Objective To investigate the protective effects and mechanism of shenmai injection(SMI) on cardiomyocytes injured by hydrogen peroxide(H 2O 2) or angiotension Ⅱ(AngⅡ). Methods According to the different concentrations of H 2O 2, AngⅡ,SMI,Vitamin C(VitC) added into the cardiomyocytes culture media(CCM), the cultures were divided into 15 groups. Lactate dehydrogenase(LDH ),malondialdehyde(MDA),superoxide dismutase (SOD) of CCM and cardiomyocytes viability(CMV), Na +-K +-ATPase,Ca 2+-ATPase?cardiomyocytes apoptosis rate(CMAR) ,nitrogen oxide(NO), nitrogen oxide synthesis enzyme(NOS),eNOS expression of cardiomyocytes were detected respectively .Results H 2O 2 or AngⅡ could decrease CMV, SOD, Na +-K +-ATPase,Ca 2+-ATPase,NOS,NO,eNOS expression and increase CMAR,LDH and MDA.SMI could lessen the changes of the items mentioned above ,which were caused by H 2O 2 or AngⅡ.The effects of SMI 10 ml/L were stronger than those of SMI 5 ml/L or VitC 50 mg/L.Conclusion SMI has a significant protective effect on cardiomyocyte injured by H 2O 2 or AngⅡ.

OBJECTIVETo understand the characteristics of pncA gene in multidrug-resistant Mycobacterium tuberculosis isolates and its correlation with drug resistance to pyrazinamide.

METHODSA total of 127 clinical isolates of multidrug-resistant mycobacterium tuberculosis were collected from Shenzhen from year 2007 to 2009. PZA susceptibility was determined by the BACTEC MGIT 960 PZA method. Pyrazinamidase (PZase) activity testing and pncA gene sequencing were performed in all the isolates. The type and frequency of mutations in pncA were determined. Correlation analysis among PZA susceptibility and PZase activity, pncA mutation was performed.

RESULTSAmong the 127 isolates, 62 isolates (48.8%) were found resistance to PZA. Among the 62 PZA resistant isolates, 45 isolates which had various pncA mutations were negative for PZase. Mutation rate was 77.4% (48/62) in total PZA resistance isolates. Different types of 48 resistant isolates were identified in the pncA gene, including base substitution (33 isolates), frame shift mutation (12 isolates) and codon mutation (3 isolates). No mutations except one isolate (N11D) existed in all PZA-susceptibility isolates which were positive for PZase. A total of 5 mutations which have not been described previously were found as follows: H57P, P62Q, G108R, D110Y and G162V. The correlation among the PZA susceptibility and the PZase activity (r = 0.895, P < 0.05), the pncA mutation (r = 0.779, P < 0.05) were significant in 127 multidrug-resistant isolates.

CONCLUSIONA high diversity of pncA gene mutation was found among PZA resistant strains of MTB. This study revealed five new mutations of the pncA gene that were not previously described, which scattered in the hot-spot regions located in the metal coordination site and active site of the enzyme. Mutations had a high correlation with the PZA resistance.

Antitubercular Agents ; pharmacology ; DNA, Bacterial ; genetics ; Humans ; Mutation ; Mycobacterium tuberculosis ; drug effects ; genetics ; isolation & purification ; Pyrazinamide ; pharmacology ; Tuberculosis, Multidrug-Resistant ; genetics ; microbiology

A series of aralkyl-ketone-4-piperidol derivatives were synthesized and tested for their analgesic activities. All of the novel 30 compounds were prepared from 4-piperidone and alpha-halo-aralkyl-ketone through five steps, including Boc protection, nucleophilic addition in presence of CeCl3/NaI catalyst, deprotection, condensation and salification. Their structures were confirmed by 1H NMR and HRMS. Preliminary in vivo pharmacological trials showed that most of the synthesized compounds revealed analgesic effects. Among the tested compounds, 8, 13 and 22 exhibited potent analgesic activities in both mice writhing and mice hot plate model. The three compounds have low affinity for mu, delta, kappa receptors, which is a chance to find a better precursor of non-opioid analgesic for further optimization.
Analgesics, Non-Narcotic ; chemical synthesis ; chemistry ; pharmacology ; Animals ; Mice ; Molecular Structure ; Pain Measurement ; Pain Threshold ; drug effects ; Piperidones ; chemical synthesis ; chemistry ; pharmacology ; Receptors, Opioid, delta ; metabolism ; Receptors, Opioid, kappa ; metabolism ; Receptors, Opioid, mu ; metabolism ; Structure-Activity Relationship

ObjectiveTo compare the effects of single sperm cryopreservation and conventional cryopreservation on embryo culture and pregnancy in patients undergoing microdissection testicular sperm extraction （micro-TESE）. MethodsA retrospective analysis was done on the patients who underwent micro-TESE at the Reproductive Medicine Center in the Sixth Affiliated Hospital of Sun Yat-Sen University between January 2018 and December 2021. The single sperm cryopreservation group included 39 patients undergoing single sperm cryopreservation and 307 MII oocytes. The conventional cryopreservation group included 39 patients undergoing conventional cryopreservation and 410 MII oocytes. Propensity score matching （PSM） was performed to balance the selection bias. The fertility rate， embryo culture and pregnancy of these two groups were compared. ResultsThere was no statistical difference in age， body mass index （BMI）， years of infertility， basal FSH， basal LH， basal E2， AMH， AFC， number of oocytes retrieved and number of transferred embryos between the two groups （P>0.05）. No significant difference was found in fertilization rate （65.8% vs. 60.49%）， available embryo rate （67.82% vs. 58.87%） and high-quality embryo rate （70.80% vs. 75.34%）. The single sperm cryopreservation group had significantly lower clinical pregnancy rate than conventional cryopreservation group （45.45% vs. 70.0%， P=0.049）. ConclusionIf the patients undergoing micro-TESE have very few sperms， single sperm cryopreservation could be an effective choice to increase the utilization of each sperm and ensure the subsequent sperm retrieval after thawing， but the clinical pregnancy rate is decreased. Due to the limited number of cases， we need a large additional number of cases to observe and analyze.

OBJECTIVEThis paper presents the experience in using the reverse buccinator musculomucosal flap for repairing the defect following excising inverted papilloma of the nose.

METHODSAfter the inverted papilloma of the nose was excised through an endonasal approach, the reverse buccinator musculomucosal flap supplied by the retrograde blood flow of the anterior buccal artery was harvested and sutured to the defect through the ora-nasal tunnel. The procedure was performed on three patients.

RESULTSThe postoperative course was uneventful. All the flaps survived completely.

CONCLUSIONSThe technique provides the solution to prevent nasal stricture from cicatricial contracture after excising inverted papilloma. In the operation, excising the inverted papilloma and repairing the defect was performed simultaneously, saving another operation for the secondary deformity. The technique is also applicable to the treatment of existing cicatricial stricture of the nose.

Humans ; Male ; Middle Aged ; Mouth Mucosa ; transplantation ; Nose Neoplasms ; pathology ; surgery ; Papilloma, Inverted ; pathology ; surgery ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps

Objective Serum total prostatic specific antigen (tPSA), free prostatic specific antigen (fPSA), fPSA/tPSA ratio, and prostate cancer-specific antigen density (PSAD) were determined to explore the best identification point, thus improving the specificity of early screening of prostate cancer. Methods The tPSA, fPSA, fPSA/tPSA, and PSAD of patients with benign prostatic hyperplasia group (n=250) and prostate cancer group (n=92) were tested, and the receiver operating characteristic (ROC) curve was drawn to determine the best cutoff value for the evaluation. Results When the cutoff values of tPSA, fPSA/tPSA, and PSAD were at 11.3 mg/L, 0.16, and 0.18 mg/(L·cm³), respectively, the specificity and sensitivity were the best:82.4% and 84.2% for tPSA, 76.9% and 81.7% for fPSA/tPSA, and 83.1% and 80.4% for PSAD.When the best cutoff values of tPSA, fPSA/tPSA, and PSAD were combined in analysis, the specificity and sensitivity of fPSA/tPSA and PSAD combination showed the best result (92.4% and 81.4%, respectively).When the tPSA value was in the range of 4-10 mg/L, the optimal cutoff values of PSAD and fPSA/tPSA were 0.21 mg/(L·cm³) and 0.15, and the specificity and sensitivity reach the best, which were 84.1% and 80.1%, 81.0 % and 80.3%, respectively. Conclusion Combination of tPSA, fPSA/tPSA and PSAD analysis is better than any single of them in early screening of prostate cancer.The specificity and sensitivity of combined use of fPSA/tPSA and PSAD could serve as an optimal screening reference value.

OBJECTIVETo study the effects of Rapamycin (RPM) on angiogenesis and tumor progression in human hepatocellular carcinoma (HCC) implantation mice.

METHODSTumor tissues of HCC were implanted into the liver of nude mice. Then, nude mice were treated with RPM and cyclosporine A (CsA). Real-time PCR was used to detect the mRNA expression of vascular endothelial growth factor (VEGF). Immunohistochemical stain and image analysis were used to detect the protein expression of VEGF and proliferating cell nuclear antigen (PCNA) and microvessel density (MVD) was counted by endothelial cells immunostained by anti-CD34 antibody. The concentration of VEGF in the peripheral blood was detected by ELISA.

RESULTS(1) The tumor weights were (372 +/- 35) mg, (769 +/- 39) mg and (751 +/- 42) mg in RPM, CsA and control group respectively. The tumor weight was significantly decreased in RPM group and no difference in CsA group compared with control group. (2) The expression of VEGF mRNA, VEGF and PCNA protein in tumor tissues and concentration of VEGF in the peripheral blood were remarkably down-regulated in RPM group compared with control group (P < 0.05) and were not remarkably different in CsA group from in control (P > 0.05).(3) Comparing with the control, the tumor MVD was remarkably decreased in RPM group (P < 0.05), and no difference in CsA group (P > 0.05).

CONCLUSIONRPM can inhibit angiogenesis and tumor progression of HCC by down-regulated the gene and protein expression of VEGF and inhibited the growth of tumor.

Animals ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Humans ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Mice ; Mice, Nude ; Neovascularization, Pathologic ; drug therapy ; Proliferating Cell Nuclear Antigen ; metabolism ; RNA, Messenger ; genetics ; Sirolimus ; pharmacology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Xenograft Model Antitumor Assays

OBJECTIVETo observe the effects of Qiangjing Tablets (QJT) on the semen quality and Fas/FasL signaling pathway in male SD rats with infertility.

METHODSModels of infertility were made in 50 male SD rats by intragastric administration of Tripterygium (GTW) for 3 weeks, and another 20 rats were taken as blank controls. Then 40 successfully established rat models were randomly divided into four groups, model control, low-dose QJT, medium-dose QJT, and high-dose QJT, the latter three groups treated intragastrically with QJT at 58 mg, 105 mg, and 233 mg per kg of the body weight per day, respectively. After 4 weeks of medication, the rats were killed for examination of semen quality and determination of the expression of the apoptosis factor FasL in the testis tissue.

RESULTSCompared with the blank controls, the model rats showed significant decreases in sperm concentration ([71.99 ± 9.72] vs [10.94 ± 3.58] x 10⁶/ml, P < 0.01), motility ([48.95 ± 4.10] vs [9.31 ± 5.79]%, P < 0.01), and viability ( [82.06 ± 6.16] vs [24.03 ± 6.93]%, P < 0.01). In comparison with the model controls, the rats in the QJT groups exhibited remarkably increased sperm concentration, motility, and viability, more significantly in the high-dose group ([59.66 ± 4.53] x 10⁶/ml, [35.45 ± 5.21] %, and [61.97 ± 9.75]%) and medium-dose group ([40.89 ± 4.90] x 10⁶/ml, [24.41 ± 4.79]%, and [60.06 ± 10.62]%) (P < 0.05 or P < 0.01). The expression of FasL was markedly reduced in the low-, medium-, and high-dose QJT groups (0.5215 ± 0.0189, 0.5371 ± 0.0364, and 0.4556 ± 0.0215) as compared with that of the model controls (0.5989 ± 0.0448 ) (P < 0.05 or P < 0.01).

CONCLUSIONBy upregulating the Fas/FasL signaling pathway, Tripterygium glycosides may induce the apoptosis of spermatogenic cells and reduce sperm concentration, motility and viability, resulting in infertility. The Chinese medicine Qiangjing Tablets can improve the reproductive function of male rats by decreasing the expression of the apoptosis factor FasL in the testis.

Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fas Ligand Protein ; drug effects ; metabolism ; Germ Cells ; Glycosides ; Infertility, Male ; chemically induced ; drug therapy ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Semen ; drug effects ; Semen Analysis ; Signal Transduction ; Sperm Count ; Sperm Motility ; drug effects ; Tablets ; Testis ; drug effects ; metabolism ; Tripterygium

Animals ; Cell Transformation, Neoplastic ; Cloning, Molecular ; Hepacivirus ; genetics ; immunology ; pathogenicity ; Hepatitis C Antigens ; toxicity ; Liver Neoplasms, Experimental ; pathology ; virology ; Male ; Oncogenic Viruses ; pathogenicity ; RNA, Messenger ; toxicity ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; toxicity ; Viral Core Proteins ; toxicity ; Virion ; pathogenicity

BACKGROUNDCell-based vascular therapies of endothelial progenitor cells (EPCs) mediated neovascularization is still a novel but promising approach for the treatment of ischemic disease. The present study was designed to investigate the therapeutic potentials of human umbilical cord blood-derived EPCs (hUCB-EPCs) in rat with acute myocardial infarction.

METHODSHuman umbilical cord blood (hUCB) mononuclear cells were isolated using density gradient centrifugation from the fresh human umbilical cord in healthy delivery woman, and cultured in M199 medium for 7 days. The EPCs were identified by double-positive staining with 1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethylindocarbocyanine percholorate-labeled acetylated low-density lipoprotein (Dil-Ac-LDL) and fluorescein isothiocyanate-conjugated Ulex europaeus lectin (FITC-UEA-l). The rat acute myocardial infarction model was established by the ligation of the left anterior descending artery. The hUCB-EPCs were intramyocardially injected into the peri-infarct area. Four weeks later, left ventricular function was assessed by a pressure-volume catheter. The average capillary density (CAD) was evaluated by anti-VIII immunohistochemistry staining to reflect the development of neovascularization at the peri-infarct area. The graft cells were identified by double immunofluorescence staining with human nuclear antigen (HNA) and CD31 antibody, representing human origin of EPCs and vascular endothelium, respectively. Expressions of cytokines, proliferating cell nuclear angigen (PCNA), platelet endothelial cell adhesion molecule (PECAM) and vascular endothelial growth factor (VEGF) were detected to investigate the underlying mechanisms of cell differentiation and revascularization.

RESULTSThe donor EPCs were detectable and integrated into the host myocardium as confirmed by double-positive immunofluorescence staining with HNA and CD31. And the anti-VIII staining demonstrated a higher degree of microvessel formation in EPCs transplanted rats, associated with a significant improvement of global heart function in terms of the increase of left ventricular end-systolic pressure (LVESP), +dp/dtmax and -dp/dtmax as well as the decrease of LVEDP in rats with EPCs therapy comparing to the control rats (P < 0.05). Moreover, the expression of the rat PCNA mRNA and PECAM were both enhanced in the EPCs group compared with that of the control group.

CONCLUSIONSThe human umbilical cord blood-derived EPCs could incorporate into new-born capillaries in rat myocardium, induce revascularization and improve the proliferation activity in the peri-infarct area, resulting in the improvement of global heart function. This may indicate a promising stem cell resource in cell-based therapy for ischaemic diseases.

Animals ; Cells, Cultured ; Cytokines ; metabolism ; Endothelial Cells ; cytology ; physiology ; Endothelium, Vascular ; Fluorescent Antibody Technique ; Humans ; Male ; Myocardial Infarction ; metabolism ; therapy ; Neovascularization, Physiologic ; physiology ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; Rats ; Rats, Wistar ; Stem Cell Transplantation ; Stem Cells ; cytology ; Umbilical Cord ; cytology ; Vascular Endothelial Growth Factor A ; metabolism