Effect of Yingyang Gongji Wan Contained Serum on Inhibiting HepG2 Cell Proliferation and Invasion, and Inducing Apoptosis / 中国实验方剂学杂志

Objective:To investigate the effect of Yingyang Gongji Wan (YYGJ) on hepatoma cell line HepG2, and provide evidence for clinical application. Method:YYGJ-contained rats serum was prepared. Then the inhibiory rate of cells was detected by methye thiazolye telrazlium (MTS) method in both YYGJ group and blank group. Apoptosis of HepG2 was detected by TdT-mediated dUT nick-end labeling (TUNEL) method in blank group,YYGJ group, and 5-fluorouracil (5-FU) group. The mRNA expression and protein expression levels of E-cadherin, Vimentin, metalloproteinase-2 (MMP-2) and Smad4 were detected by Real-time quantitative PCR (Real-time PCR) and Western blot respectively. The invasion ability of HepG2 cells was detected by cell migration assay (transwell). Result:YYGJ-contained serum inhibited the proliferation of HepG2 cells in a time and concentration-dependent manner. As compared with blank group, the inhibitory rate was increased in 5%, 10%, and 20% YYGJ-contained serum groups on the third day (PPPPConclusion:YYGJ-contained serum can inhibit the proliferation of HepG2 cells, induce apoptosis, regulate epithelial-mesenchymal transition (EMT)-related E-cadherin, Vimentin, MMP-2 and Smad4 genes and proteins, and decrease tumor invasion ability. The effect was similar to that of 5-fluorouracil. As a unique prescription, YYGJ can be used as a representative for the treatment of coldness and dampness syndrome of primary liver cancer and its anti-cancer mechanism should be further studied.

Promoter methylation status of hPer3 gene in AML patients and the in vitro effect of decitabine on the status / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the clinical significance of promoter methylation status of hPer3 gene in acute myeloid leukemia (AML) patients and the in vitro effect of decitabine (DCA) on AML cell lines HL-60 and U937.</p><p><b>METHODS</b>The promoter methylation status of hPer3 gene and mRNA expression levels in bone marrow of 206 AML and 40 iron deficiency anemia (IDA) patients (as control) were detected by methylation specific PCR (MS-PCR) and real-time PCR (RT-PCR). The HL-60 and U937 cell lines were treated with different concentrations of DCA for 48 and 72 h. The inhibition rates of cell proliferation were detected by methyl thiazolyl tetrazolium (MTT); the early apoptosis rates by staining with Annexin V and PI; the CD14 and CD11b expressions by flow cytometry (FCM); the promoter methylation status of hPer3 gene by MS-PCR; and the hPer3 mRNA expressions levels by RT-PCR.</p><p><b>RESULTS</b>The promoter methylation rates of hPer3 in newly diagnosed (ND) group, partial remission(PR) group, complete remission (CR) group, relapse (R) group and control group were 93.65% (59/63), 54.39% (31/57), 24.66% (18/73), 61.54% (8/13) and 0% (0/40), and the hPer3 mRNA expression levels were 0.19 ± 0.08, 6.28 ± 2.11, 52.76 ± 14.17, 8.18 ± 4.36, 75.03 ± 18.16, respectively. There was a significant statistic difference between any two group (P < 0.01) excepting for between PR and R group (P > 0.05). After DCA treatment, the promoter hypermethylation status of hPer3 was reduced and the mRNA and CD14, CD11b expression levels were up regulated in a dose dependent manner with an induction of cell apoptosis.</p><p><b>CONCLUSIONS</b>Promotor methylation status and mRNA expression of hPer3 gene may be indicators for evaluating AML. DCA can induce the expression of hPer3 gene and cells apoptosis in AML.</p>

Comparison of 99Tcm-DMSA renal cortical scintigraphy and ultrasonography in diagnosing acute pyelonephritis in children / 中华核医学杂志

Objective To compare the diagnostic value of renal ultrasound scan (RUS) and 99Tcmdimercaptosuccinic acid (DMSA) renal scintigraphy in children with acute pyelonephritis (APN). Methods In all, 165 children with initial clinical diagnosis of APN, aged from 1.5 months to 11 yrs ( median 20 months), were included in the study, all of which were examined with RUS and DMSA renal scientigraphy. The diagnosis with DMSA renal scientigraphy results was taken as the standard reference to evaluate the diagnostic sensitivity and specificity of RUS. Results Of 99 out of all 330 kidneys that were found abnormal on DMSA renal scientigraphy, 31 were abnormal on RUS. Of the rest normal kidneys on DMSA scans renal scientigraphy, 4 were abnormal on RUS. Thus diagnostic sensitivity of RUS for APN was 31.3%(31/99) and specificity was 98.3% (227/231). Conclusions Although RUS provides with high diagnostic specificity for children with APN, its low sensitivity may underestimate the clinical evaluation of APN.More often than not, 99Tcm-DMSA renal scientigraphy is a clinical necesscity for the definite RUS diagnosis.

The relationship between the genotype of hepatitis B virus and clinical and liver pathological features of infected patients in the Zhoushan Islands, China / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the genotypes of hepatitis B virus and the clinical and liver pathological features of patients with chronic hepatitis in the Zhoushan Islands.</p><p><b>METHODS</b>One hundred eighty HBV DNA positive chronic hepatitis patients with HBV markers were enrolled in this study. They were at least second generation Zhoushan Island residents. One hundred forty-seven of them were males and 33 were females with an average age of 39.0+/-11.3. Among the 180 patients, 17 had ASC, 57 had mild CHB, 48 moderate CHB, 9 severe CHB, 6 SHB, 39 LC, and 4 had HCC. The genotypes of their serum HBV were detected by using PCR integrated with Tagman MGB probe technology, and their serum HBV markers, HBV DNA and liver functions were also examined. Out of 180 patients, 129 accepted a liver biopsy. A pathological evaluation was then performed.</p><p><b>RESULTS</b>HBVs of genotype C, 135 cases (75.0%), of B, 40 cases (22.2%), and of B+C, 5 cases (2.8%) were found among these 180 patients. No genotype A or D HBV were found. The proportions of genotype C virus were 7/17, 86/114, 34/39, 6/6 in ASC, CHB, LC and SHB patients. In the hepatocellular carcinoma patients, there were 2 each of genotype B and C. Among the 99 patients with genotype C HBV, 84 cases (84.8%) showed moderate and severe inflammation histologically in their livers and among the 30 patients with B, 7 cases (23.3%) showed moderate to severe inflammation in their livers (z = 6.47, P less than 0.01). The proportion of genotype C HBV was significantly different from that of genotype B HBV in those that showed moderate and severe (S3-4) liver fibrosis. In patients infected with genotype C HBV who had moderate and severe liver pathological changes, their clinical manifestations reflected better the histological alterations of their livers.</p><p><b>CONCLUSION</b>Genotypes C, B and B+C HBV were found in CHB patients in the Zhoushan Islands of China, and type C was the predominant one. The liver pathological damage level of genotype C HBV infected patients is more serious than that of genotype B.</p>

An evaluation for the function and significance concerned to alterations of P16 3D structure with gene mutation in esophageal squamous cell carcinoma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathologic significance of P16 3D structure alterations in human esophageal squamous cell carcinoma(ESCC) so as to open a new approach for the research on clinical prevention and treatment of ESCC.</p><p><b>METHODS</b>All three techniques of polymerase chain reaction-single strand comformation polymorphism (PCR-SSCP), DNA sequencing and computerized three-dimensional protein-modeling were applied to analyze and determine the gene mutations and the computerized 3D changes of P16 protein molecule.</p><p><b>RESULTS</b>The p16 gene abnormality were detected from thirty-three cases of sixty-nine ESCC, among which twenty-six cases of ESCC showed the alterations of amino acid residues located within the P16 functional domains (classified as group M2), but other seven cases displayed the amino acid changes happened to beyond the domains and far from the p16-CDK4/6 binding site (defined as group M1). The statistical analysis revealed that the significant differences in lymph node metastasis, distance metastasis and stage of clinical were found between M2 and M1 groups (P<0.05). However no significant difference in sex, age, invasion depth of tumor was observed (P>0.05).</p><p><b>CONCLUSION</b>The mutations of p16 gene will alter the P16 protein structure. The four ankyrin repeats are the critical regions of P16 protein. The abnormalities in the ankyrin repeats will seriously alter the 3D structure and the activity of P16 protein, with resulting in lymph node metastasis, distance metastasis, and clinically advanced stage. The above results render an authentic criterion to the selection of the clinical cases with high risk of metastasis.</p>