Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Myocytes, Cardiac ; cytology

OBJECTIVETo assess the effect of Klotho gene transduction on the progression of hypertension and heart damage in spontaneous hypertensive rats (SHRs).

METHODSAn adeno-associated virus (AAV) carrying full-length mouse Klotho cDNA (rAAV.mKL) was constructed for in vivo expression of Klotho. Three different groups of male SHRs and a control group of sex and age-matched Sprague Dawley (SD) rats (5 rats per group) were used. The experimental groups of SHRs received an IV injection of phosphate buffered saline (PBS), rAAV.mKL and rAAV.EGFP, respectively. The control group only received equal-volume of PBS. The whole study has spanned 12 weeks. Plasma levels of insulin-like growth factor-1 (IGF-1) and insulin were measured with ELISA. The weight of whole heart was measured to calculate the heart weight index (HWI). EGFP expression of heart frozen sections was observed by fluorescence microscopy. Expression of mRNA and protein of Klotho, IGF-1, IGF-1 receptor (IGF-1R) and p-Akt were determined with RT-PCR, immunohistochemical analysis, and Western blotting. Hypertrophic myocardial cell and collagen fiber were observed by histological examination following Haematoxylin-Eosin and Masson staining.

RESULTSTransduction of rAAV.mKL can significantly prevent the increase of blood pressure in SHRs. Compared with the control group, the levels of Klotho mRNA and protein have both increased, and the plasma levels of IGF-1, insulin and glucose were elevated, whereas the expression of phosphor-Akt (also called Protein Kinase B, PKB) was decreased in the rAAV.mKL group. Furthermore, a decrease of hypertrophic myocardial cells and collagen fibers was noticed in the rAAV.mKL group compared with the control group.

CONCLUSIONThe Klotho gene can attenuate the progression of hypertension and abolishes myocardial hypertrophy and myocardial fibrosis. The protective effect observed in the rAAV.mKL group of SHRs may be attributed to increased Klotho protein and suppression of insulin and IGF-1 signaling pathways through inhibition of Akt phosphorylation.

Animals ; Blood Glucose ; Blood Pressure ; genetics ; Gene Expression ; Glucuronidase ; genetics ; metabolism ; Hypertension ; genetics ; metabolism ; pathology ; Insulin ; blood ; Insulin-Like Growth Factor I ; genetics ; metabolism ; Male ; Myocardium ; metabolism ; pathology ; ultrastructure ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Inbred SHR ; Transduction, Genetic

Reduced glutathione (GSH) , the most abundant non-protein thiol in mammalian cells, exists extensively in the human body and plays a key role in many biological processes, including the synthesis of proteins and DNA and the transport of amino acids, especially in protecting cells against oxidation. GSH also shows its important clinical value in the treatment of many diseases. It has been reported that GSH also widely exists in the male reproductive system and has a therapeutic role in male infertility. The objective of this review is to summarize the distribution of GSH in the male reproductive system and its therapeutic value for male infertility.
Genitalia, Male ; metabolism ; Glutathione ; metabolism ; therapeutic use ; Humans ; Infertility, Male ; therapy ; Male ; Reproductive Techniques, Assisted

Aim To explore the effect of Klotho (KL) gene transfection on the apoptosis of MC3T3-E1 osteo-blasts induced by dexamethasone(DEX). Methods MC3T3-E1 osteoblasts were transfected by recombinant adenovirus containing KL gene(Ad-KL) and recombi-nant adenovirus containing green fluorescent protein (GFP) gene(Ad-GFP). The apoptosis model was con-structed. The transfection efficiency of Ad-KL and Ad-GFP in cells were observed using inverted fluorescent microscope, and the level of KL mRNA and protein was detected by qPCR and Western blot,respectively. The cell viability after different concentrations of DEX acting on the cells and the viability of every research group were determined by cell counting kit-8 (CCK-8) assay. The apoptotic rate was evaluated by flow cytom-etry. The level of mRNA and protein was analyzed by qPCR and Western blot, respectively. The level of caspase-9 protein was detected by immunofluorens-cence assay. Results Cells were transfected by Ad-KL and Ad-GFP successfully. KL group and KL +DEX group had higher level of KL mRNA and protein than that in other groups. The optimum concentration of DEX was 2.0 mmol·L-1. When DEX acting on the cells, the cells viability decreased and apoptotic rate increased obviously in DEX group and GFP + DEX group. The level of Bax mRNA and protein presented a upward trend in DEX group and GFP +DEX group, while the level of Bcl-2 mRNA and protein was oppo-site. But after KL transfecting MC3T3-E1 osteoblasts, the markers described above in KL group had more dramatic improvement than in DEX group and KL +DEX group. Conclusions High-dosage DEX can in-duce the apoptosis of MC3T3-E1 osteoblasts, and the pro-apoptosis effect of high-dosage DEX in MC3T3-E1 osteoblasts can be suppressed by up-regulating KL gene expression level, suggesting that the glucocorticoid-in-duced osteoporosis might be improved by up-regulating KL gene expression level, and it may be a new target for the treatment of latrogenic osteoporosis induced by high-dosage glucocorticoid in clinic.

OBJECTIVETo discuss the feasibility of the operation of minimal invasive with gallbladder preserved via choledochoscopy.

METHODSFrom February 1992 to June 2006, there were 760 patients who underwent cholecystolithiasis treated with the minimal invasive operation with gallbladder preserved via choledochoscopy, among which there were 428 males and 332 females, aged from 18 to 81 years old. All cases were diagnosed by ultrasonography and their gallbladder functions were proved normal by the examination of oral cholecystography or ECT before operation. In the operation gallstones were removed from gallbladder completely.

RESULTSThere were 612 cases who were followed up for 1-15 years and the follow-up rate was 80.5%. All patients recovered well after operation. The post-operation rate of recurrence of gallstone was 0.49%, 4.39%, 5.83%, 6.60%, 7.21% and 8.38% within the first year, the second year, the third year, the fifth year, the seventh year and the ninth year respectively, rate of recurrence of gallstone were 10.11% within both the tenth and the fifteenth year.

CONCLUSIONSThe minimal invasive operation with gallbladder preserved via choledochoscopy is effective to cholecystolithiasis patients whose gallbladder function is normal. It is a feasible operation that preserves the normal functional gallbladder and improves the patients' life quality.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cholecystolithiasis ; surgery ; Endoscopy, Digestive System ; methods ; Feasibility Studies ; Female ; Follow-Up Studies ; Gallbladder ; surgery ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo analyze the clinical characteristics of 267 cases with occupational chronic carbon disulfide (CS(2)) poisoning and to provide the basis for revising the items of periodical medical examination of workers occupationally exposed to CS(2).

METHODSThe subjects of present study were 267 patients with mild CS(2) poisoning diagnosed according to "Diagnostic Criteria of Occupational Chronic Carbon Disulfide Poisoning (GBZ4-2002)" from April in 2006 to May in 2010. All patients were from the same chemical fiber factory. When a subject was diagnosed as patient with CS(2) poisoning, who should interview with questionnaire which included the illness and occupational history, symptoms, individual habits. The physical examination, nervous test, cardiovascular test, biochemical test and electromyogram were performed.

RESULTSThe rate of decreased motor conduction velocity was 87.3% (233/267 roots). The highest detection rate of slowing conduction velocity was the common peroneal motor nerve which was 48.6% (138/248 roots) and the second was median motor nerve with delay rate of 37% (155/419 roots). The main symptoms of the patients were neurasthenia, numbness and paresthesia. The rates of abnormal achilles tendon reflex and knee jerk reflex in patients were were 79.4% and 49.8%, respectively. The detected rates of patients with ST-segment changes and hypertension were 19.1% and 27.5%, respectively. The rates of hypertension, systolic pressure and diastolic pressure were 27.3%, 22.5% and 21.1%, respectively. The rates of lactate dehydrogenase (LDH), triglycerides (TG) and low density lipoprotein (LDL) were high. The detected rates of urine acid, indirect bilirubin and total bilirubin in male patients were higher than those in female patients. In addition, the abnormal detected rate of urea nitrogen and indirect bilirubin increased with exposure years.

CONCLUSIONOccupational chronic CS(2) poisoning mainly affects the nervous system, as well as liver and kidney function. Detecting the median and common peroneal motor nerve conduction velocities could be the screening indicators for the peripheral nerve injury induced by CS(2) in the occupational exposure population during the periodical occupational medical examinations.

Adult ; Aged ; Carbon Disulfide ; poisoning ; Chemical Industry ; Female ; Humans ; Kidney ; drug effects ; physiopathology ; Liver ; drug effects ; physiopathology ; Male ; Middle Aged ; Multiphasic Screening ; Nervous System ; drug effects ; physiopathology ; Neural Conduction ; Occupational Exposure

OBJECTIVETo investigate the feasibility and clinical significance of detecting sperm mitochondrial membrane potential (MMP) by JC-1 fluorescent staining and flow cytometry, and to explore the relationship between the results of JC-1 staining and seminal parameters.

METHODSSixty-three semen samples were divided into a fertile (n = 31) and an infertile group (n = 32) and underwent computer-assisted semen analysis (CASA). All the samples were washed, followed by JC-1 staining and evaluation of sperm MMP by flow cytometry. The percentage of normal sperm MMP was indicated as the percentage of sperm emitting orange-red fluorescence (JC-1 + %).

RESULTSThe JC-1 + % was significantly higher in the fertile group than in the infertile one ([75.89 +/- 15.69]% vs [54.04 +/- 22.21] %, P = 0.000), correlated positively with sperm motility (r = 0.610, P = 0.000) and the percentage of grade a + b sperm (r = 0.614, P = 0.000) and negatively with grade d sperm (r = -0.504, P = 0.000). There was a significant positive correlation between the results of JC-1 staining (JC-1 + %) and that of Rh123 /PI dual fluorescent staining (Rh123 + / PI (-)%) (r = 0.938, P = 0.000).

CONCLUSIONJC-1 staining and flow cytometry could readily and quickly detect sperm MMP and the sperm JC-1 + % could be an auxiliary marker for the diagnosis of male infertility.

Adult ; Flow Cytometry ; methods ; Fluorescent Dyes ; chemistry ; Humans ; Male ; Membrane Potential, Mitochondrial ; physiology ; Middle Aged ; Reproducibility of Results ; Sperm Motility ; physiology ; Spermatozoa ; physiology ; Staining and Labeling ; methods

BACKGROUNDChromosomal abnormalities have been shown to play an important prognostic role in multiple myeloma (MM). Interphase fluorescence in situ hybridization (i-FISH) has been much more effective to identify cytogenetic aberrations in MM than conventional cytogenetic technique (CC). To clearly determine the cytogenetic features of Chinese MM patients and identify their prognostic implications, we designed a multicenter study based on i-FISH including 672 patients from 52 hospitals in China.

METHODSAll 672 patients were systematically screened for the following genomic aberrations: del(13q), IgH rearrangement, del(p53) and 1q21 amplifications.

RESULTSThe analysis showed that the chromosomal changes were detected in 22.1% patients by CC and in 82.3% patients by i-FISH. The most common abnormalities by CC were chromosome 1 aberrations (48.4%), -13/13q- (37.6%), hyperdiploidy (36.6%), hypodiploidy (30.1%) and IgH rearrangements (23.7%). The most frequent abnormalities by FISH was del(13q), which was found in 60.4% patients, whereas IgH rearrangement, 1q21 amplification and p53 deletions were detected in 57.6%, 49.0% and 34.7% cases, respectively. By statistical analysis, -13/13q- by CC was associated with low level of platelet (P = 0.015), hyperdiploidy was associated with low level of serum albumin (P = 0.028), and IgH rearrangement by FISH was associated with high level of β2 microglobulin (P = 0.019). Moreover, 1q21 amplification and del(p53) by FISH conferred a high incidence of progressive disease (PD) after initial therapy. Metaphase detection of IgH rearrangements and chromosome 1 aberrations concurrently was associated with a short progression free survival (PFS) (P = 0.036). No significant prognostic implications of other cytogenetic abnormalities were found associated with overall survival and PFS.

CONCLUSIONSChinese MM patients had similar cytogenetic abnormalities compared with the previous reported studies. However, the prognostic significance of FISH aberrations were not clearly determined and further study is required.

Adult ; China ; Chromosome Aberrations ; Chromosomes, Human, Pair 1 ; genetics ; Cytogenetic Analysis ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology

OBJECTIVETo analyze the clinical, molecular and cytogenetic features of 46, XX (SRY positive) male syndrome.

METHODSThe clinical features of 4 patients with 46, XX (SRY positive) male syndrome were analyzed retrospectively. Karyotyping, FISH, PCR amplification of the SRY gene, and Y-chromosome microdeletion were performed to study their molecular cytogenetic features.

RESULTSThe Four patients were all sociopsychologically males of short stature and came to hospital for infertility. Physical examination revealed that their testes were small in volume and soft in texture, but their penes were normal. Semen analyses showed complete azoospermia. Detection of serum sexual hormone suggested hypergonadotropic hypogonadism. All were karyotyped as 46, XX. Molecular analyses revealed the presence of the SRY gene and absence of AZFa, b and c of the Y chromosome. FISH analysis showed that SRY genes were translocated to Xp in 3 of the patients.

CONCLUSIONPhenotypically 46, XX (SRY positive) male patients are males generally, for the presence of the SRY gene in the whole genome and azoospermia due to the deletion of AZF. The clinical characteristics of the patient include testis dysgenesis, infertility and short stature. The long arm of the Y chromosome might contain the gene associated with body height. Extensive molecular and cytogenetic studies on 46, XX male syndrome may help to elucidate its genotype-phenotype relation.

Adult ; Body Height ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Estradiol ; blood ; Follicle Stimulating Hormone ; blood ; Genes, sry ; Gonadal Dysgenesis, 46,XX ; blood ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Luteinizing Hormone ; blood ; Male ; Polymerase Chain Reaction ; Prolactin ; blood ; Syndrome

OBJECTIVETo determine the level of uric acid (UA) in the expressed prostatic secretion (EPS) of chronic prostatitis patients and explore its clinical significance.

METHODSA total of 91 patients with chronic prostatitis diagnosed by NIH standard were divided into a III A (n = 48) and a III B (n = 43) group, and healthy volunteers were selected as the control. The scores on the NIH-Chronic Prostatitis Symptom Index (CPSI) and the WBC count, pH value and UA level in EPS were evaluated for all the three groups.

RESULTSThe EPS UA concentration was (257.02 +/- 144.84) micromol/L in Group III B, significantly higher than in Group III A, (159. 73 +/- 121.49) micromol/L, (P < 0.01), and the control, (78.55 +/- 44.53) micromol/L, (P < 0.01). The level of EPS UA was correlated negatively with pH value (r = -0.398, P = 0.000), but positively with CPSI-P, CPSI-U and CPSI-T (r = 0.436, 0.316 and 0.403, P < 0.01).

CONCLUSIONBackflow of urine into prostatic ducts might cause chemical inflammation reaction by increasing UA concentration. There is a close relationship between the UA level in EPS and chronic prostatitis symptoms. Determination of the UA level in EPS is of great significance for the diagnosis and treatment of chronic prostatitis.

Adolescent ; Adult ; Chronic Disease ; Humans ; Male ; Middle Aged ; Prostate ; pathology ; secretion ; Prostatitis ; diagnosis ; metabolism ; physiopathology ; Uric Acid ; analysis