Objective To investigate the effects of matrine on proliferation and apoptosis of human rhabdomyosarcoma(RMS) RD cells line in vitro,to study the regulatory mechanism of Wnt/β-catenin pathway-influenced apoptosis of RD cells line by detecting the expressions of β-catenin protein,Bcl-2 protein and caspase-3 protein,and to explore Wnt/β-catenin mechanism during the process of RMS.Methods The human RMS RD cells line was treated with matrine of different concentrations (0.5,1.0,1.5,2.0 g/L)for 48 hours respectively,and the proliferation inhibition rates of different concentrations of matrine on RD cells were detected by methyl thiazolyl tetrazolium assay,while the apoptosis rates by flow cytometry (FCM) and the expressions of β-catenin,Bcl-2 and caspase-3 by Western blot.Results The proliferation inhibition rates between control group and different concentrations of matrine groups were(13.70 ±0.25)％,(33.16 ±0.11)％,(42.96 ±0.90)％,(56.26 ±0.79)％ and (67.89 ±0.63)％,respectively.The apoptosis rates were (5.49 ± 0.96) ％,(17.23 ± 5.03) ％,(25.84 ± 4.17) ％,(36.08 ± 3.68) ％and (47.79 ± 4.82) ％,respectively.The highest expression of β-catenin and Bcl-2 proteins and the minimum amount of caspase-3 protein were found in the control group.After intervention of matrine,the expressions of β-catenin and Bcl-2 reduced while the amount of caspase-3 rose significantly,which was concentration-dependent obviously.Differences were found between every concentration of matrine group with control group according to statistics (all P ＜ 0.05).Conclusions Matrine can inhibit proliferation and induce apoptosis of RD cells.Matrine can down-regulate the expression of β-catenin and Bcl-2 proteins in RD cells,while the amount of caspase-3 protein rises.Wnt/β-catenin signal pathway plays an important role in the apoptosis of RD cell induced by matrine,and its downstream proteins Bcl-2 and caspase-3 are also involved in the regulation of this process.

@@

BACKGROUND:Periprosthetic femoral fracture is a severe complication after total hip arthroplasty. More than 85% belongs to Vancouver B2 and B3 fractures, and the operation is difficult. OBJECTIVE:To explore the clinical effect of Cable-Ready fast system and biotype long-stem prosthesis and its effect on the recovery of joint function in patients with Vancouver B2 and B3 type periprosthetic femoral fractures after total hip arthroplasty. METHODS: A total of 60 patients receiving total hip arthroplasty suffered from Vancouver B2 and B3 type periprosthetic femoral fractures in the Huanggang Central Hospital from September 2011 to September 2012. They were equaly divided into control and observation groups according to different fixation methods. Patients in the control group were treated by ordinary steel wire cerclage fixation combined with uncemented long-stem prosthesis; those in the observation group were treated by Cable-Ready fast system combined with fast uncemented long-stem prosthesis. RESULTS AND CONCLUSION:The average operation time, the time of hospitalization and fracture healing time were shorter in the observation group than in the control group. Moreover, the intraoperative blood loss was less in the observation group than in the control group. After 1 year of folow-up, Harris hip score was higher in the observation group compared with pre-treatment (t=3.174 9,P=0.002 6), and significantly higher than the control group (t=2.479 8,P=0.015 4). The excelent and good rate of Harris hip score was significantly higher in the observation group than in the control group (χ2=11.294 5,P=0.002 6). The total incidence of complications was significantly lower in the observation group than in the control group (χ2=8.139 7,P=0.04 2). These data indicate that Cable-Ready fast system combined with uncemented long-stem prosthesis in the treatment of Vancouver B2 and B3 periprosthetic femoral fractures after total hip arthroplasty has smal injury, less postoperative complications and better recovery of hip function after operation.

Objective To clone the human Islet neogenesis associated protein(rhINGAP)gene,express the gene extraeellulary in Pichia yeast for.further study on biological function and animal test on INGAP.Methods INGAP gene Was amplified with PCR and inserted into the recombinant plasmidα/pUC18.Then,the fusion gene of α and INGAP was digested and inserted into the expression plasmid pPIC9K.The positive recombinant plasmid which integrated INGAP Was confirmed by restriction enzyme digestion and sequencing,and it Was linearized with Sal Ⅰ digestion and transfered into the yeast host strain GS115 through electroporation.The yeast transformants that harbor the desired gene INGAP with high copy were selected by the auxotroph mediam G418,and verified by PCR.The condition of hake-flask culture was optimized,and the recombinant human INGAP Was induced expression with methanol as the only Carbone source.The antigen activity of the desired protein Was detected by Western blotting and ELISA method.Results Recombinant plasmid αINGAP/pPIC9K were successfully constructed and three positive Pichia yeast transformants were obtained.The expressed protein had satisfactory antigen activity,which Was confirmed by the Western blotting and ELISA method.Conclusions Pichia yeast expressing human Islet neogenesis associated protein (rhINGAP)gene was successfully constructed.

Objective To investigate the efficacy and safety of ultra-mini percutaneous nephrolithotomy for the treatment of renal or upper ureteral stones.Methods The data of 32 patients diagnosed as renal or upper ureteral stones were retrospectively reviewed,who underwent ultra-mini percutaneous nephrolithotomy from June to October,2015,including 29 kidney stones,3 upper ureteral stones,of which 21 single stone,7 multiple stones,and 4 staghorn stones.There were 8 cases with inferior calyx stones,10 with renal pelvis stones,3 with upper ureteral stones,10 with renal pelvis stones plus inferior calyx stones and 1 with renal pelvis stones plus upper calyx stones.The mean stone size was (20.1 ±7.6) mm (ranging from 10 to 41mm).The mean Hounsfield unit (HU) was (1 125.9 ±225.9) Hu (ranging from 520 to 1 550Hu).In this cohort,13 cases had mild hydronephrosis,2 moderate hydronephrosis,1 severe hydronephrosis while no hydronephrosis identified in the other 16 patients.One case had concomitant kidney and ureter duplication malformation,and 1 case had residual stones after laparoscopic nephrolithotomy.Results All of the patients were treated by single tract UMP.Among them,middle calyceal puncture was performed in 9 cases,and the other 23 cases underwent lower calyceal puncture,including 4 cases of intercostal puncture,and 28 subcostal puncture.The mean operating time was (30.3 ± 15.0) min (ranging from 10 to 90 min).The mean postoperative hospital stay was (1.9 ± 1.0)days (ranging from 1 to 5 days).The mean hemoglobin decrease was(14.6 ±8.4)g/L (ranging from 1 to 46 g/L).No analgesics were used.Peri-operative complication rate was 3.13％ (1/32) with 1 case of collection system perforation.There was no fever,severe bleeding,urinary extravasation,pleural injury,or blood transfusion.The SFR was 93.75％ (30/32) and 96.88％ (31/32) on the first day and the first month after the operation,respectively.Conclusions UMP could be a safe and effective method for the treatment of renal or upper ureteral stones,especially for inferior calyceal stones less than 2cm.It offers high stone clearance rate,minimally invasiveness,quick recovery,short length of hospital stay and improved quality of life.

Objective To discuss the management of renal cell carcinoma(RCC) associated with von Hippel-Lindau(VHL) disease. Methods Clinical data were analyzed retrospectively from 28 ca-ses ( 16 males and 12 females, with a mean age of 45 years), of whom 15 had bilateral RCC and 13 had unilateral RCC. VHL germline mutation was analyzed in 25 cases. Nephron sparing surgery (NSS) or radical nephrectomy was performed in 24 cases. Results VHL germline mutations were detected in 25 cases including 14 asymptomatic patients. Among 29 solid renal tumors in 9 cases observed for a mean time of 44 months (range 12 to 86), the mean increase in tumor size was 0. 531 cm/year. There were 19(65.5%) tumors>3 cm at the end of follow-up but only 1 developed retroperitoneum lymph nodes metastasis. A total of 87 solid tumors were removed and 62 (71.3%) solid tumors were man-aged by NSS. Pathological results showed 86 clear cell carcinomas (73 Fuhrman Ⅰ and 12 Fuhrman Ⅱ ) and 1 calcified lesion. During mean follow-up of 50(5-237) months, local recurrence occurred in 4 cases treated with NSS; 26 patients were alive at the end of follow-up. Conclusions DNA testing might be helpful in the earlier detection of asymptomatic VHL patients. Most solid renal tumors in VHL disease grow slowly. The majority of the tumors >3 cm may still be indolent and do not metas-tasize during longer follow-up and can be observed. NSS is effective and safe for RCC in VHL disease.

To develop an analytic method for qualitative and quantitative analysis of dodecatetraenamides A, B in 42 samples of two official species of Asari Radix et Rhizoma( ARR) (37 samples of Asarum heterotropoides var. mandshuricum with different collection time and 5 samples of Asarum sieboldiivar. seoulense). The HPLC-IT-TOF-MS/MS methods for the qualitative and UPLC-PDA methods for the quantitative analysis were established. Dodecatetraenamides A, B were identified by comparing the retention time, UV absorption spectrum and quasi-molecular ion peak [ M + H]+ with the reference compound using HPLC-IT-TOF-MS/MS. The content of dodecatetraenamides A and B in ARR were determined by UPLC-PDA. The separation was successfully carried out on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 µm) column eluted with mobile phases of water (A) and acetonitrile (B) in gradient program (0-3 min, 35% B; 3-5 min, 35%-36% B; 5-6 min, 36%-43% B; 6 min-11 min 43% B; 11-12 min, 43%-100% B). The column temperature was 45 °C, and the detection wavelength was set at 254 nm. The flow rate was 0.6 mL · min(-1). On one level mass spectrometry scanning, the results showed that the quasi-molecular ion [M + H] + of both dodecatetraenamides A and B were m/z 248.20. The quantitative method with UPLC-PDA has made the baseline separation of the constituents, which were reported as mixtures in the most literatures. The average recovery of dodecatetraenamides A and B were 97.90% and 99.86%, the relative standard deviation were 0.4% and 1.1%, respectively. The contents of dodecatetraenamides A, B in all ARR samples was in the range of 0.11-3.89 and 0.24-6.65 mg · g(-1). Their contents reduced with the extension of storage time. Compared with the samples of 2013, the average content of the two constituents in the samples collected in year 2002-2003 reduced 34% and 36%, respectively (P < 0.05). Compared the A. sieboldii var. seoulense and A. heterotropoides var. mandshuricum with the same collective time and production area, the average contents of the two constituents in latter were up to (1.59 ± 0.75) mg · g(-1) and (2.90 ± 1.17) mg · g(-1), respectively, significantly higher than that in A. sieboldii var. seoulense (dodecatetraenamide A were (0.78 ± 0.52) mg · g(-1), dodecatetraenamide B were (1.69 ± 0.83) mg · g(-1)) (P < 0.05). The content of the dodecatetraenamide A in overground part was in the range of 0.11-0.33 mg · g(-1), dodecatetraenamide B was 0. 24-0.60 mg · g(-1), which were much lower than that of the underground part of ARR (dodecatetraenamide A was in the range of 0.73-3.89 mg · g(-1), dodecatetraenamide B was 2.11-6.24 mg · g(-1)). The method was certified to be simple, accurate and reliable and could be used for qualitative and quantitative analysis of dodecatetraenamide A and B in different species of ARR, also can be used for the comprehensive quality control of traditional Chinese medicine, Asari Radix et Rhizoma.
Amides ; chemistry ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Rhizome ; chemistry

Objective To explore the effects and safety of hemoperfusion(HP) therapy on tetramine poisoning in infants.Methods Thirty-five infants with tetramine poisoning were divided into two groups: HP group(n=18) and non HP group(n=17).The changes of blood tetramine concentration and clinical symptom improving of both groups after the treatment were observed together with the adverse effects of HP group.Results The average blood tetramine concentration of HP group was higher than that of non HP group(342.2?333.4 vs 117.9?50.8 ?g/L,P

Objective To assess the prognosis of patients with bladder neck (BN) involvement in radical prostatectomy specimens and compare it with patients with seminal vesicle invasion (SVI).Methods From January 2002 to December 2008,42 patients with prostate carcinoma who underwent radical prostatectomy were analyzed retrospectively.The mean age was 70.3 years (range 59-78 years).The patients were divided into two groups according to 2002 TNM system:pT4a group (bladder neck invasion) with 17 cases and pT3b group (unilateral or bilateral seminal vesicle invasion)with 25 cases.Postoperative outpatients were followed-up on a regular basis,two consecutive postoperative serums PSA greater than 0.2 μg/L defined as a biochemical recurrence.The postoperative follow-up to the time of biochemical recurrence or follow up to the deadline without biochemical recurrence were defined as biochemical recurrence free survival time.Results All patients were followed up for 50-122 months.The patients with BN involvement 17.6％ (3/17) had biochemical recurrence,whereas seminal vesicle invasion 24.0％ (6/25),there was no statistical significance between the two groups (P＞0.05).Conclusions Postoperative adjuvant therapies can improve the biochemical recurrence free survival.In this study,the prognosis of pT4a and pT3b group is similar.A conceivable down-staging of BN involvement in the TNM staging system should be considered.

Amyloid beta-Protein Precursor ; Angiotensin II ; Anti-Inflammatory Agents/pharmacology* ; Antioxidants/pharmacology* ; HaCaT Cells ; Humans