OBJECTIVETo study the dynamic changes in the protein marker expression in the spermatogonial stem cells (SSCs) of mice at different ages by iTRAQ protein mass spectrometry and to screen new markers using the bioinformatic proteome database.

METHODSBased on the postnatal weeks, we divided 80 healthy male C57BL/6 mice into eight age groups of equal number, harvested their testicular tissues, extracted proteins following purification of the SSCs by compound enzyme digestion and magnetic-activated cell sorting. Then we analyzed and identified proteins using two-dimensional electrophoresis, protein mass spectrometry, and protein database information.

RESULTSTotally, 248,510 mass spectra were obtained from the MS experiment and 1132 proteins were identified. By the criteria of >1.2-fold for protein abundance difference and P value <0.05, we identified 298 differentially expressed proteins and 9 currently known makers of SSCs (PCNA, GFRalpha1, CDH1, Annexin A7, UCHL1, VASA, CD49f, CD29, and PLZf). Compara- tive analysis showed different expressions of the proteins in the SSCs of the mice of different ages, and the differences in the expressions of GFRalpha1, CD49f, and CD29 were consistent with the findings in other published literature. Ten proteins (P63, CD71, CD98, K19, ACE, K18, K15, K17, SH2, and SH3) were selected as SSC markers to be further studied.

CONCLUSIONThe proteins in SSCs are differentially expressed in mice of different ages. The technology of iTRAQ protein mass spectrometry can be used to analyze and compare the proteome information of mouse SSCs, obtain differentially expressed proteins in mice of different ages, and thus offers a new ap- proach to further analysis and study of the function and roles of these differential proteins.

Adult Stem Cells ; cytology ; metabolism ; Age Factors ; Animals ; Biomarkers ; analysis ; metabolism ; Cell Separation ; methods ; Electrophoresis, Gel, Two-Dimensional ; Male ; Mass Spectrometry ; Mice ; Mice, Inbred C57BL ; Proteins ; analysis ; metabolism ; Spermatogonia ; cytology

OBJECTIVETo evaluate the effect of autologous bone marrow stem cell transplantation in the treatment of severe liver damage.

METHODSAutologous bone marrow (50 ml) was harvested from 6 patients aged 44 to 69 years admitted for severe liver damage. Human bone marrow stem cells (HMSCs) were isolated and transplanted in to the patients' liver. At l, 4, 8, and 12 weeks after the transplantation, the changes in ALT, ALB, Cr, TB, PT and the clinical symptoms of the patients were observed.

RESULTSThe transplantation of autologous bone marrow stem cells resulted in obvious improvement of the liver function. At 12 weeks after the transplantation, ALT was reduced from 98.4 IU/L to 41.5 IU/L, TB from 136.5 µmol/L to 78.4 µmol/L, Cr from 112.3 µmol/L to 72.1 µmol/L, and ALB rose from 23.3 g/L to 32.6 g/L. The survival of the patients was 100% at 12 weeks, but one patient died at 7 months after the transplantation. The symptoms of the patients were also alleviated after the transplantation. At 12 weeks after transplantation, 3 patients reported improved appetite, 3 showed recovery of physical strength, and 2 showed lessened abdominal swelling. No serious adverse complications in association with the transplantation were found in the in 4 patients available to the follow-up.

CONCLUSIONAutologous bone marrow stem cell transplantation can improve the liver function of patients with severe liver damage without causing serious complications.

Adult ; Aged ; Bone Marrow Transplantation ; Hepatic Insufficiency ; surgery ; Humans ; Middle Aged ; Transplantation, Autologous ; Treatment Outcome

Objective To evaluate the effect of motivational interviewing(MI) on the rehabilitative exercise for patients after percutaneous coronary intervention (PCI).Methods 132 patients after percutaneous coronary intervention were chosen by the method of convenience sampling from a hospital in Suzhou,and were randomly divided into two groups.The experimental group (n =68) received MI based intervention while the control group (n =64) received routine health education.Results One month after discharge,there were 0,1,0,56 and 11 cases of patients in the experimental group and 0,10,7,35 and 12 cases in the control group who were respectively in the pre-contemplation stage,contemplation stage,preparation stage,action stage and maintenance stage of physical exercise,and the difference was statistically significant (Z =-2.23,P ＜ 0.05).Three months after discharge,there were 0,1,0,56 and 11 cases of patients in the experimental group and 13,12,2,25 and 12 cases in the control group who were respectively in the pre-contemplation stage,contemplation stage,preparation stage,action stage and maintenance stage of physical exercise,and the difference was statistically significant (Z =-3.62,P ＜0.05).Six months after discharge,there were 0,0,0,3 and 65 cases of patients in the experimental group and 20,8,1,1 and 34 cases in the control group who were respectively in the pre-contemplation stage,contemplation stage,preparation stage,action stage and maintenance stage of physical exercise,and the difference was statistically significant (Z =-5.70,P ＜ 0.05).Patients' exercising frequency were all no less than 5 times per week one month,three months and six months after discharge in the experimental group,and were significantly different from those in the control group (x2 =-4.22,-6.10,-7.12,respectively;P ＜0.01).There was also significant difference of duration of physical exercises between two groups (x2 =-4.96,-5.93,-7.24,respectively; P ＜ 0.01).Conclusions Motivational interview can effectively promote patients' willingness to enhance physical exercise and improve their exercise compliance after percutaneous coronary intervention.

OBJECTIVETo investigate the ultrasonographic features of obstructive azoospermia and to evaluate transrectal ultrasonography in the diagnosis of the disease.

METHODSWe performed transrectal ultrasonography for 248 patients with obstructive azoospermia, observed the sonographic features of the prostate gland, seminal vesicle and ejaculatory duct.

RESULTSThe average volume of the prostate gland of the studied group was 13.2 ml. A total of 111 cases showed normal sonographic features, 39 cases bilateral seminal vesicle defect, 33 cases bilateral seminal vesicle aplasia, 23 cases unilateral seminal vesicle defect and contralateral seminal vesicle aplasia, 28 cases bilateral and 14 cases unilateral seminal vesicle dilatation. Of the 42 cases of seminal vesicle dilatation, 18 had ejaculatory duct dilatation and 17 had cysts in the midline of the prostate.

CONCLUSIONObstructive azoospermia varies in kind. Transrectal ultrasonography can reveal the details of the prostate, seminal vesicle and ejaculatory duct and help to classify obstructive azoospermia and determine the location of the lesion.

Adult ; Azoospermia ; diagnostic imaging ; Ejaculatory Ducts ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Posture ; Prostate ; diagnostic imaging ; Rectum ; Seminal Vesicles ; diagnostic imaging ; Ultrasonography

OBJECTIVETo investigate the effects of single heat stress treatment on spermatogenic cells in mice.

METHODSWe randomly divided 36 C57 male mice into a control and a heat stress treatment group and submerged the lower part of the torso in water at 25 °C and 43 °C, respectively, both for 15 minutes. At 1, 7, and 14 days after treatment, we obtained the testicular organ indexes, observed the changes in testicular morphology by HE staining, and determined the location and expression levels of the promyelocytic leukemia zinc finger (PLZF) and synaptonemal comlex protein-3 (SCP-3) in the testis tissue by immunohistochemistry and Western blot.

RESULTSThe testicular organ index was significantly lower in the heat stress treatment than in the control group (P < 0.05). Compared with the controls, the heat shock-treated mice showed loosely arranged spermatogenic cells scattered in the seminiferous tubules at 1 day after heat stress treatment, atrophied, loosely arranged and obviously reduced number of spermatogenic cells at 7 days, and relatively closely arranged seminiferous tubules and increased number and layers of spermatogenic cells at 14 days. The number of SCP-3 labelled spermatocytes obviously decreased in the heat stress-treated animals at 1 and 7 days and began to increase at 14 days. The PLZF protein expression was significantly reduced in the heat stress treatment group at 1 day as compared with that in the control (0.19 ± 0.12 vs 0.64 ± 0.03, P < 0.01), but elevated to 0.77 ± 0.02 at 7 and 14 days, even remarkably higher than in the control animals (P < 0.01).

CONCLUSIONHeat stress treatment can induce short-term dyszoospermia in mice, which can be recovered with the prolonged time after treatment.

Animals ; Blotting, Western ; Hot Temperature ; Immunohistochemistry ; Male ; Mice ; Nuclear Proteins ; metabolism ; Promyelocytic Leukemia Protein ; Seminiferous Tubules ; cytology ; Spermatocytes ; cytology ; pathology ; Testis ; metabolism ; Transcription Factors ; metabolism ; Tumor Suppressor Proteins ; metabolism

OBJECTIVETo investigate the central mechanism of hypoglycemic effect induced by ear acupuncture.

METHODSExtracellular single-unit recordings were carried out in nucleus tractus solitarius (NTS) of rats after infusion of glucose, insulin via jugular venous and electroacupuncture at auricular concha, observing responses of glucose-sensitive neurons and insulin-sensitive neurons in NTS to electroacupuncture stimulation at auricular concha.

RESULTSThere existed glucose-sensitive neurons and insulin-sensitive neurons in NTS. Among glucose-sensitive neurons, inhibitory ones accounted for 37.3%, and excitable ones accounted for 10.9%. Among the insulin-sensitive neurons, excitable ones accounted for 33.3%, and inhibitory ones accounted for 4.9%. When electroacupuncture was given at auricular concha, 49.3% (34/69) of total neurons recorded in NTS showed increase in firing rate, while 4.3% (3/69) showed decrease in firing rate. Among the 34 exciting-responsive neurons, there were 18 neurons responding to glucose infusion with a discharge decreased in frequency, and there were 8 neurons responding to insulin infusion with a discharge increased in firing rate.

CONCLUSIONAcupuncture stimulation at auricular concha can irritate the neurons in NTS, which show largely inhibitory and excitable response to glucose and insulin infusion, respectively. The hypoglycemic effect induced by auricular acupuncture possibly results from regulating these neurons' firing activities.

Acupuncture, Ear ; Animals ; Electroacupuncture ; Glucose ; pharmacology ; Insulin ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley ; Solitary Nucleus ; drug effects ; physiology

To investigate the relationship between HLA-DQB1* alleles and major beta-thalassemia, the HLA-DQB1* loci typing was performed with polymerase chain reaction-sequence specific primer (PCR-SSP) in 42 unrelated (unconsanguineous) patients with major beta-thalassemia and 45 normal control individuals in Guangdong Province. Results showed that the frequency of HLA-DQB1*06 allele in patient group (19.0%) was higher than that in the control group (4.4%) kappa(2) = 8.961, p < 0.01). Our data suggests that HLA-DQB1*06 allele is associated with pathogenesis of the major beta-thalassemia in Guangdong area.
Alleles ; Asian Continental Ancestry Group ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; HLA-DQ Antigens ; genetics ; HLA-DQ beta-Chains ; Humans ; Male ; beta-Thalassemia ; genetics

OBJECTIVETo explore the clinical efficacy of the percutaneous kyphoplasty for the treatment of osteoporotic Kummell's disease.

METHODSFrom May 2010 to February 2012, 8 patients with osteoporotic Kummell's disease were treated with percutaneous balloon kyphoplasty. There were 2 males and 6 females, with a mean age of 73.4 years. All the patients suffered from lower back pain for 4.7 months, which affected seriously the patient's quality of life. The anterior vertebral height and Cobb angel was measured on standing lateral radiograph at pre- and post-operatively (2 days after operation) and during the final follow-up. Visual analog scale (VAS), Japanese Orthopedic Association (JOA) and Oswestry disability index (ODI) were used to evaluate pain and function.

RESULTSIncision of all patients healed normally without infection. The level of back pain decreased remarkably after surgery. The mean time of ambulation was 4.3 days (ranged, 1 to 10 days). Cement leakage occurred in 1 case with no symptom. The anterior vertebral height and Cobb angel of the fractured vertebra recovered respectively from preoperative (30.4 +/- 7.4)% and (31.3 +/- 9.9) degree to (70.3 +/- 3.3)% and (9.1 +/- 3.0) degree at the 2nd day after operation. VAS and JOA scores, ODI improved from preoperative 8.7 +/- 1.2, 12.3 +/- 1.7 and (93.3 +/- 4.6)% to 3.1 +/- 1.1, 24.9 +/- 1.6 and (32.2 +/- 5.4)% respectively at the 2nd day after operation. All the patients were followed up, and the duration ranged from 3 to 24 months, with a mean of 4.7 months. At the latest follow-up, the anterior vertebral height and Cobb angel of the fractured vertebra were (69.9 +/- 3.2)% and (10.9 +/- 2.4) degree; the VAS and JOA scores and ODI were 2.2 +/- 1.0, 26.4 +/- 1.4 and (29.2 +/- 4.5)% respectively, which had no obvious difference compared to those results at the 2nd day after operation.

CONCLUSIONBalloon kyphoplasty is safe and effective treatment for osteoporotic Kummell's disease.

Aged ; Aged, 80 and over ; Female ; Humans ; Kyphoplasty ; methods ; Male ; Osteoporosis ; surgery ; Treatment Outcome

Objective To assess the effect of silent mating type information regulation 2 homolog 6 (SIRT6) on migratory and proliferative activity of skin fibroblasts in the elderly,and to explore their mechanisms.Methods Circumcised foreskins were obtained from patients of different ages in Department of Urologic Surgery,Second Hospital of Shanxi Medical University,including 8 elderly patients and 8 young patients.Human skin fibroblasts were isolated from the foreskin tissues by using a collagenase digestion method.Western blot analysis was performed to determine the expression of SIRT6 and phosphorylated p65 (p-p65) in human skin fibroblasts in different age groups,scratch assay to evaluate cell migratory activity,and cell counting kit-8 (CCK8) assay to assess cellular proliferative activity.Skin fibroblasts in the elderly group were divided into 2 groups:SIRT6 group transfected with a lentiviral vector overexpressing SIRT6,and control group transfected with an empty lentiviral vector.Then,the cell migratory and proliferative activity as well as p-p65 expression were measured by the above methods,and the mRNA expression of type Ⅰ and Ⅲ collagens,and integrin subunits α3,α5 and β1 was determined by real-time PCR in the SIRT6 group and control group.Statistical analysis was carried out with GraphPad Prism 5 software by using t test for comparison between 2 groups.Results Compared with the young group,the elderly group showed significantly decreased SIRT6 expression in skin fibroblasts (0.434 ±0.179 vs.1.000 ± 0.067,t =3.040,P =0.012),migration rate (43.81％ ± 18.84％ vs.94.63％ ± 12.32％,t =5.903,P =0.003)and cellular proliferative activity at 24 and 48 hours (both P ＜ 0.05),but significantly increased p-p65 expression (1.694 ± 0.148 vs.1.000 ± 0.093,t =2.949,P =0.015).Compared with the control group,the SIRT6 group showed significantly decreased p-p65 expression (P ＜ 0.05),but significantly increased migratory and proliferative activity (both P ＜ 0.05),and elevated mRNA expression of type Ⅲ collagen and integrin subunits oα3,α5 and β1 (all P ＜ 0.05).Conclusion SIRT6 can improve the migratory and proliferative activity of human fibroblasts in the elderly,possibly by inhibiting the nuclear factor-κB pathway.

Objective To develop a questionnaire for measuring prevention motivation for nursing staffs' needlestick injuries based on the Planned Behavior Theory and evaluate its reliability and validity.Methods The questionnaire development of prevention motivation for nursing staffs' needlestick injuries was based on the framework of Planned Behavior Theory.Questionnaire items were constructed by literature reviewing,and screened by Delphi Method and pilot test.Totals of 320 frontline clinical nurses in Suzhou were selected to evaluate the reliability and validity of the questionnaire.Results Totals of 306 questionnaires were recovered,with the rate of 95.63％.Totals of 300 questionnaires were qualified,with the rate of 98.04％.The questionnaire of prevention motvation for nursing staffs' needlestick injuries was composed of four dimensions:preventive behavior belief,result evaluation,subjective norm and perceived behavior control.Cronbach' sα coefficients of the questionnaire were 0.83,test-retest reliability was 0.94 and content validity index (CVI) was 0.91.The correlation coefficient r between every dimension and the questionnaire was 0.484-0.705(P ＜0.01),the correlation coefficientr between dimensions was 0.339-0.680 (P ＜ 0.01),the cumulative contribution of variance was 60.743％,and the questionnaire had favorable reliability and validity.Conclusions The questionnaire is proved to have excellent level of reliability and validity,which can effectively explain and predict the level of prevention motivation for nursing staffs' needlestick injuries,as well as provide nursing leaders with references for targeting prevention education intervention.