Objective To investigate whether hepatocyte growth factor ( HGF ) and insulin-like growth factor (IGF1) induce cardiac stem cells (CSCs) to proliferate and directly differentiate into cardiomyocytes in vitro.Methods The myocardial tissues were dissected for primary culture of CSCs with the method of explants .The expressions of c-kit and CD34 were examined with immunofluorescence .Primary cells were purified with c-kit by flow cytometry.CFDA SE fluorescent probe was used to detect the proliferation of c-kit+CSCs.C-kit +CSCs were divided into two groups , and cardiac stem cells group and co-cultured with cardiomyocytes group , both group were cultured with HGF and IGF 1.An inverted microscope was used to observe changes in cell number and morphology in different periods .Living cells workstation was used to observe CFDA SE fluorescence intensity , to acquire images and do statistical analysis .Immunofluorescence technique was used to detect the expression of Nkx 2.5 and cardiac troponin T .Results In cardiac stem cells group ,CSCs had no obvious changes in cell number .In co-cultured with cardiomyocytes group , CSCs proliferated and had changes in morphology .Nkx2.5 and cTnT were positively expressed . Several CSCs differentiated into beating cardiomyocytes . Conclusion In co-cultured with cardiomyocytes condition , HGF and IGF1 may promote CSCs to proliferate and differentiate into beating cardiomyocytes .

This article was aimed to study the protection effects of Pa-Bing Formula No. 2 (PBFN-2) on neurons of substantia nigra in Parkinson's disease (PD) rats models in order to explore the possible mechanism of PBFN-2 in PD treatment. Rats were stereotaxically injected with 6-hydroxydopamine (6-OHDA) solution into the left stria-tum in two-site. Rat showed consistent right whirling and the number of rotation was more than 7 r·min-1 induced by APO in 30 min, then the rat was judged as PD model. A total of 14 rats modeled successfully were randomly di-vided into the model group and the treatment group. At the same time, the normal group and sham operation group were also established. Same volume of distilled water was given to the normal group, sham operation group and the model group. PBFN-2 (32.0 g·kg-1) was given to the treatment group for 4 weeks. Microscope was used to observe pathological changes of substantia nigra by nissl stain and changes of TH, Nrf2 and HO-1 immunohistochemical stain. The results showed that compared with the normal group, the nissl bodies were badly injured. Expressions of TH-positive cells were obviously reduced (P<0.01). The expression of Nrf2 nucleus protein and HO-1 protein were obviously increased in substantia nigra of PD rats in model group (P<0.01). Compared to the model group, PBFN-2 effectively increased nissl bodies in neuronal cells of substantia nigra of PD rats, and elevated the number of TH-immunoreactive cells in substantia nigra (P<0.05). The expressions of Nrf2 nucleus protein, HO-1 protein were significantly up-regulated (P<0.05). It was concluded that PBFN-2 had an obvious neuroprotection on the neuronal cells in substantia nigra of PD rats induced by 6-OHDA. The underlying mechanisms may be associated with regu-lation of Nrf2 nucleus protein and HO-1 protein expressions.

OBJECTIVE:To discuss the drug resistance situation and influential factors of sputum culture positive tuberculosis patients in a specialized subject hospital. METHODS:The information of sputum culture positive tuberculosis inpatients were col-lected from Heze infectious disease hospital during Jan. 1st 2014 to Apr. 30th 2015;the results of drug sensitive test were analyzed retrospectively by using absolute concentration method. RESULTS:Among 259 patients,137 cases were drug-resistant(52.90%), among including 69(26.64%) mono-drug resistant cases,52(20.08%) poly-drug resistant cases,15(5.79%)multi-drug resistant cases and 1(0.39%)extensively drug resistant case;streptomycin (31.66%) and amikacin (15.44%) were drugs with the highest drug-resistant rate;there was no statistical significance in resistant rate between different genders and ages(P>0.05),the rate of drug resistance in first-treatment patients was 47.21%,and that of the re-treatment patients was 70.97%,the difference was statisti-cal significant(P<0.05). In addition to ethambutol and kanamycin,other all had mono-drug resistance,there were 27 combinations of poly-drug resistance and 9 combinations of multi-drug resistance. Logistic regression analysis showed that treatment history was a risk factor for drug-resistance,the risk of re-treatment patient was 2.734 times of first-treatment patients. CONCLUSIONS:Sputum culture positive tuberculosis inpatients patients have high drug resistance and complex spectrum in the hospital;we should pay great attention to the risk of drug resistance in senile patients and re-treatment patients,and rational use of streptomycin,amikacin,isoni-azid and capreomycin.

Objective To study the relationship of three-dimensional power Doppler ultrasonographic parameters and endocrine profile in different symptoms of patients with polycystic ovary syndrome(PCOS).Methods One hundred and forty nine women with PCOS were divided into two groups,which included obese PCOS(OB-PCOS) group and non-obese PCOS (NOB-PCOS) group.The ultrasonic parameters such as follicle number,ovarian average diameter,ovarian volume,stromal volume,follicle volume,vascularization index(Ⅵ),flow index(FI),vascularization flow index(VFI) were measured and compared.Serum levels of luteinizing hormone (LH),follicle stimulating hormone (FSH),progesterone (P),estradiol (E2),testosterone(T),prolactin (PRL),sex hormone binding globulin (SHBG),free androgen index (FAI),fasting plasma glucose (FPG),fasting insulin (FINS),homeostasis model assessment-IR(HOMA-IR) were also measured and compared.The correlation of the ultrasonic parameters and hormonal factors were analyzed.Results The follicle number,ovarian average diameter,ovarian volume,stromal volume,follicle volume,FI and VFI,FINS,HOMA-IR,FAI of OB-PCOS were significantly higher than those of NOB-PCOS (P ＜0.01 or 0.05),the FSH,SHBG were significantly lower than those of NOB-PCOS (P ＜0.05 or 0.01).In OB-PCOS group,the follicle number was significantly associated with FSH(r =0.771,P ＜0.01).The ovarian volume,stromal volume,FI and VFI were significantly associated with HOMA-IR(r =0.412,0.842,0.389,0.415,P ＜0.05 or 0.01),FI was significantly associated with FAI (r =0.812,P ＜0.01).In NOB-PCOS group,the follicle number,ovarian volume were significantly associated with FAI(r =0.472,0.552,P ＜0.05)..Conclusions There are some different characters in ultrasonography and endocrine parameters between obese and non-obese PCOS patients.

Objective To explore the value of laparoscopy in the diagnosis and treatment of recurrent inguinal hernia in children.Methods The clinical data of 67 cases receiving laparoscopic treatment for recurrent inguinal hernia in children at the Department of General Surgery,Wuhan Children's Hospital,Tongji Medical College,Huazhong University of Science & Technology were retrospectively analyzed,including the type of hernia recurrence,operation time,intraoperative and postoperative complications,such as short-term hydrocele formation and testicular atrophy and so on.Results In 67 cases,there were 65 males and 2 females.Open repair surgery and laparoscopic surgery were respectively performed in 48 cases and 19 cases for the first operation and the hernia defects were found after operation.Contralateral patent vaginal process deformities were found in 11 sides in the reoperation.Among 67 cases of recurrent inguinal hernia,incomplete ligation of hernia sac was found in 21 cases,low level ligation of hernia sac in 17 cases,no ligation of hernia sac in 14 cases,omissive direct inguinal hernia in 4 cases,loose ligation of hernia sac in 4 cases,overlarge inner ring in 4 cases,weak abdominal muscles in 2 cases,and increased abdominal pressure (long-term constipation,asthma) in 1 case.All the patients were treated by laparoscopic hernia repair and no serious complications occurred during the operation.The average time of unilateral operation was (15.0±2.2) minutes (13-25 minutes),and bilateral operation was (27.0±4.3) minutes (18-41 minutes).All patients were discharged on the second day.Two patients showed short-term postoperative hydrocele and were cured with conservative treatment.During the follow-up time of (23±2) months (9-39 months),no recurrence or testicular atrophy was found.Conclusions Laparoscopy can confirm the type of recurrent inguinal hernia and contribute to reduce postoperative recurrence.Laparoscopic management of recurrent inguinal hernia in children is safe and feasible,which is expected to replace open hernia repair.

Objective To observe the effects of neuromuscular activation (Neurac) training on balance and walking in children with hemiplegic cerebral palsy. Methods 30 children with hemiplegic cerebral palsy from March to October, 2015 were divided into observation group (n=15) and control group (n=15). Both groups received routine rehabilitation, while the observation group received 30-minute Neurac training in addition, 5 times a week, for 3 months. They were assessed with D and E domains of Gross Motor Function Measure 88 (GM-FM-88), balance of Fugl-Meyer Assessment and footprints analysis before and after treatment. Results The scores of D and E domains of GMFM-88 and balance of Fugl-Meyer Assessment, and the step length, step width and velocity improved in both groups after training (t>7.31, P<0.001), especially in the observation group (t>2.08, P<0.05). Conclusion Neuroac training can further promote the recovery of gross motor function, balance and walking in children with hemiplegic cerebral palsy.

Aim To construct and express the eukary-otic expression vector of double-stranded RNA-depend-ent protein kinase (PKR)fusion green fluorescent and analyse its antiviral activity of HBV in vitro.Methods The PKR gene was cloned into an empty expression vector pEGFP-N1 using molecular clone technology. After being confirmed by restriction enzyme digestion and sequencing methods,the recombinant plasmid was named as pEGFP-PKR that was subsequently transfect-ed into HepG2.2.15 cells using LipofectamineTM2000. The expression level of PKR in HepG2.2.15 cells was confirmed by using fluorescent microscopy. Mean-while,HBV DNA and HBsAg/HBeAg were detected by real-time PCR and electrochemiluminescence meth-od,respectively.Results Both restriction enyme di-gestion and sequencing assays showed that the recombi-nant vector pEGFP-PKR was successfully constructed in our study.Fluorescent microscopy observation indi-cated that the fusion protein pEGFP-PKR expressed ef-ficiently in HepG2.2.15 cells.Moreover,compared with the empty vector group,the expression of HBV antigen in supernatants was significantly decreased (P<0.05 ).However,the extracellular HBV DNA ex-pression was not inhibited significantly.Conclusion In vitro,PKR proteion has certain antiviral activity of HBV.

Objective To investigate the analgesic effects of intraperitoneal lithium chloride injection on radicular pain behaviors in rats.Methods Using rat model of radicular pain induced by chronic compression of dorsal root ganglion(CCD) ,40 male SD rats were randomly divided into model group and Sham-operation group (group S, n= 12) of radicular pain were established.The rats in the model group were subdivided randomly into Control group(group C, n= 12) ,Early treatment group(group E, n=8) and Later treatment group(group L, n= 8 ).Rats in group E were intraperitoneal injected with lithium chloride once daily on day 2 ～ 4 after CCD respectively,while rats in L,group C and S treated with Vehicle(0.9% NaCl).Rats in L group were intraperitoneal treated with lithium chloride on day 12 ～ 14 after CCD respectively,while rats in E,group C and S received Vehicle.The pain ethology indexes such as paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were tested on day 1 before operation and day 1,4,7,10, 14, 17 and 21 after operation.Results Compared to S group and preoperative level, PWMT and PWTL decreased at Day 1 postoperative in group C (P＜0.05).At day 4 after the operation,compared with group C(7.712 ±0.237)g and (8.190 ±0.382) s,PWMT and PWTL of E group increased to ( 14.607 ± 0,280) g and ( 19.940 ± 0.933 ) s (P ＜ 0.05 ) after intraperitoneal injected lithium chloride.At day 14, compared with group C ( 6.788 ± 0.331 ) g and ( 7.301 ± 0.481 ) s, PWMT and PWTL of group L increased to ( 11.700 ± 0.379) 8 and ( 18.524 ± 1.060) s (P ＜ 0.05 ).This analgesic effect of lithium chloride continued to exist at day 21.However, there was still a significant difference between S group and E,group L(P＜0.05).Conclusion Intraperitoneal lithium chloride injection alleviates pain behavior on radicular pain in rats.

Objective To investigate the effects of intrathecally coadministered dexamethasone and spironolactone in trathecally on radicular pain behaviors.Methods Using rat model of radicular pain induced by chronic compression of dorsal root ganglion (CCD) ,48 male SD rats successfully received intrathecal catheter implantation and without motor dysfunction were randomly divided into Sham-operation group (Sham group, n= 12),Control group ( C group, n = 12 ), Dexamethasone group ( D group, n = 8 ), Spironolactone group ( S group, n = 8 )and Dexamethasone plus spironolactone group (DS group, n=8).Rats in D group,S group or DS group were intrathecally treated with dexamethasone 4 μg, spironolactone 3 μg or dexamethasone 4 μg plus spironolactone 3 μg twice daily on day 2 ～4 after CCD respectively,while rats in C and Sham group received 10μl 10% alcohol.Paw withdrawal mechanical threshold(PWMT) and paw withdrawal thermal latency (PWTL) were tested on day 1 before CCD and day 1,4,7,10,14,17 and 21 after CCD.Results Compared with Sham group, both PWMT and PWTL were significantly decreased after CCD surgery on the ipsilateral side(P＜0.01 ＝.Intrathecally administrated with dexamethasone significantly improved pain behaviors (P＜0.01 ＝ and these therapeutic effects lasted up to 10 days after CCD surgery.As with dexamethasone,intrathecal spironolactone also significantly attenuated PWMT (P＜0.01 ＝ and PWTL (P＜0.01 ＝ and the change lasted up to 7 days after CCD surgery.Coadministration spironolactone and dexamethasone exhibited significant synergies( PWMT: ( 13.52 ± 0.72) g vs ( 11.58 ± 1.38 ) g, P ＜0.01; PWTL: ( 19.63 ± 1.68) s vs ( 14.14 ± 1.52) s, P ＜ 0.01 ＝.These effects lasted up to at least 10 days.Conclusion Both dexamethasone and spironolactone intrathecally have therapeutic effects on radicular pain behaviors, combination injection of these two drugs could generate significant synergies.