The Changes of Hepatitis C Virus Gene Subtypes and RNA Loads in the Development of Chronic Hepatitis C Patients in Yunnan / 昆明医科大学学报

Objective To investigate the changes of hepatitis C virus (HCV) gene subtypes and RNA loads in chronic hepatitis C, HCV－related liver cirrhosis and HCV－related hepatocellular carcinoma patients in Yunnan. Methods 241 patients were enrolled at the First Affiliated Hospital of Kunming Medical University (Yunnan, China) from January 2016 to April 2017. Among them, 169 patients were with chronic hepatitis C, 56 patients with HCV－related liver cirrhosis and 16 patients with HCV－related hepatocellular carcinoma. HCV gene subtype and RNA loads were measured using the Polymerase Chain Reaction－fluorescence probe method. Results In the chronic hepatitis C group,there were 47 subtype 3b cases (27.81%) . 17 cases of HCV－related liver cirrhosis were subtype 1b (30.36%);5 patients with HCV－related hepatocellular carcinoma were subtype 3b (31.25%) . There was no statistical difference distribution of the genotype among the three groups (P＞0.05) .The HCV RNA loads of the chronic hepatitis C group, HCV － related liver cirrhosis group and HCV － related hepatocellular carcinoma group were (332±114) copies/mL, (189±73) copies/mL and (152±56) copies/mL respectively. The difference among three groups were significant (P＜0.01).The chronic hepatitis C group was significantly higher than the other two groups, and the differences were statistically significant (P＜0.01) . But no significant difference of HCV RNA loads was found between HCV － related liver cirrhosis and HCV － related hepatocellular carcinoma group (t=0. 65,P＜0.05) . Conclusion In Yunnan, 3b was main genotype in chronic hepatitis C patients and 1b was main genotype in HCV－related liver cirrhosis, 3b was main genotype in HCV－related hepatocellular carcinoma. HCV RNA loads tend to decrease in the progress that chronic hepatitis C develops into HCV－related liver cirrhosis and HCV－related hepatocellular carcinoma.

Establishment and characterization of lung adenocarcinoma cell line XLA-07 / 中华病理学杂志

<p><b>OBJECTIVE</b>To establish and characterize a lung adenocarcinoma cell line from a female patient in Xuanwei, Yunnan province.</p><p><b>METHODS</b>Surgical specimen of the lung adenocarcinoma was obtained and cultured immediately in RPMI 1640 medium with 10% fetal bovine serum and 10(5) U/L penicillin and 100 mg/L streptomycin. When stable proliferation of the cells was achieved after over 40 passages in culture, the biological features of the cell line were investigated by cell morphology, karyotyping, protein marker expression [cytokeratins (CKs), epithelial membrane antigen (EMA) and CD proteins], growth kinetics, cell cycle phase distribution, mitotic index, colony formation in soft agar, cell invasion and tumorigenicity in Balb/c nude mice.</p><p><b>RESULTS</b>The established cell line was stably cultured for over 80 passages during a one-year period as an anchorage-dependent monolayer of short spindle, polygonal to epithelioid cells under phase contrast microscope. Microglandular cavities and disordered microfilaments were observed under transmission electron microscope. The growth curve presented in an "S" shape with the cell population doubled every 46.7 hours. The mitotic index was 1.5% and the colony formation rate was 8.3%. The cell cycle distribution included 76.9% in G(0)/G(1), 15.1% in S and 8.0% in G(2)/M. The cell line displayed a hypotriploid karyotype with a mode of 66 chromosomes and a median of 64 chromosomes. The cells expressed CK7, CK8, CK (Pan) and EMA by immunohistochemistry. A high level of cell surface expression of CD13 and CD59 was evident by flow cytometry. The cells were able to penetrate Matrigel in vitro but failed to form a stable xenograft in nude mice.</p><p><b>CONCLUSION</b>A new human lung adenocarcinoma cell line, designated as XLA-07, is successfully established from a Xuanwei lung cancer patient.</p>

Role of protein kinase C-delta in hyperthermia-induced apoptosis in tongue squamous cell carcinoma Tca8113 cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the role of protein kinase C-delta (PKC-delta) in hyperthermia-induced apoptosis in human tongue squamous cell carcinoma Tca8113 cells.</p><p><b>METHODS</b>Tca8113 cells were treated at 43 degrees C in a heating water bath for 0, 40, 80, 120 min after pretreatment with Rottlerin, a specific inhibitor of PKC-delta, and equal volume dimethyl sulfoxide (DMSO) for 30 min, respectively. The cells were stained by propidium iodide (PI) and Rhodamine 123 to analysis apoptotic rate and the changes of mitochondrial transmembrane potential by flow cytometry (FCM). The total proteins were extracted for Western blotting analysis of activation and proteolysis of PKC-delta, and for colorimetric assay of relative activity of Caspase-3.</p><p><b>RESULTS</b>Hyperthermia could induce proteolysis and activation of PKC-delta, and this was attenuated by Rottlerin. Apoptotic rate, decreasing of mitochondrial transmembrane potential and activity of Caspase-3 which being induced by hyperthermia in Tca8113 cells were inhibited by PKC-delta specific inhibitor Rottlerin. There were significantly statistical differences in apoptosis rates, mitochondrial transmembrane potential and activity of Caspase-3 between Rottlerin- and non-Rottlerin-pretreated cells after hyperthermia for 40, 80, 120 min (P < 0.01).</p><p><b>CONCLUSION</b>Activated PKC-delta may facilitate hyperthermia-induced apoptosis in Tca8113 cells, and may be one of the mechanisms of apoptosis induced by hyperthermia.</p>

Study on the distribution of vitamin D receptor gene start codon polymorphism in the Achangs and Hans / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect the difference between the Chinese Achang and Han ethnic groups in Yunnan province in the distribution of vitamin D receptor (VDR) gene start codon polymorphism.</p><p><b>METHODS</b>Polymerase chain reaction-restriction fragment length polymorphism, gene sequencing and genetic analysis methods were used. A restriction fragment length polymorphism in the start codon of VDR (Fok I) gene was tested in the Achangs (n=68) and the Hans (n=92).</p><p><b>RESULTS</b>The frequencies of FF, Ff and ff genotypes were found to be 18%, 35% and 47% in the Achangs, and 22%, 52% and 26% in the Hans, respectively. A significant difference was seen in the frequency distribution of VDR genotype between the Achangs and the Hans(Chi2=7.716, P=0.021).</p><p><b>CONCLUSION</b>The Achang and Han ethnic groups differ in the frequency distribution of VDR gene start codon polymorphism.</p>