Identification of HBx-related integration sites in HBsAg-positive hepatocellular carcinoma biopsy / 中华肝脏病杂志

To identify the integration sites in the host genome for the hepatitis B virus (HBV)-encoded X protein (HBx) in hepatocellular carcinoma (HCC) biopsies that are positive for hepatitis B surface antigen (HBsAg). HCC biopsies were obtained from six patients that were HBV carriers, as demonstrated by the presence of HBsAg in their serum and sero-negativity for antibody to HBsAg. DNA was extracted from the tissue, fractionated, and circularized. Primers were designed according to the HBx sequence and used to amplify the circularized DNA templates by inverse polymerase chain reaction (IPCR). The amplified DNA fragments were checked by electrophoresis, cloned into the PMD18-T expression vector, and sequenced. Sequence alignment was performed by the Blast algorithms. Seven electrophoresis bands yielded 22 sequencing results, which represented a total of three HBx integration sites in the host genome: 19q12, 2q32.2, 22q12. The 19q12 integration site encompasses the CCNE1 gene, which encodes a G1/S-specific cyclin-E1. HBx-related integration sites exist in HBsAg-positive HCC biopsies. The CCNE1 gene may play a role in the development of HBx-related HCC.

Comparision of laparoscopic and open left lobectomy: a prospective controlled study / 中华消化外科杂志

ObjectiveTo compare the efficacy of laparoscopic and open left lobectomy.MethodsThe clinical data of 92 patients who received left lobectomy at the First Affiliated Hospital of Harbin Medical University from May 2010 to June 2011 were retrospectively analyzed.Of the 92 patients,42 received laparoscopic left lobectomy (laparoscopic group ) and 50 received open left lobectomy (open group ). The advantages and disadvantages between laparoscopic and open left lobectomy were compared. All data were analyzed using the t test,chi-square test or by calculating the Fisher exact probability.ResultsTwenty-nine patients received left lateral lobectomy and 13 patients received left hemihepatectomy in the laparoscopic group. One patient was converted to the open group becaused of the injury of the middle hepatic vein.Thirty-three patients nnderwent left lateral lobectomy and 17 underwent left hemihepatectomy in the open group.The tumor-free resection margin of the laparoscopic group was ( 1.6 ± 0.6 ) cm,which was significantly longer than ( 1.2 ± 0.4 ) cm of the open group (t=3.81,P＜0.05).The volume of operative blood loss of the laparoscopic group was (158 ±89)ml,which was significantly smaller than (292 ± 172)ml of the open group (t =4.56,P ＜ 0.05 ).The time of postoperative pain control,time to bowel function recovery and duration of hospital stay were ( 1.2 ± 0.3 )days,(23 ± 4)hours,( 7.5 ± 2.8 ) days in the laparoscopic group,which were significantly shorter than ( 2.0 ± 1.1 ) days,(4.9 ± 7 ) hours,( 11.3 ± 4.2 ) days in the open group,respectively ( t =4.57,21.31,5.00,P ＜ 0.05 ).The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) at postoperative day 1 were increased,while the increase of AST and ALT in the open group were greater than that in the laparoscopic group (t =6.73,5.03,P ＜0.05).The postoperative prothrombin time in the open group was significantly longer than that before operation (t =2.32,P ＜ 0.05 ).The incidence of postoperative complications and total hospital costs were 7％ (3/41) and (2.5 ±0.7) ×104 yuan in the laparoscopic group,which were lower than 8％ (4/50) and (2.6 ±0.6) × 104 yuan in the open group,but no significant difference was observed (t =0.74,P ＞0.05).One patient in the open group died of multi-organ dysfunction syndrome caused by acute hepatic failure.ConclusionLaparoscopic left lobectomy is safe and effective,and it has the advantages of small trauma,quick recovery of patients and significant overall efficacy when compared with open left lobectomy.

Effect of Saikosaponins-d on reversing malignant phenotype of HepG2 cells in vitro / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To observe the effect of SSd on reversing the malignant phenotype of HepG2 cells and to investigate its mechanism in order to prove that SSd is a new choice to prevent and treat HCC.</p><p><b>METHODS</b>HepG2 cells were cultured and treated by different concentration (0 mg/L, 2.5 mg/L, 5.0 mg/L, 10.0 mg/L and 20.0 mg/L) of SSd for 24 h, and treated by 10 mg/L of SSd for 0 h, 6 h, 12 h, 24 h, 48 h and 72h respectively. The cell inhibition rates were measured by MTT assay. Then cells were treated by 10 mg/L SSd for 48 hr in experimental group and treated by no SSd as a control, their morphological changes were observed by contrast phase microscope. The concentrations of ALB and AFP in clear supernatant liquid of cells were detected by radio-immunity and chemiluminescence. The cell migration rates were observed by transwell method, the relative expression levels of p27 mRNA were measured by RT-PCR.</p><p><b>RESULTS</b>The inhibitive effect of 10 mg/L SSd was the most significant among different concentrations ( F = 265.06, P less than 0.01). The shape of HepG2 from experimental group turned into small and round, and their volume ratios of nucleus to plasma decreased. ALB in supernatant liquid of HepG2 was higher ( t = 7.83, P less than 0.05, and its AFP was lower ( t = -10.72, P less than 0.01) as compared to control group. Cells migrated were fewer and p27 mRNA expression of HepG2 was higher in experimental group than that in control group (t = 22.00, P less than 0.05).</p><p><b>CONCLUSION</b>SSd could reverse the malignant phenotype of HepG2 cells. It was suggested that the up-regulation of p27 mRNA expression play an important role in the differentiation of HepG2 cells treated by SSd.</p>

Integration sites in HCC biopsy / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To compare the performance of Inverse-PCR, Alu-PCR and Cassette-ligation-mediated PCR (CLM-PCR) in HBV DNA integration sites identification.</p><p><b>METHODS</b>One HCC biopsy was obtained from surgically resected sample. The patient was positive for serum hepatitis B surface antigen (HBsAg). The genomic DNA was purified by the standard phenol/chloroform extraction and ethanol precipitation method. Seperated set of primers were designed to amplify the HBV DNA integration region by means of 3 different PCR methods respectively. The PCR products were analyzed by electrophoresis, then cloned to PMD18-T vector for DNA sequencing. The sequence alignment was performed under Blast software.</p><p><b>RESULTS</b>7 bands and 22 sequencing results was obtained from IPCR and 3 integration sites was identified. Alu-PCR provided 12 bands and 32 sequencing results, and CLM-PCR showed 12 bands and 4 sequencing results. No integration site was identified from the latter two.</p><p><b>CONCLUSION</b>IPCR compared with another two methods showed a reliable capacity in HBV DNA integration site identification.</p>

The clinical and genetic studies in a family of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the clinical features and genetic diagnostic method of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL).</p><p><b>METHODS</b>A systematic study on the clinical manifestations, neuroimaging characteristics, therapeutic measures and molecular genetics was performed. An investigation on the onset and hereditary pattern of the family was also done.</p><p><b>RESULTS</b>The main clinical features including poor memory and history of stroke were found. And no risk factors of hypertension and arteriosclerosis were found. A positive family history was confirmed. Neuroimaging examination showed multiinfarct lesions and leukoencephalopathy. All these features are in conformity with those of CADASIL. A mutation in the third and fourth exon of the NOTCH3 gene was identified in the 10 cases of 4 generations. The clinical or subclinical onset in the 10 cases was consistent with classical autosomal dominant inheritance.</p><p><b>CONCLUSION</b>The clinical and molecular genetic features of the family accord with CADASIL.</p>

The expression of caspase-10 in differentiated thyroid carcinoma and association with its development and metastasis / 中国地方病学杂志

Objective To investigate the expression of caspase-10 in differentiated thyroid carcinoma and association with its development and metastasis. Methods Thyroid samples from 37 patients in a period from January 2006 to December 2007, with differentiated thyroid carcinoma were retrospectively analyzed for caspase-10 by immunohistocbemistry(streptavidin-perosidase, S-P), compared to control group of 46 cases with nodtdar goiter. The relationship between the expression of caspase-10 and the clinical pathologic characteristics of thyroid carcinoma were also explored simultaneously. Results caspase-10 were observed as brown or yellow particles located in the cytoplasm or cell membrane of nodular goiter but there were no significant evidence for its positive expression in thyroid carcinoma, caspase-10 expression was markedly down-regulated in differentiated thyroid carcinoma(29.73%,11/37) compared with benign nodules(71.74%,33/46, χ2=14.528, P<0.01). The positive expression in 18 cases with lymph node metastasis(11.11%,2/18) was significantly lower than those in 19 patients without lymph node metastasis(47.37%,9/19; χ2=4.210, P<0.01). There was no significant correlation(P> 0.05) between the expression of caspase-10 and the clinical pathologic characteristics including male, age, TNM stage and pathologic type. Conclusion Down-regulation of caspase-10 may play a critical role in carcinogenesis and development of differentiated thyroid carcinoma.

Studies on chemical constituents from rhizome of Anemone flaccida / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents from Anemone flaccida.</p><p><b>METHOD</b>Chemical constituents were isolated by repeated column chromatography (silica gel, Toyopearl HW-40C and preparative HPLC). The structures were elucidated on the basis of spectral data analysis.</p><p><b>RESULT</b>Twelve triterpenes were isolated and their structures were identified as follow: oleanolic acid (1), oleanolic acid 3-O-beta-D-glccopyranosyl-(1-->2)-beta-D-xylopyranoside (2), eleutheroside K (3), oleanolic acid 3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-xylopyranoside (4), oleanolic acid 3-O-beta-D-glccopyranosyl-(1-->2)-alpha-L-arabinofurnoside (5), oleanolic acid 3-O-beta-D-glccuronopyranose (6), oleanolic acid 3-O-beta-D-glccuronopyranose methyl ester (7), oleanolic acid 28-O-alpha-L-rhamnopyranosyl(1-->4)-beta-D-glccopyranosyl (1-->6)-beta-D-glccopyranosyl (8), oleanolic acid 3-O-beta-D-glccuronopyranose 28-O-alpha-L-rhamnopyranosyl (1-->4)-beta-D-glccopyranosyl (1-->6)-beta-D-glccopyranoside (9), oleanolic acid 3-O-beta-D-glccopyranosyl methyl ester 28-O-alpha-L-rhamnopyranosyl (1-->4)-beta-D-glccopyranosyl (1-->6)-beta-D-glccopyranoside (10), oleanolic acid 3-O-beta-D-glccopyranosyl-(1-->2)-beta-D-xylopyranosyl-28-O-alpha-L-rhamnopyranosyl (1-->4)-beta-D-glccopyranosyl (1-->6)-beta-D-glccopyranoside (11), oleanolic acid 3-O-alpha-L-rh-amnopyranosyl-(1-->2)-alpha-L-arabinopyrnosyl-28-O-alpha-L-rhamnopyranosyl (1-->4)-beta-D-glccopyranosyl (1-->6)-beta-D-glccopyranoside (12).</p><p><b>CONCLUSION</b>compounds 5-8, 10, 12 were isolated from this plant for the first time. Compounds 2, 5 and 11 showed positive anti-tumor activities.</p>

Study on prevention and treatment of pancreatic fistula due to pancreatic injury / 中国现代普通外科进展

Objective:To investigate the principle and measures of prevention and treatment on pancreatic fistula due to pancreatic injury.Method:The clinical data of 131 pancreatic injury patients were analyzed retrospectively.Operation and combined therapy during perioperative were performed.Results:Among 35 cases(26.7%,35/131) with pancreatic fistula,3 cases shaped internal fistula showed by visualization and cured spontaneously after operations,9 cases with pancreatic pseudocyst after operations were treated by Roux-en-Y pancreatic cystojejunostomy,2 cases(5.7%,2/35) died of MODS and abdominal infections.The rest were recovered.The follow-up from 3 months to 5 years showed that there were no cases died of complications related to pancreatic fistula.Conclusions:Operation combined with multiple therapies was effective in the prevention and treatment of pancreatic fistula.