Objective To investigate the effect of berberine on growth and apoptosis of human cervical cancer cell line HeLa and its possible mechanism.Methods The effect of berberine on growth of HeLa cells was studied by MTT assay.Apoptosis of HeLa cells exposed to berberine was observed by flow cytometry and DNA gel electrophoresis.The expression of Bax and Bcl-2 was studied by Western blotting analysis.Results Berberine markedly inhibited the proliferation of HeLa cells in a time-and dose-dependent manner.After incubation of HeLa cells with 20 and 40 mg/L berberine for 48 h,DNA Ladder can be observed.A typical "sub-G1 peak" was checked by flow cytometry.There was a very low rate of natural apoptosis(1.9?0.6)%,while in 5 mg/L berberine group,the apoptosis rate was(2.3?0.8)%.After exposing HeLa cells for 48 h to 20 and 40 mg/L berberine,the apoptosis rate reached(16.7?2.8)%(P

Objective To investigate the significance of non-bronchial systemic collaterals (NBSCs) in supplying bronchial hemoptysic lesions,and to study the morphological features of bronchial artery (BA) when NBSCs become the predominant supplying vessels. Methods Multi-slice helical CT angiographic findings in 124 patients with bronchial hemoptysis were retrospectively analyzed. 3D reconstruction of thoracic systemic arteries,including BAs and NBSCs,was performed at the console work station with the help of real-time thin-slice enhanced helical CT scanning. The number of NBSCs and BAs was calculated,and the internal diameter of the arteries and the thickness of pleura in the vicinity of the pulmonary lesion were measured. According to the presence or absence of NBSCs,the patients were divided into NBSCs group and non-NBSCs group. The relevant data was statistically analyzed. Results NBSCs group included 36 cases,the mean internal diameter of BA was (1.850 ? 0.631)mm and the pleura adjacent to the pulmonary lesion was obviously thickened in 22 cases (61%) with a thickness of 2.7-16.0 mm [mean(7.71 ? 4.12) mm]. In the non-NBSCs group (n = 88),the mean internal diameter of BA was (2.200 ? 0.528) mm and the pleura adjacent to the pulmonary lesion was obviously thickened in 7 cases (8%) with a thickness of 1.1-2.4 mm [mean(1.7 ?0.53) mm]. The differences in both internal diameter of BA and the thickness of lesion's adjacent pleura between two groups were statistically significant (P

Objective To evaluate the utility of CD4+ TCR tetramers‐based flow cytometric analysis and cell climbing slice assay in detecting antigen‐specific CD14+ monocytes in the blood of tuberculosis (TB) patients .Methods CD4+ TCR tetramers were used to detect tetramer‐positive CD14+ monocytes in the peripheral blood (PBL ) samples of inpatients with advanced pulmonary TB (PTB) by flow cytometric analysis .The PBL samples obtained from non‐TB patients and umbilical cords were used as controls .These tetramers were also used to examine tetramer‐bound CD14+ monocytes and Mycobacterium tuberculosis (MTB) antigen‐specific and tetramer‐bound cells by cell climbing slice in situ staining .Results The median percentage of tetramer‐bound CD14+ monocytes in PBL samples from PTB patients ,non‐TB patients and umbilical cords were 1 .32% , 0 .50% and 0 .26% respectively by using CD4+ Vα21‐J39/Vβ29‐D1‐J2 tetramer , while the medians were 1 .05% , 0 .49% and 0 .19% respectively by using CD4+ Vα21‐J39/Vβ29‐D2‐J2 tetramer . The percentage of tetramer‐bound CD14+ monocytes in PTB patients group was significantly higher than the other two control groups .In cell climbing slice in situ staining ,tetramer‐bound CD14+ monocytes ,and MTB antigen‐specific and tetramer‐bound cells were positive in PTB tissue compared with negative in control tissues . Conclusions CD4+ TCR tetramers‐based flow cytometric analysis and cell climbing slice assay could be used to sensitively detect M TB antigen‐specific CD14+ monocytes in the blood of TB patients ,and more accurately evaluate the changing profile and clinical significance of these cells in TB patients .

Objective To investigate the renal angiographic manifestations of severe hemorrhage following minimally invasive pereutaneous nephrostolithotomy (MPCNL), and to evaluate the technique of super-selective renal arterial embolization in treating the condition. Methods Forty-eight cases of severe hemorrhage following MPCNL treated with super selective renal arterial embolization in our department were retrospectively reviewed. The angiographic findings, results and complications of embolization procedures were analyzed. Results Two cases were of acute hemorrhage immediately after MPCNL, and the other 46 cases were of delayed hemorrhage 2 to 7 days after MPCNL. Of these 48 cases, 25 (52.1%) showed simple pseudo-aneurysms, 6 (12.5%) pseudo-aneurysms accompanied with arterial-venous shunts, 1 (2.1%) pseudo-aneurysm with extravasated contrast medium, 11 (22.9%) arterial-venous fistulas, 2 (4.2%) extravasated contrast medium from arterial branches, 1 (2.1%) renal capsular branches varix, 2 (4.2%) no lesion detected. Successful super-selective embolization was achieved in all 46 positive cases, and renal hemorrhage was stanched consequently. Polyvinyl alcohol foam embolization particles (PVA), gelfoam and coils were used in the procedures (PVA in 18 procedures, PVA +coil in 5, gelfoam in 10, geffoam + coil in 11, PVA + gelfoam + coil in 2). Post-embolization syndrome of various degrees were seen in all treated patients. A slight rise in blood creatinine levels was observed in 12 cases. Conclusion Super selective renal arterial angiography and embolization is the treatment of choice in patients who suffered severe hemorrhage due to MPCNL.

Colorectal cancer (CRC), one of the most common malignant tumors, is caused both by environmental and genetic factors. Genetic factor plays an important role in the pathogenesis of CRC. Genome-wide association study (GWAS) is a new tool of genetic research. A series of susceptibility genes and loci of the complex diseases has been identified with GWAS strategy. In this article, the research progress on GWAS of CRC is reviewed, and the advantages and limitations of GWAS study as well as the prospective of its application are discussed.
Colorectal Neoplasms ; genetics ; Genetic Predisposition to Disease ; Genome-Wide Association Study ; Humans ; Polymorphism, Single Nucleotide

Objective:To establish a method of multi-PCR to amplify the complete DNA sequence (CDS) of TCR ? and ? chain of the antigen-specific T lymphocytes in local pathologic specimen of active pulmonary tuberculosis patients, and to analyze ?/? T cell receptor gene rearrangement and CDR3 repertoire.Methods:The lymphocytes in bronchoalveolar lavage (BAL) of active pulmonary tuberculosis patients were separated. Following total RNA extraction, cDNA synthesis, Multi-PCR, recombinant clones construction, and sequencing, the CDS of TCR ? and ? chains from these lymphocytes were analyzed by using software of DNAstar and internet TCR resources.Results:24 of ? chain CDS and 13 of ? chain CDS from 3 samples of BAL were obtained. As for TCR ? chain, AV1S2 (54%), AV12S3 (41%), and AV12S2(5%) appeared frequently. BV2(38%), BV29S1(46%), BV14(3%), and BV4S2(3%) in TCR ? chain appeared more often. There were CDR3 diversities between samples and even in the same sample by amino acid sequence analysis, but there were a few identical or similar amino acid sequences. There was the same amino acid sequence of SVGTGTLHQETQY in CDR3 region of ? chain of BAL sample No.1 and No.2; The sequence of AVRDWAGNMLT appeared in two ? chains of BAL sample No.2 and No.3; Moreover, the sequence of AV…DNN…RLM appeared in ? chains of BAL sample No.2 and No.3.Conclusion:A method of Multi-PCR is used to amplify TCR ? and ? chain CDS of tuberculosis patients. There are characteristic T cell clones to proliferate,with TCR ? and ? chain repertiore skewing in local infective focus. The sequences of CDR3 in different TCR clones are mostly different but there are a few identical or similar sequences in the same patient or even between different patients. The identical amino acid sequences of CDR3 are possibly specific for recognizing MTB polypeptide.

Objective To construct and apply a cell line screening Mycobacterium tuberculosis (Mtb)-specific tetramers of CD4+α/β T cell receptor(TCR). Methods The β chains of HLA class Ⅱ (DR) were amplified from tuberculosis patients by PCR. The pMT-HLA-DRB expression vectors that carries the HLA-DR 13 chain and pMT-HLA-DRA-P expression vectors which carries the genes of HLA-DR α chain loaded with Mtb antigen were transfected into S2 cells with the method of calcium phosphate transfection. The expressed Mtb peptide/HLA-DR complexes were primarily identified by the method of cell immunohistochemistry. The cell lines expressing Mtb peptide/HLA-DR complexes were used to screen tetramers of CD4+ TCR by flow cytometry. Results S2 cell lines expressing Mtb peptide/HLA-DR complexes on the cell surface were obtained, two kinds of Mtb specific tetramers of CD4+α/β TCR were screened. Conclusion S2 cell lines expressing Mtb peptide/HLA-DR complexes on the cell surface provide the solid basis of the further research on the TCR tetramers and are helpful for exploring new diagnostic study methods about tuberculosis and developing new vaccines.

ObjectiveTo investigate the specificity of CD4+ Vα9-J27/Vβ29-D1-J2 tetramer in detecting Mycobacterium tuberculosis(MTB) infections.MethodsThe above TCR tetramer by using biotinylated monomers expressed and purified from constructed stable Drosophila Schneider 2 cell( S2 cell) lines was prepared.The PE-labled TCR tetramer was used to costain with S2 cell lines expressing MTB prptide/HLA-DR complexes on the cell membrane,and also was used to detect tetramer-bound CD14+ monocytes and macrophages in the peripheral blood mononuclear cells (PBMC) of pulmonary tuberculosis (PTB) patients and three control groups by flow cytometric analysis.And the FITC-labled tetramer was used to examine tetramer-bound CD14+ monocytes and macrophages,and MTB antigen-specific and tetramer-bound cells by in situ staining.ResultsThe TCR tetramer was well binding with S2 cell lines expressing C14/HLA-DR *1504 on the cell membrane.By flow cytometric analysis,the percentage of tetramer-bound CD14+ monocytes and macrophages in PTB patients group was higher than the other three control groups( P＜0.001 ).By in situ staining,tetramer-bound CD14+ monocytes and macrophages,and MTB antigen-specific and tetramer-bound cells were positive in PTB tissue and negative in control pneumonia tissue.ConclusionThe spcificity of TCR tetramer in monitoring MTB infections by flow cytometric analysis and in situ staining could be seen,which laid a laboratory foundation in the diagnosis and immune mechanism research of TB by using TCR tetramers.

Cardiovascular diseases (CVDs), including coronary artery disease, stroke, heart failure, and hypertension, are the global leading causes of death, accounting for more than 30% of deaths worldwide. Although the risk factors of CVDs have been well understood and various treatment and preventive measures have been established, the mortality rate and the financial burden of CVDs are expected to grow exponentially over time due to the changes in lifestyles and increasing life expectancies of the present generation. Recent advancements in metagenomics and metabolomics analysis have identified gut microbiome and its associated metabolites as potential risk factors for CVDs, suggesting the possibility of developing more effective novel therapeutic strategies against CVD. In addition, increasing evidence has demonstrated the alterations in the ratio of Firmicutes to Bacteroidetes and the imbalance of microbial-dependent metabolites, including short-chain fatty acids and trimethylamine N-oxide, play a crucial role in the pathogenesis of CVD. However, the exact mechanism of action remains undefined to this day. In this review, we focus on the compositional changes in the gut microbiome and its related metabolites in various CVDs. Moreover, the potential treatment and preventive strategies targeting the gut microbiome and its metabolites are discussed.

Objective Interleukin-27 (IL-27) has been reported to reduce the levels of interleukin17(IL-17) and alleviate the severity of experimental autoimmune myocarditis.IL-17,an important tissueprotective cytokine in viral myocarditis (VMC),has been reported to increase synovial expression of IL-27 in rheumatoid arthritis.However,the influence of IL-17 on IL-27 expression in murine model of VMC remains unknown.Methods Wild-type (WT) and IL-17A-deficient (IL-17A-/-) mice on the BALB/c background were intraperitoneally (i.p) injected with coxsackievirus B3 (CVB3) for establishing VMC models.Cardiac tissue was obtained on day 7 after CVB3 injection.Myocardial histopathologic changes were observed by hematoxylin-eosin (HE) stained myocardial sections.Expression of IL-27 in heart and serum was measured by real-time reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA),respectively.Furthermore,splenic lymphocytes and peritoneal macrophages were purified 1 week after injection from WT mice.Isolated lymphocytes were cultured in the presence of different concentrations (0 and 25 ng/ml) of recombinant IL-17 (rIL-17) for 24 h.Macrophages were cultured with different concentrations of rIL-17 (0 and 10 ng/ml) for 48 h.IL-27 mRNA expression of cultured cells was assayed by RT-PCR,and their protein level in the culture supernatant was measured by ELISA.Results Compared with WT mice,significantly less cardiac inflammation was evidenced in the heart of IL-17A-/-mice (0.9 ± 0.3 vs.1.9 ± 0.5),relative cardiac IL-27 p28 mRNA expressions (1.11 ± 0.24 vs.3.1 ± 0.8) and serum IL-27 protein [(72 ± 18) pg/ml vs.(95 ± 25) pg/ml] were also significantly lower in IL-17A-/-mice (all P ＜ 0.05).In the culture lymphocytes,the relative mRNA (1.02 ± 0.13 vs.1.32 ± 0.21) and protein [(49 ± 9) pg/ml vs.(52 ± 11) pg/ml] expressions of IL-27 p28 and were similar post treatment with 0 and 25 ng/ml rIL-17 (all P ＞0.05).Compared with 0 ng/ml rIL-17 culture with macrophages,higher relative mRNA (8.5 ±3.1 vs.2.2 ±0.7) and protein [(368 ±95) pg/ml vs.(150 ± 38) pg/ml] expressions of IL-27 p28 were detected in 10 ng/ml rIL-17 group (all P ＜0.05).Conclusion Our data indicates that cytokine IL-17 may contribute to the secretion of IL-27 in VMC mice.Furthermore,macrophages but not lymphocytes may be the important IL-27-producing immune cells and major target cells for IL-17.Thus,IL-27 and IL-17 might be actively involved in the pathogenesis of VMC.