BACKGROUND/OBJECTIVES: The study examined the association between homocysteine and diabetes mellitus in patients with H-type hypertension and assessed the possible effect modifiers. SUBJECTS/METHODS: This cross-sectional study included 1,255 eligible participants in the ‘H-type Hypertension Management and Stroke Prevention Strategic International Science and Technology Innovation Cooperation Project’ among rural Chinese people with H-type hypertension. A multivariate logistic regression model was used to evaluate the relationship between homocysteine and diabetes mellitus. RESULTS: The mean level of total homocysteine (tHcy) in the diabetes mellitus population was 19.37 μmol/L, which was significantly higher than the non-diabetic patients (18.18 μmol/L). When tHcy was analyzed as a continuous variable, the odds ratio (OR) of diabetes was 1.17 (95% confidence interval [CI], 1.01–1.35; per interquartile range). When tHcy was stratified according to the quintile, the ORs for diabetes were 2.86 (95% CI, 1.22–6.69) in the highest quintile (tHcy ≥ 20.60 μmol/L) compared to the reference group (tHcy < 12.04 μmol/L). When tHcy was grouped by 15 μmol/L and 20 μmol/L, patients with tHcy ≥ 20 μmol/L had a significantly (P = 0.037) higher risk of diabetes (OR, 2.03; 95% CI, 1.04–3.96) than in those with tHcy < 15 μmol/L. Subgroup analysis showed that the tHcy-diabetes association was unaffected by other variables. CONCLUSION In this study of rural Chinese people with H-type hypertension, the tHcy levels showed a positive association with diabetes mellitus. This independent association is unaffected by other potential risk factors.

Objective:To investigate the effect of silencing hepatocyte growth factor receptor (c-Met) expression on the biological characteristics of HCT116 colon cancer cells.Methods:Cellular model of c-Met transient transfection was established by using small interfering RNA (siRNA), the expression of c-Met in colon cancer cells was detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and western blot. The apoptosis assay, cell invasion assay, cell migration and other experiments were conducted to observe the effects of silencing c-Met on the biological characteristics of colon cancer cells.Results:RT-qPCR results showed that the relative expression levels of c-Met mRNA in siRNA-Met group, blank control group and siRNA negative control (siRNA-NC) group were 0.32±0.26, 1.01±0.03 and 1.05±0.23, respectively, and the difference was statistically significant ( P<0.05). Western blot analysis showed that the expression level of c-Met protein in the siRNA-Met group was 0.24±0.03, significantly lower than 1.23±0.06 in the blank control group and 1.18±0.11 in the siRNA-NC group ( P<0.05). The cell counting kit-8 (CCK8) results showed that the 72-hour absorbance (A) values of the siRNA-Met group, blank control group and the siRNA-NC group were 1.13±0.05, 1.48±0.08 and 1.53±0.07, respectively, and the difference was statistically significant ( P<0.01). Cell cycle results showed that the proportion of cells in G 2/M phase was (14.65±1.41)% in siRNA-Met group , (5.07±0.70)% in blank control group and (5.63±0.71)% in siRNA-NC group, and the difference was statistically significant ( P<0.05). The expression levels of cell cycle regulatory proteins Cdc25c and cyclin B1 in siRNA-Met group were significantly decreased. The apoptotic rate in siRNA-Met group was (5.85±0.35)%, significantly higher than (1.00±0.17)% in blank control group and (0.91±1.14)% in siRNA-NC group ( P<0.05). The expression level of apoptosis-related protein Bcl-2 in the siRNA-Met group was significantly decreased while Bcl-2 associated X protein (BAX) expression level was significantly increased. The cell scratching result showed that the cell migration abilities of the siRNA-Met group, blank control group and the siRNA-NC group were (51.33±8.62)%, (100.00±3.72)% and (102.33±6.43)%, respectively, and the difference was statistically significant ( P<0.05). The number of cell penetrating into the basement membrane of the siRNA-Met group, blank control group and the siRNA-NC group were 47.50±10.60, 100.00±5.33 and 102.50±10.61, respectively, and the difference was statistically significant ( P<0.05). The expressions of invasion related proteins including MMP-2 and MMP-9 in siRNA-Met group were decreased significantly. Conclusions:c-Met plays an important role in maintaining the biological characteristics of colon cancer cells. Inhibition of c-Met may have important values in the treatment of colon cancer.

Objective:To investigate the effect of silencing hepatocyte growth factor receptor (c-Met) expression on the biological characteristics of HCT116 colon cancer cells.Methods:Cellular model of c-Met transient transfection was established by using small interfering RNA (siRNA), the expression of c-Met in colon cancer cells was detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and western blot. The apoptosis assay, cell invasion assay, cell migration and other experiments were conducted to observe the effects of silencing c-Met on the biological characteristics of colon cancer cells.Results:RT-qPCR results showed that the relative expression levels of c-Met mRNA in siRNA-Met group, blank control group and siRNA negative control (siRNA-NC) group were 0.32±0.26, 1.01±0.03 and 1.05±0.23, respectively, and the difference was statistically significant ( P<0.05). Western blot analysis showed that the expression level of c-Met protein in the siRNA-Met group was 0.24±0.03, significantly lower than 1.23±0.06 in the blank control group and 1.18±0.11 in the siRNA-NC group ( P<0.05). The cell counting kit-8 (CCK8) results showed that the 72-hour absorbance (A) values of the siRNA-Met group, blank control group and the siRNA-NC group were 1.13±0.05, 1.48±0.08 and 1.53±0.07, respectively, and the difference was statistically significant ( P<0.01). Cell cycle results showed that the proportion of cells in G 2/M phase was (14.65±1.41)% in siRNA-Met group , (5.07±0.70)% in blank control group and (5.63±0.71)% in siRNA-NC group, and the difference was statistically significant ( P<0.05). The expression levels of cell cycle regulatory proteins Cdc25c and cyclin B1 in siRNA-Met group were significantly decreased. The apoptotic rate in siRNA-Met group was (5.85±0.35)%, significantly higher than (1.00±0.17)% in blank control group and (0.91±1.14)% in siRNA-NC group ( P<0.05). The expression level of apoptosis-related protein Bcl-2 in the siRNA-Met group was significantly decreased while Bcl-2 associated X protein (BAX) expression level was significantly increased. The cell scratching result showed that the cell migration abilities of the siRNA-Met group, blank control group and the siRNA-NC group were (51.33±8.62)%, (100.00±3.72)% and (102.33±6.43)%, respectively, and the difference was statistically significant ( P<0.05). The number of cell penetrating into the basement membrane of the siRNA-Met group, blank control group and the siRNA-NC group were 47.50±10.60, 100.00±5.33 and 102.50±10.61, respectively, and the difference was statistically significant ( P<0.05). The expressions of invasion related proteins including MMP-2 and MMP-9 in siRNA-Met group were decreased significantly. Conclusions:c-Met plays an important role in maintaining the biological characteristics of colon cancer cells. Inhibition of c-Met may have important values in the treatment of colon cancer.

Objective To investigate the diagnostic value of YKL‐40 and CEA in malignant pleural effusion .Methods There were 45 cases of benign pleural effusion and 52 patients with malignant pleural effusion in this study from November 2013 to No‐vember 2014 .Enzyme linked immunosorbent assay(ELISA) and electrochemi lumine scence immunoassay(ECLIA) method were carried out to detect the concentration of YKL‐40 and CEA respectively .The differences of the two groups of patients between YKL‐40 and CEA levels were compared ,and the correlation between YKL‐40 and clinical pathology of malignant pleural effusion were analyzed .In addition ,the receiver‐operating characteristic curve(ROC curve) was used to compare the diagnostic value be‐tween YKL‐40 and CEA .Results The average value of YKL‐40 in malignant pleural effusion was (189 .5 ± 147 .0)ng/mL ,and sig‐nificantly higher than in benign pleural effusion group(P< 0 .05) .The elevated level of YKL‐40 was related to tumor types and lymphatic metastasis(P<0 .05) ,but it had no correlation with the gender ,age and degree of tumor differentiation(P>0 .05) .The diagnostic sensitivity and specificity of YKL‐40 was 80 .9% and 51 .2% ,which was lower than CEA(83 .1% and 74 .6% )(P<0 .05) .However ,the sensitivity and specificity was 90 .6% and 88 .2% when combined the two biomarkers together .Conclusion YKL‐40 have a certain clinical diagnostic value in malignant pleural effusion ,it indicate to adenocarcinoma or advanced cancer when the level of YKL‐40 rised .Since the sensitivity and specificity is lower than traditional biomarker of CEA ,we should combine with the other tumor markers to improve the diagnostic accuracy .

OBJECTIVETo establish a one-step four multiplex reverse transcription polymerase chain reaction (RT-PCR) method based on Homo-Tag Assisted Non-Dimer System (HAND) system for simultaneous detection of 4 diarrhea viruses of rotavirus, astrovirus, norovirus and sapovirus.

METHODSPrimers were designed according to the conserved genome sequence of the 4 viruses and the homologous tail sequences were added to the 5' end. The multiplex RT-PCR system was constructed by optimizing the PCR parameters such as the concentration of universal tag primer and genome-specific Homo-Tailed primers. The specificity, stability and sensitivity of the method were evaluated systematically.

RESULTSThe 4 multiplex RT-PCR methods based on HAND system was established successfully. Specificity analysis showed no cross reaction between the 4 diarrhea viruses. The sensitivity analysis showed detection limits for rotavirus, astrovirus, norovirus and sapovirus of 48, 1.92, 9.6 and 48 pg per reaction, respectively.

CONCLUSIONThe established HAND system-based multiplex RT-PCR assay allows simple, rapid, specific, sensitive, and stable for detection of the 4 common diarrhea viruses at low costs and is suitable for application in general medical laboratories.

Astroviridae ; genetics ; isolation & purification ; Diarrhea ; virology ; Feces ; virology ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Norovirus ; genetics ; isolation & purification ; RNA, Viral ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Rotavirus ; genetics ; isolation & purification ; Sapovirus ; genetics ; isolation & purification

[Objective] To learn about the genetic diversity,we studied the five X-chromosomal STR (X-STR) loci in Guangdong Han Nationality Groups.[Methods] The five Loci (DXS6803,DXS981,DXS6809,DXS6789,and DXS7132) were amplified in a pentaplex PCR reaction.PCR products were analyzed using capillary electrophoresis and ABI prism 3100 Genetic Analyzer,with GeneMapper ID 3.1 Analysis Software.[Results] A total of 363 individuals (181 unrelated male and 182 unrelated female) from Guangdong Han population were tested,54 alleles were observed for these loci.Polymorphism information content is 0.6935 ～ 0.8177.Power of discrimination in females was 0.8976 ～ 0.9562.Mean exclusion chance for X-STR in standard trios with daughters was 0.7805 ～ 0.8467.[Conclusion] The five loci in the multiplex system provide high polymorphism information for forensic identification and paternity testing,particularly for difficult paternity deficiency cases.

OBJECTIVETo establish a nine X-chromosome short tandem repeats (X-STR) loci multiplex PCR method and study the polymorphism of the 9 X-STR loci,and to determine its application in kinship tests for forensic medicine.

METHODSA fluorescent multiplex PCR that simultaneously amplifies 9 X-STR loci, i.e. DXS7133, DXS981, DXS7424, DXS6789, DXS7132, GATA165B12, DXS101, GATA31E08 and DXS10011, were set up. PCR products were analyzed using capillary electrophoresis and ABI prism 3100 Genetic Analyzer with GeneMapper ID 3.1 analysis software.

RESULTSWhen 251 unrelated male and 112 unrelated female individuals from southern China were tested, 111 alleles were detected. The power of discrimination in females was 0.5837-0.9959. Mean exclusion chance for X-STR in standard trios with daughters was 0.4072-0.9511. Polymorphism information content was 0.4481-0.9531.

CONCLUSIONThe results demonstrate that the 9 loci in the multiplex system provide high polymorphism information, and the multiplex system provides a fast technology for forensic identification and paternity testing. The X-STR multiplex system can complement the analysis of AS-STR and Y-STR efficiently.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Chromosomes, Human, X ; genetics ; Female ; Humans ; Male ; Microsatellite Repeats ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic

This article review the advance in interpretation of mixed forensic stains using DNA profiling, including autosome STR profiling, sex profiling determined by PCR, Y-specific STR profiling, mitochondrial DNA profiling and single nucleotide polymorphism profiling. The statistics methods for mixed stain has also been reviewed.
Alleles ; Blood Stains ; Chromosomes, Human, Y/genetics* ; DNA/genetics* ; DNA Fingerprinting/methods* ; Female ; Forensic Medicine/methods* ; Humans ; Male ; Polymerase Chain Reaction ; Tandem Repeat Sequences

Objective To study the expression of three survivin splicing variants in gastric cancer and normal gastric mucosa and to evaluate relationship among the survivin variants' expression and proliferation, apoptosis in gastric cancer. Methods Real time quantitative RT-PCR was used to analyze survivin variants expression in 77 paired tumors and normal gastric mucosa in frozen samples at the mRNA level. The cell proliferation and apoptosis were measured by Ki-67 immunoln's to chemical analysis and TUNEL method in paraffin-embedded block of same cases, respectively. Results The sarvivin splicing variants were remarkably up-regulated in gastic cancers compared with those in normal tissues (P

Objective To observe the degeneration of Brugia malayi in Meriones unguiculatus model. Methods Microfilaria of Brugia malayi derived from Meriones unguiculatus was used to infect Anopheles sinensis . Infective stage larvae (L 3) from mosquito vector were collected and inoculated into abdomen of Meriones unguiculatus. Successive 33 generations of the parasite in the rodent model have been observed. Results Since 1974 when the animal model was established, the parasite has lasted for 33 generations, the positive rate of Meriones unguiculatus with microfilaria gradually reduced from 80% of the 28th generation to 16% of the 32nd generation and finally to 0 at the 33rd generation. Conclusion It became difficult for the larvae of Brugia malayi to develop and/or reproduce in the animal model after multiple inoculations for generations.