Objective To screen the fragilex mental retardation 1 (FMR1) gene mutations and explore the frequency of FMR1 gene mutation in the population with mental retardation in South China.Methods Seventy-two patients (65 males and 7 females) with suspected fragile X syndrome (FXS) in South China were enrolled in our hospitals from October 2009 to April 2014.The CGG trinucleotide repeats in 5'UTR of FMR1 gene and CCG trinucleotide repeats in FMR2 gene were screened respectively by PCR.Southern blotting and capillary electrophoresis sequencing were performed in male patients without normal target bands and suspected female patients;patients with normal CGG alleles were,then,performed exons and 3'-UTR ofFMR1 gene amplification and sequencing.The frequency of FMR1 gene mutation in patients with mental retardation in different countries and regions was compared with statistical analysis.Results Six pedigrees with full mutation (one female and five males being the probands),one pedigree (mother and son) with FMR1 gene deletion and one pedigree (mother and son) with mutation in the transition region were identified in 72 patients with mental retardation.The prevalence of total mutation was 9.7％ (7/72) and that in male patients was 9.2％ (6/65).These results showed significant differences in prevalence as compared with the results from different countries and areas (P＜0.05);there were no variations in 3'UTR ofFMR1 gene and FMR2 gene mutation in the patients with FXS-like phenotype.Conclusions FMR1 mutation frequency may be higher in mental retardation population in southem China as compared with that in developed countries or areas.Targeted screening on the unexplained mental retardation pedigrees (family history) can improve the diagnosis of FXS.Importantly,deletion mutations screening should also be performed in suspected FXS subjects with normal CGG repeats.

OBJECTIVETo study the leptin resistance mechanism of Xiaoyan Decoction (XD) in lung cancer cachexia (LCC) rats.

METHODSAn LCC rat model was established. Totally 40 rats were randomly divided into the normal control group, the LCC model group, the XD group, and the positive control group, 10 in each group. After LCC model was set up, rats in the LCC model group were administered with normal saline, 2 mL each time. Rats in the XD group were administered with XD at the daily dose of 2 mL. Those in the positive control group were administered with Medroxyprogesterone Acetate suspension (20 mg/kg) by gastrogavage at the daily dose of 2 mL. All medication lasted for 14 days. The general condition and tumor growth were observed. Serum levels of leptin and leptin receptor in the hypothalamus were detected using enzyme-linked immunosorbent assay. Contents of neuropeptide Y (NPY) and anorexia for genomic POMC were detected using real-time PCR technique.

RESULTSSerum leptin levels were lower in the LCC model group than in the normal control group with statistical significance (P < 0.05). Compared with the LCC model groups, serum leptin levels significantly increased in the XD group (P < 0.01). Leptin receptor levels in the hypothalamus increased significantly in the LCC model group (P < 0.01). Increased receptor levels in the LCC model group indicated that either XD or Medroxyprogesterone Acetate could effectively reduce levels of leptin receptor with statistical significance (P < 0.01). There was also statistical difference between the XD group and the positive control group (P < 0.05). Contents of NPY was higher in the LCC model group than in the other groups with statistical difference (P < 0.05). There was no statistical difference in NPY between the normal control group and the rest 2 treatment groups (P > 0.05). There was statistical difference in POMC between the normal control group and the LCC model group (P < 0.05). POMC could be decreased in the XD group and the positive control group with statistical significance (P < 0.05), and it was more obviously decreased in the XD group (P < 0.05).

CONCLUSIONSLeptin resistance existed in LCC rats. XD could increase serum leptin levels and reduce leptin receptor levels in the hypothalamus. LCC could be improved by elevating NPY contents in the hypothalamus and reducing POMC contents, promoting the appetite, and increasing food intake from the periphery pathway and the central pathway.

Animals ; Cachexia ; drug therapy ; etiology ; Drugs, Chinese Herbal ; therapeutic use ; Eating ; Humans ; Hypothalamus ; metabolism ; Leptin ; metabolism ; Lung Neoplasms ; complications ; Neuropeptide Y ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate histopathologic examination of the testis tissue from testicular sperm aspiration (TESA).

METHODSWe analyzed the results of inverted microscopy and histopathologic examination of 96 samples of testis tissue from TESA, and compared the accuracy of the two methods in detecting sperm in the testis tissue.

RESULTSAmong the 11 cases in which sperm was found by inverted microscopy, 9 were confirmed by histopathologic examination, and among the 57 cases in which sperm was not detected by inverted microscopy, 11 (19.3%) were found with sperm by histopathologic examination. Histopathologically, the cases in which sperm was not found by inverted microscopy included Sertoli-cell-only syndrome (n = 34), maturation arrest (n = 12) and hypospermatogenesis (n = 11).

CONCLUSIONHistopathologic examination may reveal sperm in the TESA testis tissue proved to be sperm-absent by microscopy, and thus offer valuable information for a second testicular sperm retrieval.

Adult ; Azoospermia ; pathology ; Humans ; Male ; Middle Aged ; Sperm Count ; Sperm Retrieval ; Testis ; pathology ; Young Adult

OBJECTIVETo establish an animal model of congenital heart defect with decreased pulmonary blood flow for better understanding the pathophysiology of pulmonary vascular development and related regulatory mechanisms of congenital heart defect with decreased pulmonary blood flow.

METHODOne to two months old pigs were randomly divided into three groups: control group (group C, n = 6) with right chest small incisions induced transient pulmonary blood reduction; light-moderate stenosis groups (group T(1), n = 7): artificial atrial septum defect (ASD) plus controlled pulmonary artery banding to generate a systolic pressure gradient of 20 - 30 mm Hg (1 mm Hg = 0.133 kPa); severe stenosis groups (group T(2), n = 7): similar surgical procedures as group T(1), and controlled pulmonary artery banding to generate a systolic pressure gradient ≥ 30 - 50 mm Hg. 64-slice computed tomography scanning was performed at one month post operation. Arterial blood gas analysis, hemoglobin value, pulmonary vessel, ASD and banding ring diameters and trans-pulmonary artery banding pressure (Trans-PABP) were determined at two months post operation.

RESULTSOne pig died due to tracheal intubation accident in the C group, one pig died due to bowel obstruction in the T(1) group and two pigs died due to acute right heart failure and chronic heart failure respectively in T(2) group. 64-slice CT angiography results showed that aortic diameter of T(1) group was significantly lower than that of C group and banding diameter was significantly lower than aortic diameter in the T(1) and T(2) groups at one month post operation. Two months after operation, the size of ASD were (8.0 ± 0.5) mm and (8.9 ± 1.4) mm (P > 0.05) respectively in the T(1) and T(2) groups after operation. The Trans-PABP was significantly higher in the T(1) and T(2) groups than in C group (P < 0.01), and the Trans-PABP was significantly higher in the T(2) group than in T(1) group (P < 0.01). PaO2 and SaO2 in the T(1) and T(2) groups were significantly lower than those in C group.

CONCLUSIONArtificial atrial septum defect combined pulmonary artery banding procedures could be successfully used to establish model of congenital heart defect with decreased pulmonary blood flow and this model could help to understand the pathophysiology and monitor therapy efficacy for patients with congenital heart defect with decreased pulmonary blood flow.

Animals ; Disease Models, Animal ; Heart Defects, Congenital ; physiopathology ; Lung ; blood supply ; Pulmonary Artery ; physiopathology ; Pulmonary Circulation ; Pulmonary Veins ; physiopathology ; Swine

An improved everted gut sac method was applied to the study of prescription compatibility effect on the major components in Danshen extracts. With the separation and detection by HPLC-ECD, 5 major peaks could be detected in intestinal absorbed solution after prescription administration. Following the identification by HPLC-MS/MS, peak 2, 3, 4, and 5 were rosmaric acid, lithospermic acid, salvianolic acid B, and salvianolic acid A, respectively, which also confirmed with reference standards of those components. Through paralleling substance identification, peak 2, 3, 4, and 5 could be found as the major components in Danshen extracts, except Salvianolic acid E which is undetectable in intestinal solution. The contents of peak 2, 3, and 4 did not show difference before and after compatible prescription administrated, where the peak 5 had a significant increase in the same process. Those results revealed that peak 5, salvianolic acid A, might lead to an increasing pharmacological effect after prescription compatibility.
Animals ; Benzofurans ; analysis ; pharmacokinetics ; Caffeic Acids ; analysis ; pharmacokinetics ; Chromatography, High Pressure Liquid ; methods ; Depsides ; analysis ; pharmacokinetics ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacokinetics ; In Vitro Techniques ; Intestinal Absorption ; Lactates ; analysis ; pharmacokinetics ; Male ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Salvia miltiorrhiza ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

Objective To evaluate the clinical efficacy of radical surgery combined with irradiation in the treatment of rectal cancer and its effect on the angiogenesis and survival rate.Methods A total of 200 colorectal cancer patients admitted to Zhengzhou Central Hospital from March,2014 to March,2015 were recruited and divided into the observation group (n=105) and control group (n=95) by using random number table method.In the control group,radical surgery was performed,and radical surgery combined with irradiation was conducted in the observation group.The clinical efficacy,the serum levels of vascular endothelial growth factor-C (VEGF-C) and prostaglandin E2(PGE2) were statistically compared between the control and observation groups.The changes of the microvascular morphology and microvessel density (MVD) in the rectal cancer tissues were observed and recorded.The 3-year survival rate was calculated during postoperative follow-up.Results After corresponding treatment,the clinical overall response rate was 86.67％ in the observation group,and 70.53％ in the control group (P＞0.05).The 2-year survival rate did not significantly differ between two groups (P＞0.05).The 3-year survival rate in the observation group was significantly higher than that in the control group (P＜0.05).After treatment,the serum levels of VEGF-C and PGE2 were significantly improved in two groups (both P＜0.05).In the observation group,the serum levels of VEGF-C and PGE2 were significantly lower compared with those in the control group (both P＜0.05).The microvessel morphology in the cancer tissues remarkably differed between two groups.The microvessel diameter did not significantly differ,whereas the lumen diameter in the observation group was significantly smaller than that in the control group.The MVD in the observation group was 12.25±3.35,significantly lower than 28.14± 17.26 in the control group (P＜0.05).Conclusion Radical surgery combined with irradiation is an efficacious treatment of rectal cancer,which can effectively improve the serum levels of VEGF-C and PGE2,decrease the MVD,reduce the lumen diameter in the cancer tissues,lower the angiogenesis in rectal cancer and enhance the survival rate,which deserves widespread application in clinical practice.

Constriction, Pathologic ; Fibrosis ; Humans ; Lung ; Mediastinitis/therapy* ; Sclerosis

OBJECTIVETo detect the expression and variation of p53 gene during tree shrews' hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B1 (AFB1).

METHODSTree shrews were divided into four groups: the tree shrews were infected with HBV and fed with AFB1 in group A, only infected with HBV in group B, fed with AFB1 alone in group C, and normal control in group D. All the tree shrews were performed liver biopsy every 15 weeks. The tissues of liver and tumor were detected by immunohistochemistry and molecular biotechnologies.

RESULTS(1) The incidence of hepatocellular carcinoma (HCC) in group A (66.7%) was higher than that in Group B and C (30%). HCC appearance in group A was earlier than that in group C (120.0 weeks +/-16.6 weeks vs 153.3 weeks +/-5.8 weeks, t = 3.336, P<0.01). (2) Mutated p53 protein was not found before the 75th week of the experiment in each group. (3) At the 105th week, the expression rates of mutated p53 protein were 78.6%, 60% and 71.4% in group A, B and C respectively, which were much higher than that (10%) in group D (x2 > or = 5.03, P<0.05). An abnormal band of p53 gene was detected in both group A and C. (4) The mutation points of p53 gene in liver cancer of tree shrew were at codon 275, 78 and 13. The nucleotide sequence and amino acids sequence of tree shrew's wild-type p53 showed 91.7% and 93.4% homology with those of human p53 respectively.

CONCLUSIONSThere is a remarkable synergistic effect between HBV and AFB1 on HCC. Mutated p53 protein is expressed before HCC occurrence, which promotes the development and progress of HCC. HBV and AFB1 may synergistically induce p53 gene mutation.

Aflatoxin B1 ; toxicity ; Animals ; Carcinoma, Hepatocellular ; genetics ; Cocarcinogenesis ; Gene Expression Regulation, Neoplastic ; Genetic Variation ; Hepatitis B ; virology ; Hepatitis B virus ; Liver Neoplasms, Experimental ; genetics ; Point Mutation ; RNA, Neoplasm ; analysis ; Tumor Suppressor Protein p53 ; genetics ; Tupaiidae

OBJECTIVEThis study was designed to establish the two-dimensional electrophoresis profiles with high resolution and reproducibility from human lung squamous cell carcinoma tissue and paired tumor-adjacent normal bronchial epithelial tissue, and to identify differential expression of tumor-associated proteins by using proteome analysis.

METHODSComparative proteome analysis of human lung squamous carcinoma and paired normal bronchial mucosa adjacent to tumors from 20 cases were carried out. Total proteins of the carcinoma tissue and normal bronchial mucosa were separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were analyzed and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).

RESULTS(1) Seventy-six differentially expressed proteins were screened by analyzing the electrophoretic maps of the 20 carcinoma and control mucosa tissues. (2) Sixty-eight differential proteins were identified by peptide mass fingerprinting (PMF). Some proteins were products of oncogenes and others were involved in the regulation of cell cycle and signal transduction. (3) The expression of three proteins mdm2, c-Jun and EGFR, correlated with lung squamous carcinoma, were detected by immunohistochemical staining and Western blot analysis. The results showed that the expression of mdm2, c-Jun and EGFR were up-regulated in lung squamous carcinomas, whereas down-regulated in control normal mucosa. It was consistent with our proteome analysis results. Those results suggested that those proteins may play roles in the carcinogenesis of lung squamous carcinoma.

CONCLUSIONsixty-eight differentially expressed proteins were successfully characterized by comparative proteome analysis. Those results may provide scientific foundation for screening the molecular biomarkers which can be used in diagnosis and treatment of lung squamous carcinoma, as well as to improve patients' prognosis and provide a new clue for carcinogenesis research of lung squamous cell carcinoma.

Adult ; Aged ; Biomarkers, Tumor ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Proteome ; Proto-Oncogene Proteins c-jun ; metabolism ; Proto-Oncogene Proteins c-mdm2 ; metabolism ; Receptor, Epidermal Growth Factor ; metabolism ; Respiratory Mucosa ; metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Up-Regulation

Animals ; Cellular Reprogramming ; Dentate Gyrus ; cytology ; Fibroblasts ; cytology ; Mice ; Neural Stem Cells ; cytology ; transplantation ; Swine