Objective To investigate the influence of coix seed triglyceride combined with transcatheter arterial chemoembolization (TACE) therapy on AFP, CD4﹢CD25﹢regulatory T (Treg) cells and cellular immune function in patients with advanced primary hepatocellular carcinoma (HCC). Methods A total of 50 patients with inoperable HCC, whose imaging examination showed no distant metastasis, were divided into the study group (n=25) and the control group (n=25). Coix seed triglyceride together with TACE was employed for the patients of the study group, while only TACE was adopted for the patients of the control group. For the patients of the study group, transcatheter hepatic artery infusion of 100 ml coix seed triglyceride was carried out during the performance of TACE, and postoperative intravenous drip of coix seed triglyceride (200 ml/d) was used for 5 days. The peripheral blood samples were collected one week before and one month after the treatment to detect the changes of AFP and T lymphocyte subsets (CD3﹢, CD4﹢, CD8﹢,CD4﹢/CD8﹢ and Treg) levels. One month after the treatment, enhanced CT, MRI or PET-CT was performed to evaluate the necrosis degree of the tumor. Results After the treatment, AFP levels was decreased in both groups, when compared the preoperative data the differences were statistically significant (P<0.01); the tumor necrosis rate of the study group was (57.7 ±8.2)%, which was slightly higher than (57.7±8.2)% of the control group, however, the difference was not statistically significant (P>0.05). In the study group, the percentage of Treg cells decreased from preoperative (8.27±6.65)%to postoperative (4.22± 1.59)%, the difference was statistically significant (P<0.01). The percentages of CD3﹢ and CD4﹢ and the ratio of CD4﹢/CD8﹢ increased from preoperative (55.78 ±13.66)%, (43.98 ±14.00)% and 1.22 ±0.64 to postoperative (62.29±10.78)%(P<0.01),(51.82±16.32)% (P<0.05) and 1.54±0.80 (P<0.05) respectively, while the percentage of CD8﹢decreased from preoperative (45.71±12.94)%to postoperative (39.70±12.41)%(P<0.05). In the control group, no statistically significant differences in the above mentioned indexes existed between preoperative data and postoperative ones (P>0.05). Conclusion In treating advanced primary HCC, coix seed triglyceride combined with TACE can reduce the percentage of Treg cells, thus, influence the patient’s cellular immune status and possibly decrease the recurrence rate of HCC after TACE therapy.

The Department of Obstetrics and Gynecology has performed series of teaching reforms for the seven-year medical students.For the theory classes,all the teachers are the senior ones who can enhance the bilingual teaching and combine the clinical cases with the latest development in the subject.We let the interns follow the diagnosis and treatment of the patients just as the patients'relatives.We also let a tutor to guide each intern to help him master the basic clinical art.During the ward inspection for teaching,we let the students be the main speaker,hoping that we can build a new student-centered teaching model.

To establish a HepG2 cell line ,which stably expressing CYP3A29 of Bama miniature pig in order to evaluate the drug metabolic characteristics of this isoenzyme,the gene for this system was obtained through total RNA extraction and RT-PCR assay.the gene was subcloned into plasmid PMD18-T,designated as pMD-CYP3A29.This gene was then amplified by PCR, and cloned into eukaryotic expression vector pcDNA3.1(+),and the recombinant plasmid was designated as pcDNA-CYP3A29. Sequencing was used to confirmed the correctness of the gene of this gene.the expressed gene was then transfected into HepG2 cells by lipid-media transfection and the transformants were screened by G418 for 10 generations ;the expression of CYP3A29 was identified by RT-PCR、West-blot and the metabolic activity of the transformant HepG2-CYP3A29 was verified by nifedipine oxidation.In comparison with HepG2, the transformant HepG2-CYP3A29 showed remarkable oxidative activity.It is apparent that the cell line stably expressing CYP3A29 isoenzyme was successfully established, and it may be used for the metabolic study of related drugs.

To gain a better understanding of the genetic diversity and evolution of PRRSV in the Ningxia Hui Nationality Autonomous Region (Ningxia) of China, the nsp2 genes from a series of PRRSV strains collected from the region in 2007 were partially sequenced. These sequences were then analyzed along with the classical strain (ch-la) and two other epidemic strains SD (3) and SD2006. Comparison of the nucleotide sequence with ch-la indicated that nsp2 genes of seventeen Ningxia isolates (NX strain) have deletions of 87 nucleotides. Sequence analysis indicated that homology between the Ningxia strain and ch-la was 60.3%-79.9% in the nucleotide sequence, and homology between the NX strains and SD strains was 80.3%-98.8% in the nucleotide sequence. The nsp2 genes of the seventeen isolates had 74.9%-100% nucleotide sequence identities with each other. This study was undertaken to assess the regional variation of prevalent PRRSV and to establish a sequence database for PRRSV molecular epidemiological studies.

The bacteriophage T7 RNAP gene was amplified via PCR from -lysogen DE3, and the gene was cloned into pBABEpuro retrovial vector, a recombinant plasmid named as pT7BABEpuro was constructed and sequenced. Then the pT7BABEpuro and pVSV-G plasmids were cotransfected into GP2-293 packaging cells by liposomese, some pseudotype viruses were ingathered and transfected into IBRS-2 cell under polybrene. The IBRS-2 cell was propagated in DMEM with puromycin. The genome extraction from the cells transfected different times, the T7 RNAP gene was amplified from the genome by PCR, the mRNA of T7 RNAP protein expressed in IBRST7 cells was analyzed by RT-PCR, respectively, the results showed the T7 RNAP gene had been integrated into the chromosome of IBRS-2 cell and expressed stably at high level. To study whether T7 RNAP is of transcriptional activity in the established IBRST7 cell line, a plasmid pIERS-EGFP-ET with a reporter gene (EGFP) under control of the T7 promoter was constructed. IRES element from FMDV (for CAP-independent translation) was cloned into plasmid pET-43.1a-c(+) downstream of the T7 promoter sequence, then EGFP gene was cloned in frame downstream of the AUG codon of the FMDV IRES, resulting in the plasmid. IBRST7 cells were transfected with plasmid pIERS-EGFP-ET using lipfection, EGFP was expressed, the results showed the T7 RNAP in IBRST7 cells has transcriptional activity. IBRST7 cell line was directly transfected with linearized full-length cDNA of swine vesicular disease virus (SVDV) HK/70, infectious SVDV was efficiently recovered from the cDNA. The reverse genetic procedure is simplified to a faster, one step protocol to recover RNA virus and will be useful to understand the mechanisms of molecular pathology of RNA virus and develop effective vaccines.

OBJECTIVEStudy blood vessel injury and gene expression indicating vascular endothelial cell apoptosis induced by mannitol with and without administration of anti-oxidative vitamins.

METHODSHealthy rabbits were randomly divided into four groups. Mannitol was injected into the vein of the rabbit ear in each animal. Pre-treatment prior to mannitol injection was performed with normal saline (group B), vitamin C (group C) and vitamin E (group D). Blood vessel injury was assessed under electron and light microscopy. In a second experiment, cell culture specimen of human umbilical vein endothelial cells were treated with mannitol. Pre-treatment was done with normal saline (sample B), vitamin C (sample C) and vitamin E (sample D). Total RNA was extracted with the original single step procedure, followed by hybridisation and analysis of gene expression.

RESULTSIn the animal experiment, serious blood vessel injury was seen in group A and group B. Group D showed light injury only, and normal tissue without pathological changes was seen in group C. Of all 330 apoptosis-related genes analysed in human cell culture specimen, no significant difference was seen after pre-treatment with normal saline, compared with the gene chip without pre-treatment. On the gene chip pre-treated with vitamin C, 45 apoptosis genes were down-regulated and 34 anti-apoptosis genes were up-regulated. Pre-treatment with vitamin E resulted in the down-regulation of 3 apoptosis genes.

CONCLUSIONVitamin C can protect vascular endothelial cells from mannitol-induced injury.

Animals ; Antioxidants ; pharmacology ; Apoptosis ; Endothelial Cells ; cytology ; pathology ; Gene Expression Regulation ; Humans ; Mannitol ; chemistry ; pharmacology ; Nucleic Acid Hybridization ; Oligonucleotide Array Sequence Analysis ; Oligonucleotide Probes ; chemistry ; Oxidation-Reduction ; Rabbits ; Vitamins ; metabolism

BACKGROUNDDocetaxel (DOC) therapy is well tolerated and shows high response rates in patients with hormone refractory prostate cancer (HRPC). There are many reports on the effect of rapamycin (RPM) on the treatment of carcinogenesis. The goal of this study was to test whether RPM could enhance the susceptibility of both androgen-dependent and -independent prostate carcinoma cells to DOC.

METHODSProstate cancer (PC) cell lines (LNCap, PC3 and AILNCap) were cultured and treated with RPM and DOC alone or in combination. The effects of therapeutic agents on cells were determined by the WST-1 assay. Apoptosis induction was confirmed by flow cytometric analysis. The apopcyto caspase colorimetric assay kit was applied to measure the activities of caspases 3 and 9. The antitumor effects of RPM and DOC against PC cells were also assessed in nude mice using four randomized groups: control, RPM, DOC and combination drug therapy by measuring tumor size. All the animals tolerated both RPM and DOC without significant weight loss.

RESULTSRPM and DOC caused dosage-dependent growth suppression of PC cells. RPM could increase the susceptibility of PC cells to DOC significantly, and combined treatment with RPM and DOC caused synergistic growth suppression in all examined PC cell lines by isobolographic analysis. Both RPM and DOC significantly induced apoptosis in a dosage-dependent manner. RPM (10 nmol/L), DOC (1 nmol/L), and combined treatment induced apoptosis rate were 8%, 17% and 38%, respectively (the control was 2%). RPM could promote the apoptosis induced by DOC in PC cell lines. Both RPM and DOC significantly increased the caspase activity in a dosage-dependent manner. The relative activities of caspase 9 in control, RPM, DOC and RPM + DOC groups were 0.22 +/- 0.02, 0.36 +/- 0.06, 0.47 +/- 0.05 and 0.84 +/- 0.08, respectively. The relative activities of caspase 3 were 0.21 +/- 0.02, 0.24 +/- 0.05, 0.42 +/- 0.06 and 0.81 +/- 0.09, respectively. Either RPM or DOC alone significantly inhibited the growth of PC cells in nude mice compared to the control. The combination of RPM and DOC produced a significant reduction in tumor volume when compared to RPM or DOC alone. After 5-week treatment, the tumor sizes of LNCap in control, RPM, DOC and RPM + DOC groups were (570 +/- 56) mm(3), (412 +/- 41) mm(3), (425 +/- 46) mm(3) and (221 +/- 26) mm(3), respectively.

CONCLUSIONSRPM could significantly increase the susceptibility of both androgen-dependent and -independent PC cells to DOC; the synergy of RPM and DOC was demonstrated. RPM enhanced the DOC-induced upregulation of caspase activity, resulting in an increasing number of cells in sub-G1 phases. The synergy of the combined treatment might be observed in both androgen-dependent and -independent PC cell lines.

Animals ; Antineoplastic Agents ; therapeutic use ; Cell Line, Tumor ; Drug Synergism ; Flow Cytometry ; Humans ; Male ; Mice ; Mice, Nude ; Prostatic Neoplasms ; drug therapy ; Random Allocation ; Sirolimus ; therapeutic use ; Taxoids ; therapeutic use ; Xenograft Model Antitumor Assays

Objective: To analyze the epidemiological characteristics of coronavirus disease 2019 ( COVID-19 ) clusters in Lishui, so as to provide basis for the prevention and control of COVID-19 clusters. Methods: The data of COVID-19 clusters in Lishui from January 23 to March 29, 2020 were collected through China Disease Control and Prevention Information System-Public Health Emergency Information System, and analyzed time, space, scale, source of infection, exposure and transmission route by descriptive epidemiological method. Results: There were 31 cases in 8 clusters ( about 4 cases per cluster ), with no death. The report time was bimodal, peaked first from January 20 to February 10 with 4 clusters imported from domestic and peaked second from March 1 to 29 with 4 clusters imported from overseas. Qingtian County reported 4 clusters, Liandu District, Yunhe County, Qingyuan County and Jingning County each reported 1 cluster. Thirteen cases were restaurant employees, accounting for 41.94%. The cases were mainly occurred in the condition that exposed in the same family ( 6 clusters ), in the same dinner and car ( 1 clusters ), and in the same party ( 1 clusters ). The exposure modes that caused more cases infected were through the same family (9 cases) and through the same dinner and car ( 6 cases ). There were 3 clusters with first-generation cases, 3 clusters with second-generation cases and 2 clusters with third-generation cases. The recurrence rate of the 8 clusters ranged from 1.49% to 7.69%, with a median of 3.47%. Conclusions The COVID-19 clusters in Lishui imported from domestic in the early stage and later from overseas. Most cases were reported from Qingtian County, were engaged in catering business, and exposed by living with families.

OBJECTIVETo evaluate the significance of determining ascitic bacterial 16S rRNA by quantitative PCR combined with microarray (PCR-microarray) in the diagnosis of spontaneous bacterial peritonitis (SBP).

METHODSAscitic bacterial 16SrRNA was determined by real time fluorescent quantitative PCR-microarray in 76 cases of suspected SBP and 6 cases of non-infectious ascites with chronic liver diseases. The results were compared with ascitic bacterial culture simultaneously.

RESULTSOf 76 ascitic samples, 17 were detected bacteria positive by PCR-microarray, including 8 Grams positive(G+) and 9 Grams negative(G-), which was higher than that by bacterial culture which had only 6 ascitic samples detected positive (all G-); the positive rates were 22.4% vs 7.9%, respectively (P < 0.01). The bacterial strains detected by both methods in 6 cases had a consistency with each other. No bacteria were detected in another 6 cases of non-infectious ascites with chronic liver diseases.

CONCLUSIONSDetermination of ascitic bacteria 16S rRNA by PCR-microarray has a higher specificity and sensitivity in the diagnosis of SBP as compared with the bacteria culture. Application of this novel method can not only accelerate SBP diagnosis but also stratify the different pathogens.

Adult ; Aged ; Ascitic Fluid ; microbiology ; Bacterial Infections ; diagnosis ; microbiology ; Female ; Humans ; Liver Cirrhosis ; diagnosis ; microbiology ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Peritonitis ; diagnosis ; microbiology ; Polymerase Chain Reaction ; methods ; RNA, Bacterial ; isolation & purification ; RNA, Ribosomal, 16S ; isolation & purification

OBJECTIVETo analyse the relationships among HBV genotypes, HBV DNA levels and histopathological features of the livers of familial grouped hepatitis B patients in the Tianjin area.

METHODSOne hundred familial grouped hepatitis B (CHB) patients were enrolled in this study. Thirty-five of the 100 patients were chronic asymptomatic HBsAg carriers (ASC) and 65/100 were mild CHB patients. Their HBV genotypes and HBV-DNA levels were detected and liver biopsies were performed for analyzing the pathological features.

RESULTSSeven patients were of HBV genotype B (7%), and most of them had a HBV DNA level in the middle 10(3-5) copies/ml (57.14%). The histopathological features of the livers were of different degrees of injury. Eleven patients HBV was of genotype BC (11%); their HBV DNA levels were from 10(3-5) copies/ml (45.45%) to 10(6-7)copies/ml (36.36%). Their liver pathology showed slight or severe injuries (< or = G2 90.91%, < or = S(2) 81.82%). Eighty-two patients HBV was of genotype C (82%), and among the 82, 29 were ASC and 53 were CHB. Among the ASC, most of them had a high HBV DNA level (72.41%), and all of them had different degrees of liver injury. Among the CHB, their HBV DNA levels were 10(6-7) copies/ml (39.62%) and more than or equal 10(8) copies/ml (49.06%). The liver histopathological features were > or = G(2) in 38 patients (71.70%), and > or = S(2) in 25 patients (47.17%).

CONCLUSIONS(1) In the majority, HBV of the family gathered hepatitis B patients living in Tianjin is of genotype C and they have a high HBV-DNA level and severe liver pathological injuries. These features of the family gathered hepatitis B patients are the main factors causing the unfavorable prognosis of the patients. (2) There is inflammation of different degrees in the livers and high HBV DNA levels in all the family gathered ASC patients. Antiviral therapy should be planned according to the pathological features in patients livers. (3) Liver biopsies should be performed routinely before their antiviral therapy.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; DNA, Viral ; Female ; Genotype ; Hepatitis B virus ; genetics ; physiology ; Hepatitis B, Chronic ; genetics ; pathology ; virology ; Humans ; Liver ; pathology ; virology ; Male ; Middle Aged ; Pedigree ; Viral Load ; Young Adult