Survivin is a protein that inhibits apoptosis and regulates cell division.The cDNA sequence of survivin was amplified by RT-PCR and sub-cloned into the prokaryotic expression vector pET-21b(+),followed by transformation into E.coli strain BL21(DE3) and induction with IPTG.The recombinant survivin protein fusing with 6?His tag was expressed in E.coli in the form of inclusion body at the expression level over 60% of the total cell protein.Results of Western blotting showed that recombinant survivin reacted specifically with anti-human survivin antibody.After gel filtration,the recombinant protein reached the purity over 95%,which facilitate the study of diagnosing and inhibitor agents targeting survivin.

Twelve compounds were isolated from the venom of Bufo bufo gargarizans. On the basis of their physical and chemical properties and spectral data, their structures were identified as resibufagenin (1), bufotalin (2), desacetylcinobufagin (3), 19-oxodesacetylcinobufotalin (4), cinobufotalin (5), 1beta-hydroxylbufalin (6), 12alpha-hydroxybufalin (7), bufotalinin (8), Hellebrigenin (9), telocinobufagin (10), hellebrigenol (11) and cinobufagin-3-hemisuberate methyl ester (12), respectively. Compounds 7 and 12 are new natural products.
Animals ; Bufanolides ; chemistry ; Bufo bufo ; Medicine, Chinese Traditional ; Molecular Structure ; Venoms ; chemistry

OBJECTIVETo compare the effects and complications of subinguinal microscopic ligation, laparoscopic transperitoneal varicocelectomy, laparoscopic retroperitoneal varicocelectomy, open retroperitoneal high ligation, and interventional embolotherapy in the treatment of varicocele.

METHODSWe conducted a retrospective study that included 632 varicocele patients treated by subinguinal microscopic ligation (group A, n = 79), laparoscopic transperitoneal varicocelectomy (group B, n = 120), laparoscopic retroperitoneal varicocelectomy (group C, n =137), open retroperitoneal high ligation (group D, n = 283), and interventional embolotherapy (group E, n = 13). We compared the baseline and 3-month postoperative semen parameters, postoperative complications, and pregnancy rate among the five groups of patients.

RESULTSThe operation time was longer in groups A ([2.02 ± 1.25] h) and E ([2.17 ± 1.02] h) than in the other three groups, while the postoperative hospital stay was the shortest in group E ([1.1 ± 0.1] d). Intestinal injury or incision bleeding occurred intraoperatively in 2 cases in group B and 1 case in group E. Postoperative scrotal edema developed in 3.7, 17, 10, and 19% of the patients in groups A, B, C, and D, respectively, but not in group E. The rate of 1-year recurrence was the lowest in group A (1.6%) and highest in group E (22%). Sperm concentration and the percentages of progressively motile sperm and morphologically normal sperm were improved postoperatively in all the patients (P < 0. 05), but there were no statistically significant differences among the five groups either in the above three parameters or in the postoperative pregnancy rate (P > 0. 05).

CONCLUSIONIn the surgical treatment of varicocele, laparoscopic retroperitoneal approach involves short operation time and few complications, subinguinal microscopic ligation has the advantages of little injury, rapid recovery, and few complications but requires specialized microsurgical techniques, and interventional embolotherapy leaves no incision scar and needs only local anesthesia and 1-day postoperative hospital stay, which is uitable for those with a contraindication to anesthesia.

Embolization, Therapeutic ; adverse effects ; methods ; Female ; Humans ; Laparoscopy ; adverse effects ; methods ; Length of Stay ; Ligation ; adverse effects ; methods ; Male ; Operative Time ; Postoperative Complications ; Postoperative Hemorrhage ; Pregnancy ; Pregnancy Rate ; Recurrence ; Retroperitoneal Space ; Retrospective Studies ; Sperm Count ; Urogenital Surgical Procedures ; adverse effects ; Varicocele ; surgery ; Vascular Surgical Procedures ; adverse effects ; methods

OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.

METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.

RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.

CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.

Animals ; Autophagy ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats

Anesthesia ; Fiber Optic Technology ; Humans ; Intubation, Intratracheal

OBJECTIVETo study the impact of left varicocele (VC) and varicocelectomy (VCT) on the apoptosis of spermatogenic cells and the levels of nitrogen monoxidum (NO) and interleukin 1 (IL-1) in the rat testis.

METHODSWe randomly divided 60 adolescent male SD rats into four groups of equal number: sham operation control, VC model 1 (VC1), VC model 2 (VC2), and VCT. We determined the semen quality and levels of NO and IL-1 in the testis tissue, detected the apoptosis of spermatogenic cells by TUNEL, and compared the indexes obtained among different groups.

RESULTSAn experimental VC model was successfully established by partially ligating the left renal vein of the rats. Sperm concentration and motility were significantly decreased in the VC1 ([1.54 ± 1.16] x 10⁶/ml and [44.23 ± 15.46]%) as compared with those in the sham operation group ([2.80 ± 1.62] x 10⁶/ml and [72.34 ± 12.62]%) (P < 0.05), but remarkably higher in the VCT ([1.82 ± 1.34] x 10⁶/mI and [51.21 ± 12.62]%) than in the VC2 group ([1.04 ± 1.21] x 10⁶/ml and [39.23 ± 13.21]%) (P < 0.05). The levels of NO and IL-1 in the left testes were markedly elevated in the VC1 ([0.172 ± 0.030] ng/ml and [1.468 ± 0.080 ] mg/ml) in comparison with those in the sham operation group ([0.134 ± 0.021] ng/ml and [0.782 ± 0.079 ] mg/ml) (P < 0.05), and significantly higher in the VC2 ([0.198 ± 0.020] ng/ml and [1.994 ± 0.090] mg/ml) than in the VCT group ([0.141 ± 0.010] ng/ml and [0.781 ± 0.036] mg/ml) (P < 0.05). However, the NO and IL-1 levels in the right testis showed no statistically significant differences between the two groups, and the two levels were positively correlated (r = 0.492, P < 0.01). The rats of the VC1 group exhibited remarkable apoptosis of spermatogenic cells in the bilateral testes, with significant differences in the apoptosis index ( AL) between the two sides (P < 0.05) as well as in the same side in comparison with the sham operation group (P < 0.01). The Als of spermatogenic cells in the bilateral testes showed statistically significant differences in the VCT (P < 0.05) but not in the VC2 group (P > 0.05), and those in the same side manifested dramatic differences between the VCT and VC2 groups (P < 0.01).

CONCLUSIONVaricocele induces changes of the NO and IL-1 levels in the testis tissue and increases the apoptosis of spermatogenic cells, which might be one of the causes of testis damage and spermatogenic dysfunction.

Animals ; Apoptosis ; Germ Cells ; pathology ; In Situ Nick-End Labeling ; Interleukin-1 ; analysis ; Ligation ; Male ; Nitrogen ; analysis ; Random Allocation ; Rats ; Semen Analysis ; Spermatogenesis ; Testis ; chemistry ; Varicocele ; complications ; surgery

ObjectiveTo investigate the role of platelet-derived growth factor(PDGF) and its receptor in the development of hypertrophic scar. MethodsThe expression of PDGF and its receptor were detected in biopsy specimens of 9 pieces of normal skin, 7 pieces of granulation tissue of burn wound and 34 pieces of hypertrophic scar by immunohistochemical staining using specific polyclonal antibodies.ResultsPDGF and its receptor markedly increased in granulation tissue and hypertrophic scars, reaching the peak in the hypertrophic scars within 6 months and then decreased after the peak, whereas PDGF and its receptor expressed weakly in only a few normal skin specimens, and the differences were significant(P<0.05).ConclusionsThe increasing expression of PDGF and its receptor may be related to the development of hypertrophic scar.

BACKGROUNDAcute renal failure following coronary artery bypass grafting (CABG) surgery is associated with high morbidity and mortality. Approximately half of all patients who develop acute kidney injury (AKI) subsequently develop acute renal failure (ARF). The purpose of the study was to compare early transient changes in renal function within the first post-operative week following CABG in patients that were either off-pump or on-pump.

METHODSEight hundred and forty-nine consecutive patients with isolated CABG in a single institution between January 1990 and August 2006 were retrospectively analyzed, including 518 off-pump and 331 on-pump patients. A multivariate Logistic regression model was constructed to identify risk factors for the development of AKI.

RESULTSSixty-one off-pump patients and 63 on-pump patients developed AKI. Risk factors for the development of post-operative AKI included an ejection fraction > or =50% or < or =30%, a pulse pressure > or =60 mmHg, peripheral vascular disease, diabetes, emergent procedure, triple-vessel disease, body mass index, peri-operative and post-operative. intra-aortic balloon pumping, NYHA class III and IV, and cardiopulmonary bypass. An ejection fraction > or =50% and peri-operative and post-operative intra-aortic balloon pumping were protective (OR <1). Peak serum creatinine for post-operative AKI was noted 12 hours and 24 hours in the off-pump and on-pump patients, respectively. Serum creatinine kinetics revealed rapid recovery in the 24th to 48th hour (off-pump) and the 48th to 72nd hour (on-pump).

CONCLUSIONRenal protection strategies are indicated from general anesthesia induction until 48 and 72 hours post-operatively in off-pump and on-pump patients, respectively.

Acute Kidney Injury ; etiology ; Aged ; Coronary Artery Bypass ; adverse effects ; Coronary Artery Bypass, Off-Pump ; adverse effects ; Creatinine ; blood ; Female ; Humans ; Kidney ; physiopathology ; Male ; Middle Aged ; Retrospective Studies

Design space approach is applied in this study to enhance the robustness of first ethanol precipitation process of Codonopsis Radix (Dangshen) by optimizing parameters. Total flavonoid recovery, dry matter removal, and pigment removal were defined as the process critical quality attributes (CQAs). Plackett-Burman designed experiments were carried out to find the critical process parameters (CPPs). Dry matter content of concentrated extract (DMCE), mass ratio of ethanol to concentrated extract (E/C ratio) and concentration of ethanol (CEA) were identified as the CPPs. Box-Behnken designed experiments were performed to establish the quantitative models between CPPs and CQAs. Probability based design space was obtained and verified using Monte-Carlo simulation method. According to the verification results, the robustness of first ethanol precipitation process of Dangshen can be guaranteed by operating within the design space parameters. Recommended normal operation space are as follows: dry matter content of concentrated extract of 45.0% - 48.0%, E/C ratio of 2.48-2.80 g x g(-1), and the concentration of ethanol of 92.0% - 92.7%.
Chemical Precipitation ; Chemistry, Pharmaceutical ; methods ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification