OBJECTIVETo explore the influence of the DNA repair gene ERCC2 single nucleotide polymorphisms (SNPs) rs13181, rs1618536, and rs1799793 on male idiopathic infertility in Ningxia, China.

METHODSUsing MassArray, we conducted a case-control study and genotyped three ERCC2 SNPs rs13181, rs1618536, and rs1799793 for 351 males (aged 31.0 +/- 4.2 years) with idiopathic infertility and another 327 normal fertile men (aged 33.0 +/- 5.9 years) as controls.

RESULTSThe ERCC2 AnyG-anyA-anyA genotypes were significantly associated with an increased risk of idiopathic infertility (OR 0.414, 95% CI 0.176 - 0.970), while the three single ERCC2 SNPs rs13181, rs1618536, and rs1799793 showed no significant differences between the cases and controls (P > 0.05).

CONCLUSIONThe ERCC2 SNPs rs13181, rs1618536, and rs1799793 play a role of interaction in male idiopathic infertility in Ningxia, contributing to the risk of the disease.

Adult ; Case-Control Studies ; China ; DNA Repair ; Genotype ; Humans ; Infertility, Male ; genetics ; Male ; Polymorphism, Single Nucleotide ; Xeroderma Pigmentosum Group D Protein ; genetics

OBJECTIVETo test the dissolution rate of silymarin dropping pill as well as to be compared with other three commercial products of the silymarin.

METHODBy UV spectrophotometry, we studied the dissolution conditions of silymarin dropping pill and compared its dissolution rate with Yiganling tablets (film-coating, sugar-coating) and Legalon capsule which are available in the market.

RESULTThe dissolution parameters T50 and Td of silymarin dropping pill, Yiganling tablet (film-coating), Yiganling tablet (sugar-coating) and Legalon capsule are 6.78, 9.85 min, 51.01, 73.78 min, 74.35, 86.97 min and 53.10, 72.65 min.

CONCLUSIONThe dissolution rate of silymarin dropping pill is superior to that of two kinds of Yiganling tablets and Legalon capsule.

Capsules ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Silymarin ; administration & dosage ; chemistry ; Solubility ; Spectrophotometry, Ultraviolet ; Tablets

Objective To detect the expression levels of high mobility group box chromosomal protein 1 (HMGB1) and Th17 cells transcription factors, related cytokines in peripheral blood of rheumatoid arthritis (RA) patients and analyze the relations between HMGB1 and CRP, ESR, RF in RA patients. The other aim of this study is to identify the expression level of HMGBI and the relationship between HMGB1 and Th17 in RA patients. Methods The mRNA levels of HMGB1, RORyt, interleukin (IL)-17 in the peripheral blood mononuclear cells (PBMC) were determined by quantitative real-time PCR (QRT-PCR) from 80 patients with rheumatoid arthritis,including 32 RA patients in stable phase and 48 patients in active phase, and 50 healthy volunteers. The concentration of HMGB1, IL-23, IL-17 in plasma were detected by enzyme linked immunosorbent assay (ELISA), one-way ANOVA and Spearman's correleation were adopted for statistical analysis.Results The mRNAs of HMGBI, RORyt and IL-17 in RA patients were higher than that in healthy control group (P<0.05), especially in active RA patients [ HMGB 1 (0.424±0.262) pg/ml, RORγt (0.34±0.25) pg/ml,IL-17 (1.42±0.38) pg/ml,P<0.01 ] when compared with patients with stable disease. The concentration of HMGB1, IL-23 and IL-17 in the plasma of RA patients was higher than that of the healthy control group (P< 0.05), and was positively correlated with the expression levels of HMGB1, Th 17-associated factors and the level of CRP, ESR, RF in RA patients' plasma(P<0.05). Conclusion The HMGB1 and Thl7 cells levels are higher in active RA patients than those in patients with stable disease, arid there is significant positive correlation between them. Detection of peripheral HMGB1 and Thl7 cell-specific transcription factors or related cytokines can help to understand the development and progress of rheumatoid arthritis and provide clues for new treatment targets for RA.

Discovery of the RNA interference (RNAi) pathway has led to exciting new strategies for developing HIV treatment. This study was to find out the highly effective and conserved siRNA target sequences for improving RNAi-based therapy against the HIV-1. We constructed 30 shRNA expression plasmids for expressing different siRNAs targeted to HIV-1 vif and co-transfected them with the pNL4-3 to score for its ability to inhibit the expression of p24 protein of HIV-1. Then, the highly effective siRNAs targeting sequences were selected to align with 625 HIV-1 sequences in database including all HIV-1 subtypes to ana lyze their conserved character. In addition, vif37 the highly effective and most conserved target sequence was confirmed of its sequence-specific inhibition by independent reporter assays. MT-4 cell transduced with lentiviral shRNA-vif37 vector could inhibit HIV-1(NL4.3) replication in vitro. Moreover, MT-4-vif37 cloned from transduced MT-4 cell could stably express shRNA-vif37 and inhibit virus replication more efficiently when challenged with high titer virus. These results showed that RNAi has great potential as an antiviral gene therapy approach and supports the efforts to develop treatment for HIV-1-infected individuals.
Acquired Immunodeficiency Syndrome ; therapy ; Base Sequence ; Humans ; Lentivirus ; genetics ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; chemistry ; genetics ; Virus Replication ; vif Gene Products, Human Immunodeficiency Virus ; antagonists & inhibitors ; genetics

OBJECTIVESTo review the preliminary clinical experience with high-field-strength intraoperative magnetic resonance imaging (iMRI) suite with neuronavigation system in the pituitary adenoma operation with transsphenoidal approach.

METHODSFrom March 2009 to December 2010, 31 patients [range, 29 - 76 years, mean age (47 ± 11) years]of pituitary adenoma were operated with transsphenoidal approach and intraoperatively with a movable 1.5 T high-field-strength iMRI suite in combination with neuronavigation system. Tumor size was 1.8 - 7.3 cm, mean (3.5 ± 1.2) cm. Twenty-five cases were non-functional pituitary adenoma, 4 cases were prolactin-secreting pituitary adenoma, 2 cases were growth hormone-secreting pituitary adenoma. Thirty patients' resection with transnasal transsphenoidal approach were performed, one patient with transoral transsphenoidal approach was performed.

RESULTSIn 12 cases of 30 patients who planed to totally remove tumor, iMRI had revealed residual lesions and resulted in the change of the surgical strategy, 2 invasive cavernous sinus cases no further resection of the tumor because of internal carotid artery encasement, the other 10 cases resected further, eventually. Finally, 8 cases were totally removed. The ratio of total removal tumor was enhanced to 86.7% (26/30) from 60.0% (18/30). There was no perioperative mortality.

CONCLUSIONSHigh-field-strength iMRI suite with neuronavigation system provides valuable information of tumor resection that allows intraoperative modification of the surgical strategy. It could be very helpful to maximize the resection of the pituitary adenoma and minimize the injury to neurological function.

Adenoma ; surgery ; Adult ; Aged ; Cavernous Sinus ; surgery ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Monitoring, Intraoperative ; methods ; Neuronavigation ; methods ; Pituitary Neoplasms ; surgery

In order to fabricate the HLA-DQA1 genotyping chip and develop an integrated, parallel technical platform to type HLA system, a pair of primers and a set of probes were designed according to the sequences of HLA-DQA1 exon 2, where the polymorphism is concentrated. The oligonucleotide chip was made with the methods developed in our laboratory. The target DNA was asymmetrically amplified with the labeled sense primer. The signals were scanned and analyzed after the hybridization between microarray and PCR product. The allele types of the samples were identified. The result was verified by the standard DNA and DNA sequencing. The results showed that the genotyping was successfully carried out in 50 standard DNA samples and 50 clinical samples. Among them, results of the 50 standard DNA samples matched their templates. In the other 50 samples, results of the randomly selected 10 matched their sequencing results except that two of them got the incompletely result. In reproducible tests, the signal reappear rate was 95%. It is concluded that HLA-DQA1 genotyping by using our array system is simple and convenient with satisfied accuracy and reproducibility.
Genotype ; HLA-DQ Antigens ; genetics ; immunology ; HLA-DQ alpha-Chains ; Humans ; Oligonucleotide Array Sequence Analysis ; Oligonucleotide Probes ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the pathological feature of primary hepatic carcinoma and the clinical significance.

METHODSFrom August 2000 to December 2007, there were 89 patients with cirrhosis and carcinoma of liver who accepted whole liver resection. The whole liver was cut into 10 mm slices to examine the tumor size, number, distribution, capsule, satellite nodes, portal vein tumor thrombi (PVTT). The invaded adjacent tissue and lymph nodes were recorded, the distance from satellite to major tumor was measured, then histological examinations were carried out, and the final diagnosis was made by pathologists.

RESULTSThe total of 89 cases included hepatocellular carcinoma in 86 cases and cholangiocarcinoma in 3 cases; 53 cases with multiple tumors and 36 cases with solitary tumor; complete capsule only in 14 cases, no obvious margin in 11 cases, 13 cases had a major tumor in the right lobe and a small tumor in the left lobe; 8 of 25 cases with gross invaded tissue were confirmed by histological examination, 7 of 16 cases with swollen lymph nodes were infiltrated by cancer cells. There were 47 cases with PVTT (47.2%) and 39 cases with satellite nodes (43.8%). PVTT and satellite nodes increased with the increase of sizes and the numbers of the tumors. The distance from satellite node to major tumor mostly were 0.5 - 3.0 cm.

CONCLUSIONSThe whole explanted liver can completely reflect the characteristics of growth and infiltration of hepatic carcinoma. Attention must be paid to the small cancer lesions in another lobe, distal satellite nodes from major tumor, and tumor thrombi in a small branch of portal vein, which can not be found by imaging, and might influence the curative effectiveness after liver resection or transplantation.

Adult ; Carcinoma, Hepatocellular ; pathology ; surgery ; Female ; Hepatectomy ; Humans ; Liver ; pathology ; Liver Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Young Adult

AIM To study the effects of Yizhi Congming Decoction (Ginseng Radix et Rhizoma,Poria,Polygoni multiflori Radix praeparata,etc.) on learning and memory deficits through reducing tau hyperphosphorylation in Aβ-induced AD mice and its mechanism of action.METHODS Sixty ICR mice were randomly and equally assigned into six groups.Sham group (injection of 3 μL normal saline into left hippocampus),model group,donepezil group and each group of Yizhi Congming Decoction (injection of 3 μL Aβ25-35 into left hippocampus).Donepezil group and three treatment groups were gavaged with donepezil and Yizhi Congming Decoction.After 15 days of administration,immunohistochemistry was used to observe the expressions of Ser404 and Thr231.Western blot was performed to evaluate the levels of phosphatidylinositol-3-kinase (PI3K),threonine/serine protein kinase B (AKT) and glycogen synthase kinase 3β (GSK3β).RESULTS Yizhi Congming Decoction attenuated the levels of phosphorylated tau at the Thr231 and Ser404 sites in the hippocampus and increased the phosphorylation levels of PI3K,AKT and GSK3β.CONCLUSION Yizhi Congming Decoction effectively provides protection against learning and memory deficits and inhibits hyperphosphorylated tau protein expression in the hippocampus.The possible mechanism may occur via the PI3 K/Akt-dependent GSK3 β signalling pathway.

Objective To observe protective effects on rat serum cardiac enzymes and the antioxidant capacity of selenium and vitamin E.Methods According to body weight and 2 × 2 factorial design,eighty male Wistas rats were randomly divided into four groups:low selenium and low vitamin E group(feed containing 23.42％ of the low selenium yeast,excluding vitamin E),low selenium and adequate vitamin E group (feed containing 23.42％ of the low selenium yeast and vitamin E 160 mg/kg),adequate selenium and low vitamin E group(feed containing 46.84％ of the low selenium yeast and sodium seleni 0.25 mg/L in water,excluding vitamin E),adequate selenium and adequate vitamin E group(feed containing 46.84％ of the low selenium yeast,vitamin E 160 mg/kg and sodium selenite 0.25 mg/L in water),20 rats every group.Rats were feed with synthetic feed,and given intraperitoneal anesthesia after 26 weeks of feeding.Blood was collected to observe the impact of selenium and vitamin E on rat cardiac enzymes and myocardial antioxidant capacity and their interactions.Serum creatine kinase (CK) was measured using the continuous monitoring method,creatine kinase isozymes (CK-MB) and lactate dehydrogenase(LDH ) using the immune suppression method,the whole blood GSH-Px assay using the dithiobis nitrohenzoic acid(DTNB) method,serum superoxide dismutase(SOD) using the xanthine oxidase method,total antioxidant capacity (T-AOC) using the complex colorimetry method,the content of propylene glycol (MDA) using the thiobarbituric acid colorimetric method,and reactive oxygen species(ROS) using the colorimetric method.Results Group differences of serum CK,CK-MB,LDH,whole blood GSH-Px activity,serum T-AOC vitality,MDA and ROS content were statistically significant(F=9.797,17.041,48.399,3.744,224.900,49.384,5.045,all P＜ 0.05).Compared with the two low selenium groups and one adequate selenium group,the vitalities of CK,CK-MB,LDH and the contents of MDA[(1577.75 ± 451.87),(1239.15 ± 344.99),(884.25 ± 133.84)U/L,(5.688 ±1.169) × 103 nmol/L; (1474.21 ± 398.38),(1014.84 ± 215.40),(523.00 ± 98.05)U/L,(4.035 ± 0.487 ) × 103 nmol/L and (1180.10 ± 245.51),(948.75 ± 173.68),(676.70 ± 193.63)U/L,(3.406 ± 0.146) × 103 nmol/L]increased significantly in adequate selenium and adequate vitamin E group[( 1056.80 ± 250.98),(721.70 ±129.98),(404.65 ± 72.49)U/L,(3.010 ± 1.270) × 103 nmol/L,all P ＜ 0.05) ].The activity of GSH-Px was obviously increased in the two adequate selenium groups[ (96.611 ± 8.238) × 103,(103.024 ± 8.217) × 103 U/L,all P ＜ 0.05],compared with the two low selenium groups[ (60.356 ± 8.179) × 103,(63.117 ± 8.281) × 103 U/L].Selenium affected the activities of CK,CK-MB and LDH(F =27.09,31.58,29.66,all P＜ 0.01 ),and vitamin E affected the activities of CK-MB and LDH(F=18.9,11.2.all P＜ 0.01 ),but both selenium and vitamin E had no interactions on the activities of CK,CK-MB and LDH (F=0.02,0.001,2.22,all P＞0.05).Selenium affected the activity of GSH-Px and the content of MDA(F=6.74,95.68,all P＜ 0.05),vitamin E affected the activity of T-AOC,the contents of MDA and ROS(F=6.42,36.73,8.43,all P＜0.05),but selenium and vitamin E had interactions only on the content of MDA(F =13.82,P＜ 0.05).Conclusions Long-term selenium or vitamin E deficiency,can reduce the body's antioxidant capacity,leading to the occurrence of myocardial injury.Selenium and vitamin E can improve the body's oxidation capacity,playing a role in myocardial protection.

Objective To study the effects of selenium(Se) and protein on cardiac morphology and expression of cellular glutathione peroxidase(GPX1 ) and mitochondrial thioredoxin reductase(TR2) in rat myocardial tissue.MethodsSixty healthy weaning male Wistar rats were randomly divided into four groups by two factors two levels factorial design(n =15).Drinking water was divided into two levels of Se-deficient(0 mg/L) and Se-adequate (0.25 mg/L); diet was divided into two levels of protein-deficeient (10％ protein and 0.008 mg/kg Se) and protein-adequate(20％ protein and 0.015 - 0.026 mg/kg Se).The rats were killed after feeding for one year.Pathological changes in myocardial tissues were observed under light microscope.The expression of GPX1 and TR2 in rat myocardial tissue was detected by immunohistochemistry and Western blotting.Results Compared between groups,the difference of the rate of myocardial necrosis in rats was statistically significant(x2 =11.04,P ＜ 0.05),in which Se-deficient protein-deficient group [66.7％ (8/12) ] was significantly higher than Se-adequate proteinadequate group [ 7.1％ ( 1 / 14),x2 - 11.06,P ＜ 0.05 ].GPX 1 positive rates in Se-deficient protein-deficient group,Se-adequate protein-deficient group,Se-deficient protein-adequate group and Se-adequate protein-adequate group were 0(0/12),81.8％(9/11 ),10.0％(1/10) and 100.0％(14/14),respectively,in rat myocardial tissue determined by immunohistochemistry.Of which,Se-adequate protein-deficient group and Se-adequate protein-adequate group were significantly higher than Se-deficient protein-deficient group and Se-deficient protein-adequate group(x2 =12.88,8.14 and 35.89,32.60,all P ＜ 0.05).The positive expression rates of TR2 in rats myocardial tissue of the four groups were 0(0/12),81.8％(9/11),0(0/10) and 100.0％(14/14),respectively.Of which,Se-adequate proteindeficient group and Se-adequate protein-adequate group were significantly higher than Se-deficient protein-deficient group and Se-deficient protein-adequate group (x2 =28.67,18.25 and 35.89,32.60,all P ＜ 0.05).The four groups'results of the overall mean of the relatively value of protein expression of GPX1 in cardiac tissue by Western blotting were 0.87 ± 0.13,1.18 ± 0.13,0.95 ± 0.13 and 1.74 ± 0.23,respectively.Through analysis of variance of factorial design,the effects of Se and protein on protein expression of GPX1 in the heart were statistically significant(F=124.93,43.16,all P＜ 0.05).And there was interaction between them(F=24.10,P＜ 0.05).The four groups'results of the overall mean of the relatively value of protein expression of TR2 in cardiac tissue by Western blotting were 0.63 ± 0.19,0.97 ± 0.24,0.55 ± 0.08 and 1.03 ± 0.31,respectively.Through analysis of variance of factorial design,the effect of Se on expression of TR2 in the heart was statistically significant(F =36.97,P ＜ 0.05).Conclusions Adequate Se and protein diet can increase the levels of GPX1 and TR2 in the heart compared to deficient Se and protein diet,can enhance anti-oxidizing ability,protect the myocardial endothelial cells,reduce degree of myocardial injury,and the combined effects of both are better.