein expression of Bax increased and Bcl-2 decreased in a time-dependent manner after treatment with celastrol.Conclusions Celastrol may inhibit cell proliferation of human gallbladder cancer NOZ cells and induce cell apoptosis partly by inducing the loss of mitochondrial membrane potential.

The paper is aimed to identify SNP in Sarcandra glabra and Chloranthus spicatus, and authenticate S. glabra from Ch. spicatus and the mixture by using PCR amplification of specific alleles. SNPs in the ITS sequences of S. glabra and Ch. spicatus were found by ClustulX 2. 1 program and Bioedit software. Primers for authentic S. glabra and Ch. spicatus was designed according to the SNP site, and ITS sequence universal primers plus to the authentic primer to construct a multi-PCR reaction system, and then optimized the PCR reaction system. Five hundred and eighty band special for S. glabra and 470 bp band special for Ch. spicatus were found by using multi-PCR reaction. The multi-PCR reaction system could be applied to identify S. glabra and Ch. spicatus's leaves.
DNA, Plant ; analysis ; genetics ; DNA, Ribosomal ; genetics ; DNA, Ribosomal Spacer ; analysis ; genetics ; Magnoliopsida ; classification ; genetics ; Plant Leaves ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; RNA, Ribosomal ; genetics ; RNA, Ribosomal, 18S ; genetics ; RNA, Ribosomal, 5.8S ; genetics ; Species Specificity

The study is aimed to assess the genetic diversity and genetic relationship of 18 Sarcandra glabra resources from different populations,and guide parent selection of cross breeding between these resources. The molecular marker technique ISSR was used to investigate the genetic diversity of the 18 resources. Data was analyzed by POPGEN 32, and a cluster diagram was presented by UPGMA. One hundred and ninety-eight amplified fragments were obtained using 23 ISSR primers. One hundred and eighty-four polymorphic loci were identified. Nei's genetic diversity index (h) was 0.32, Shannon diversity index (I) was 0.485 4. The genetic similarity coefficient among the resources ranged from 0.383 8 to 0.878 8 in an average of 0.661 2. The genetic distance between sample S2 and sample S18 was the farthest, so as between sample S3 and sample S18 both Nei's genetic distance was 0.957 5, The genetic distance between sample S4 and sample S5 was the closest, the Nei's genetic distance was 0.129 2,and the sample S1, S2, S3, S7, S10 were significantly different from the others based on the clustering analysis, the three groups S2 vs S3, S2 vs S6, S2 vs S18 were the best parent group selection. There was a middle level of genetic differentiation in the resources. The genetic distance between resources gives useful information to guide parent selection of cross breeding.
Conservation of Natural Resources ; DNA Primers ; genetics ; Genetic Variation ; Magnoliopsida ; classification ; genetics ; Microsatellite Repeats ; Phylogeny

AIM: To explore the serum levels of visfatin (VF) and tumor necrosis factor-alpha (TNF-α) in the patients with pre-eclampsia (PE) and their correlation with insulin resistance (IR).METHODS: The severe PE pa-tients (n =30), mild PE patients (n =30) and normal pregnant women (n =40) were selected according to the classifica-tion standard of PE.The serum levels of VF and TNF-αwere measured by ELISA.Fasting plasma glucose (FPG) and fasting insulin (FIns) were detected by glucose oxidase method and radioimmunoassay, respectively.Triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol ( HDL-C) and low-density lipoprotein cholesterol ( LDL-C) were measured by an automatic biochemical analyzer.According to calculating the mean arterial pressure (MAP), body mass index (BMI) and homeostatic model assessment for insulin resistance index (HOMA-IR), the correlation between IR and the levels of serum VF as well as TNF-αwere analyzed.RESULTS: The levels of VF and TNF-αin severe PE group and mild PE group were significantly lower than those in normal pregnancy group (P <0.05).In addition, the levels of VF and TNF-αin severe PE group were lower than those in mild PE group (P <0.05).Linear correlation analysis showed that serum VF was positively correlated with TNF-αand HDL-C (P <0.05), and negatively with MAP and FIns (P <0.05). The serum TNF-αwas positively correlated with HDL-C (P <0.05), and negatively with BMI, TG, MAP and FIns (P <0.05).Multiple stepwise regression analysis showed that FBG, FIns and HOMA-IR were relative independent factors of se-
rum VF and TNF-α(P <0.05).CONCLUSION: Serum levels of VF and TNF-αare closely related to IR.

The aim of this paper is to apply near infrared spectroscopy techniques to construct a rapid identification method for 8 kinds of mineral Chinese Medicines containing carbonates. The qualitative model using clustering analysis method in OPUS software can identify accurately 8 kinds of carbonate-containing mineral Chinese medicines. The near-infrared quantitative model was established by using partial least squares method (PLS) for 7 mineral Chinese Medicines in which main component is calcium carbonate. Compared with the results by EDTA titration, the established quantitative analysis model for calcium carbonate content showed a good prediction result that when the content is between 47.61% -99.17%, the average relative deviation of the prediction result is 0.24% and the average recovery rate was 100.3%. The results also showed that the model using near infrared spectroscopy can get not only a rapid identification of the 8 mineral Chinese medicines containing carbonates, but also an accurate and reliabe content determination of calcium carbonate for the 7 mineral Chinese medicines which contain the component.
Carbonates ; analysis ; Medicine, Chinese Traditional ; Minerals ; chemistry ; Software ; Spectrophotometry, Infrared ; methods ; Time Factors

Objective To investigate the efficacy of liver transplantation for the treatment of polycystic liver disease.Methods The clinical data of 8 patients with polycystic liver disease who received liver transplantation at the Third Affiliated Hospital from September 2003 to July 2009 were retrospectively analyzed.All patients underwent modified piggyback liver transplantation with vena cava plasty,and 1 of the patients received simultaneons liver-kidney transplantation.Data including acute rejections,complications,graft functions and survival time of patients were recorded.Results Operation was successfully performed on all patients.The mean operation time,anhepatic phase and operative blood loss were (7.5 +2.8)hours (range,4-11 hours),(42 + 14)minutes (range,25-70 minutes) and (2250 ± 1850)ml (range,2000-6500 ml),respectively.One patient received liver retransplantation at 45 days after primary liver transplantation because of hepatic artery stenosis.Two patients died during perioperative period.One patient died of pulmonary infection and multiple organ disfunction syndrome (MODS) at 39 days after operation,and the other one died of MODS at 59 days after operation.The median follow-up time was 60 months (range,37-93 months).Six patients survived for more than 3 years,4 patients survived for more than 5 years and 2 patients survived for more than 7 years.No acute rejection of gratis was observed at the end of the follow up.Conclusion Liver transplantation is safe and effective for the treatment of polycystic liver disease.

Objective To investigate the effects of recombinant human thioredox ( rhTrx) on primary cultured hepatocytes. Methods The cDNA coding hTrx was amplified by using RT-PCR. The objective protein was expressed in E. Coli. The activity of purified rhTrx protein was determined by IgM and insulin disulfide reduction assay. DNA synthesis was determined by 3H-TdR mixing assay to realize the proliferating roles of rhTrx in murine primary cultured hepatocytes. Cell viability was assessed by lactate dehydrogenase ( LDH) releasing assay. Results The cloning hTrx open reading frames cDNA was confirmed by sequence analysis. The soluble recombinant hTrx are expressed efficiently in the prokaryotic expression system. The recovery ratio was 23. 20% of total bacteria protein with a purity of the expressive protein of 96. 30%. The rhTrx had stronger IgM and insulin reduced activity compared with the controls (P

Objective To find the relevant factors of the young children’ physical and intellectual development in Guangdong province for children growth promotion.Method Physical status of 3 844 children 0-18 month and intellectual status of 749 children from urban,suburb and rural area in Guangdong province and their relevant factors were investigated.Then multiple regression analysis was applied statistically for their physical and intellectual development.Results Adjusted the interfering factors,the young children’ physical development was significantly correlated to their birth place,monthly income of family,monthly expense of food,paternal education degree,duration of breastfeeding,duration of exclusively breastfeeding,time of introduction of complementary food and intake of egg and vegetable.And their intellectual development was significantly correlated to their birth place,maternal education degree,time of introduction of complementary food and intake of formula,meat,bean and egg.Conclusion Increase of socioeconomic level,popularization of nutrition knowledge,promotion of breastfeeding and timely and reasonable introduction of complementary food are main effective measures to improve children’s development.

A strain no.H5 isolated from Rhizophora stylosa Griff in Zhanjiang had a good antagonistic activity against Aspergillus niger,pathogen of Dracaena Sanderiana black-rot disease.It was identified as Bacillus licheniformis.Dual culture,mycelium growth rate and inhibitory zones were used to test the effect. Strong inhibition was shown against A.niger.Inhibitory ratios of H5 germ-free fermented filtrate on mycelium growth and conidial germination were 91.9%and 100%respectively.In addition,mycelia on the edge of antagonistic band became abnormal and over-branching.Meanwhile,a lot of vesicles appeared on the surface.When treated with heat,acid and alkali,the filtration of H5 was always with stable activity.Precipitate in 55%saturated ammonium sulfate dissolved in phosphate buffer solution maintained most of the activity after high pressure steam sterilization for 25 minutes.It was preliminarily considered as a kind of heat resistant protein.

Objective To analyze the clinical characteristics, risk factors, prevention, and treatments of de novo malignant tumors after orthotopic liver transplantation (OLT). Methods The clinical data of 4 patients with de novo malignant tumors out of the 726 patients undergoing OLT from October 2003 to December 2008 were analyzed. Results The morbidity of de novo malignant tumors after OLT was 0.6 % (4/726), and all of them were men. The 4 de novo malignant tumors were respectively diagnosed as acute myeloid leukaemia, gastric carcinoma, lung cancer, undifferentiated embryonal sarcoma of the liver. The age of patients during OLT was 42-57 years (mean 52 years). The age of patients' onset was 45-60 years (mean 53 years). The length was 6-38 months from liver transplantation to diagnosis of de novo malignant tumors (mean 31 months). All of the 4 patients died of tumor progression and multiple organ failure. The survival time was 12-48 months after OLT (mean 39.5 months), and that was 6-10 months after diagnosis of de novo malignant tumors (mean 8.5 months). Conclusion The incidence of de novo malignant tumors after OLT in China is lower than in abroad. The delayed diagnosis time is a main cause of death for patients with de novo malignant tumors after OLT. To think highly of precancerous lesions and high risk of factors, early diagnosis, and early treatment are the keys to improve the survival time.