Objective:To design Keap1-tat peptide and explore its neuroprotective role on hipocampal CA1 neuron,as well as the effect on spacial learning and memory function following global cerebral ische-mia.Methods:Adult male Sprague Dawley (SD)rats were subjected to global cerebral ischemia (GCI) by four-vessel occlusion for 1 5 min and randomly divided into five groups:sham,sham+Keap1-tat,is-chemia/reperfusion (I/R),Keap1-tat peptide-and vehicle-administrated groups.For Keap1-tat or vehi-cle groups,the rats were treated with Keap1-tat (30,50,1 00 μg in 5 μL 0.9%saline)or the same vo-lume vehicle by intracerebroventricular injection (icv)30 min prior to ischemia.Cresyl violet staining was used to observe the surviving neurons and 4-hydroxy-2-noneal (4-HNE ) and 8-hydroxy-2′-deox-yguanosine (8-OHdG)immunostaining were used to detect the change of markers response to oxidative stress in hippocampal CA1 region.The spatial learning and memory function of the rats was evaluated using Morris water maze.Results:Compared with sham group,the number of surviving neurons in ische-mia-reperfusion and vehicle groups significantly decreased in the hippocampal CA1 region (P<0.05 ), while administration of Keap1-tat significantly decreased the damage following GCI (P<0.05),and the dose of 50 μg existed the most effective neuroprotective role.Furthermore,immunostaining intensity of 4-HNE and 8-OHdG,markers of oxidative stress damage attenuated by Keap1-tat peptide as compared with vehicle group in CA1 region.Of significant interest,the time of finding underwater platform in Keap1-tat group animals was significantly short,and after removing the platform,the probe time of Keap1-tat group animals in the original quadrant where the platform was significantly increased compared with that of vehi-cle and I/R group animals (P<0.05).Conclusion:Keap1-tat peptide can effectively attenuate neuro-nal damage in hippocampal CA1 region and improve learning and memory function,which might bedue to the attenuation of oxidative stress caused by GCI.

The stability of melittin in different solvents (water, deoxygenated water, physiological saline, PBS, 50% ethanol, ethanol, glycerol)was studied and the results showed that the stability of melittin is not influenced by light, temperature and pH in 50% ethanol, which melittin can be completed dissolved when compared with ethanol and glycerol, in such, 50% ethanol was chosen as solvent storage when measured content of melittin. Then the effect of different concentrations of PBS, the pH of PBS and rat skin ho- mogenates were tested, and the results showed that melittin was degraded rapidly at low concentration solution and low ionic strength. Increasing pH of PBS and rat skin homogenate can accelerate the degradation of melittin. These researches provide an experimental ba- sis for further study of melittin.
Animals ; Drug Stability ; Ethanol ; chemistry ; Hydrogen-Ion Concentration ; Melitten ; chemistry ; Rats ; Skin ; drug effects ; Solvents ; chemistry ; Temperature

OBJECTIVETo compare the effect of transumbilical single-site single-port with that of transumbilical single-site double-port laparoscopic varicocelectomy in the treatment of varicocele in adolescents.

METHODSWe randomly assigned 80 varicocele patients aged 10 - 16 years to two groups of equal number to receive transumbilical single-site single-port and single-site double-port laparoscopic varicocelectomy, respectively. We compared the operation time, postoperative hospital stay, incisional pain, complications and satisfaction with the abdominal cosmetic outcomes between the two groups.

RESULTSAll the operations were successfully performed. The double-port group showed a significantly higher score on the Visual Analogue Scale than the single-port group (4.8 +/- 1.4 vs 3.6 +/- 1.1, t = -4.986, P < 0.01), but there were no significant differences between the two groups in the operation time ([29.8 +/- 4.2] vs [31.2 +/- 4.6] min, t = 1.383, P = 0.171), postoperative hospital stay ([1.95 +/- 0.7] vs [1.82 +/- 0.8] d, t = -0.784, P = 0.436), complications (0 vs 0) and scores on the satisfaction with abdominal cosmetic outcomes (4.6 +/- 0.6 vs 4.8 +/- 0.5, t = 1.253, P = 0.214). No recurrence, umbilical hernia, hydrocele and orchiatrophy were found in the two groups of patients at 6 months after operation, and no visible scar was observed on the abdominal surface.

CONCLUSIONWith strict surgical indications, single-site single-port and single-site double-port laparoscopic varicocelectomies have similar clinical effects in the treatment of varicocele, which leave no scar on the abdominal surface. Single-site double-port laparoscopy needs no special instruments and therefore is worthier of wide clinical application.

Adolescent ; Child ; Humans ; Laparoscopy ; methods ; Length of Stay ; Male ; Operative Time ; Umbilicus ; surgery ; Varicocele ; surgery

ObjectiveTo evaluate the improvement on test performance of UniCel DxH 800 automated hematology analyzer for complete blood count (CBC) by detecting its performance indicators and comparing the differences of the results with LH 750 hematology analyzer and ADVIA 2120 hematology analyzer.MethodsThe precision,carryover and linearity of UniCel DxH 800 in measurement of CBC were evaluated by using fresh blood samples and instrument quality control of products.To evaluate the accuracy of leukocyte differential count and reticulocyte count with the microscopic method as the “gold standard”.To calculate the bias and correlation between the results measured by LH 750,ADVIA 2120 and UniCel DxH 800 hematology analyzers and compare these three instruments on the validity of the alarm in abnormal cells.ResultsIntra-precision:The coefficient of variation (CV) of the results of RBC,Hb and MCV were less than 0.5％,the CV of WBC and PLT results were less than 1.5％.Inter-precision:the CV of the parameters above were less than 2.5％.The carryover rate of WBC,RBC,Hb,MCV and PLT were less than 0.51％.In the concentration range covered by clinical samples,the correlation coefficients between the measured values and theoretical value in testing WBC,RBC,Hb and PLT were greater than 0.999 ( P ＜0.01 ).The measurement results of WBC,RBC,Hb,MCV and PLT hy UniCel DxH 800,ADV1A 2120 and LH 750 hematology analyzers have good correlation (r ＞ 0.973,P ＜ 0.01 ).Correlation of reticulocyte count between the UniCel DxH 800 hematology analyzer and microscolpic method was significant (r =0.920,P ＜0.01 ).Correlation of leukocyte differential count about the grauulocytes, lymphocytes and eosinophils between the UniCel DxH 800 hematology analyzer and microscopic method was good (r =0.914,0.900 and 0.725,P ＜0.01 ),followed by monocytes ( r =0.612,P ＜0.01 ),which were better than the LH 750 with similar detection principle.The UniCel DxH 800 hematology analyzer demonstrated higher sensitivity (96.6％ ) for the alarm of abnormal cells and achieved a lower false-negative rate (2.5％ ).Meanwhile,the sensitivity of the neutrophil nuclei left shift was higher (90.5％ ) and the false-negative rate (5.0％) was lower.ConclusionsThe UniCel DxH 800 hematology analyzer for complete blood count shows advantages of high precision,low carryover rate and wide linear range.The results detected by the UniCel DxH 800 hematology analyzer have good correlation with the LH 750 and ADVIA 2120.

Research on polymer impurities has always been important in the quality control of cephalosporins. Research on polymers in cephalosporins that lack active amino groups on the C-7 side chain has not been reported. Therefore, our study used cefazolin sodium, which is widely used in the clinic, as an example. The polymer in cefazolin sodium and its product "cefazolin sodium pentahydrate for injection" was analyzed by column switching liquid chromatography-high resolution mass spectrometry. Two polymer impurity peaks were detected and the possible structures of these polymers were suggested. Through two-dimensional liquid chromatography, the chromatographic peaks following Sephadex gel chromatography and high-performance gel chromatography were compared to those obtained by reverse high-performance liquid chromatography (HPLC) for cefazolin sodium as reported in the Chinese Pharmacopoeia. The HPLC method proves more suitable for polymer detection than Sephadex gel chromatography and high-performance gel chromatography. The method of polymer detection for cefazolin sodium was established using the method of related substances HPLC as described in the Chinese Pharmacopoeia.

Streptomyces hygroscopicus 17997 produces the antiviral and antitumor ansamycin antibiotic, geldanamycin. Studies on geldanamycin biosynthetic pathway will provide good tools for genetic manipulation of the antibiotic-producing strain to improve the productivity or to facilitate making novel geldanamycin analogs. The structural similarities between geldanamycin and ansamycins such as rifamycin or ansatrienin suggest that both geldanamycin and ansamycins has a closely related pathways of biosynthesis and that biosynthetic system for geldanamycin is similar to the one of type I polyketide synthase (PKS) enzyme system. To explore the possible PKS genes involved in geldanamycin biosynthesis, the degenerate primers were designed according to the conserved sequence of KS-AT region from erythromycin and oleandomycin type I PKS genes. Cosmids containing multiple PKS genes (pCGBK2,4,6,10,11,18) were obtained by hybridization with the PCR products, which were amplified from S. hygroscopisus 17997 genomic DNA. The designed primers above were used for PCR. Development of a Streptomyces temperate phage phiC31-derivative KC515( tsrR) transduction system was carried out for identification of cosmids containing the PKS gene related to biosynthesis of geldanamycin. Several factors, mainly the Ca2+ and Mg + concentrations in different culture media affecting the frequency of gene transfection, were optimized .Transfection efficiency could reach up to 10(3) /microg DNA on YMG medium supplemented with 10mmol/L MgSO4. Reversely, the transfection efficiency decreased when YMG medium was supplemented with 30mmol/L MgSO4. Gene transfection system based on the integration-defective phage KC515 had been established for S. hygroscopicus17997. Recombinant phages (ph111, 258, 287, 116, 105) were constructed by insertion of the homologous to PKS gene fragments into the KC515 phage vector. Gene disruption experiments were performed by transduction of recombinant phages into S. hygroscopicus 17997 genome, and disruption of geldanamycin production was observed as a result of homologous recombination between the cloned insert in recombinant phage and the S. hygroscopicus 17997 genome by integration. Thiostrepton resistant transductants were selected and integration event was analyzed by Southern hybridization. The fermentation broth extracts from five resistant transductants were analyzed by TLC and HPLC. The results showed that only G16 mutant failed to produce geldanamycin. This result showed that the integration of the insert DNA fragment in recombinant phage phl6 into the chromosome of S. hygroscopicus disrupted the expression of the geldanamycin biosynthetic genes. The original cosmid pCGBK10 containing this cloned insert was predicted to encode PKS genes in the geldanamycin biosynthesis. This study laid the foundation for cloning the PKS genes involved in geldanamycin biosynthetic gene cluster from S. hygroscopicus 17997.
Alcohol Oxidoreductases ; genetics ; metabolism ; Bacterial Proteins ; genetics ; metabolism ; Bacteriophages ; genetics ; Benzoquinones ; metabolism ; Blotting, Southern ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Genetic Vectors ; genetics ; Lactams, Macrocyclic ; metabolism ; Multigene Family ; genetics ; physiology ; Polymerase Chain Reaction ; Streptomyces ; genetics ; metabolism

Objective To analyze the complications of coronary rotational atherectomy and to evaluate the safety of the procedure. Methods We evaluated the procedural and angiographic outcomes of 248 consecutive procedures to rotational atherectomy between January 2000 and October 2016. Results 27 cases(10.9％)were found to have rotational atherectomy related complications. Among these 27 cases,coronary spasm occurred in 2 cases(0.8％),no reflow in 8 cases(3.2％), coronary dissection in 5 cases(2.0％),burr entrapment in 6 cases(2.4％),wire breakage in 3 cases(1.2％), and coronary perforation in 3 cases(1.2％). There were no death,acute myocardial infarction and emergent coronary bypass graft.14 case(5.6％)had PCI-related myocardial infarction during hospital stay.In-hospital major adverse cardiaccerebral event(MACCE)rate was 5.6％. Conclusions Coronary rotational atherectomy can be performed with high success rates and procedure-related complications are rare.

OBJECTIVETo determine the expression of Cyclin B1, p34(cdc2) and the phosphorylation of survivin (p-survivin) in oral squamous cell carcinoma and oral submucosa fibrosis (OSF), and to discuss their possible role in carcinogenesis of OSF.

METHODSThe expression of Cyclin B1, p34(cdc2) and p-survivin were analyzed by Western blotting assay in 10 cases of normal oral mucosa epithelium, 40 cases of OSF epithelium and 42 cases of oral squamous cell carcinoma (OSCC) originated from OSF, respectively. Immunoprecipitation was used to confirm the relationship between the p34(cdc2) and survivin.

RESULTSThe expression of Cyclin B1, p34(cdc2), p-p34(cdc2) and p-survivin in OSF group were significantly higher than those in normal group (P < 0.05). The expression of these molecules showed significant different (P < 0.05) between the OSF and OSCC originated from OSF, but there was no significant difference among the early stage, the moderately advanced stage and the advanced stage of OSF. Immunoprecipitation assay confirmed the combination of p34(cdc2) and survivin.

CONCLUSIONSThe important molecules in G(2)/M phase-Cyclin B1, p34(cdc2) and p-survivin may play a key role during the mitosis and proliferation of OSF, which will be helpful in early diagnosis and therapy of carcinogenesis of OSF.

Adolescent ; Adult ; CDC2 Protein Kinase ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Case-Control Studies ; Cell Division ; Cyclin B1 ; metabolism ; Female ; G2 Phase ; Humans ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; metabolism ; Middle Aged ; Mouth Neoplasms ; metabolism ; pathology ; Neoplasm Staging ; Oral Submucous Fibrosis ; metabolism ; pathology ; Phosphorylation ; Young Adult

OBJECTIVETo study the value of detecting fetal congenital heart disease (CHD) using the five transverse planes technique of fetal echocardiography.

METHODSNine hundred and eighty-two high-risk pregnancies for fetal CHD were included in this study, the fetal heart was scanned with the five transverse planes technique of fetal echocardiography described by Yagel, autopsy was conducted when pregnancy was terminated. Blood from fetal heart was collected for fetal chromosome analysis. A close follow-up was given for normal fetal heart pregnancies and neonatal echocardiography was performed to check the accuracy of prenatal diagnosis.

RESULTS(1) Forty-six cases (4.68%) were found to have fetal heart abnormalities in this study, 69.56% of them were diagnosed by single four-chamber view, another 30.43% fetal CHD were found by combining other views; (2) Forty-one parents of prenatal fetuses with CHD chose to terminate pregnancy, thirty-two of them gave consent to conduct autopsy, 93.75% of which yielded unanimous conclusion between prenatal fetal echocardiography and autopsy; (3) Thirty-two of 46 cases underwent fetal chromosome analysis, 8 cases (25%) were found to have abnormal chromosome; (4) Five cases were found to have right ventricle and atrium a little bigger than those on the left side, with the unequal condition being the same after birth, but there were no clinical manifestations and they are healthy for the time being; (5) Nine hundred and thirty-six cases were not found with abnormality in this study, but one case was diagnosed with ventricular septal defect after birth, one case was diagnosed with patent ductus arteriosus, one case had atrial septal defect after birth.

CONCLUSIONS(1) The detected CHD rate was 4.68% by screening fetal heart with five transverse planes according to Yagel's description of high risk population basis for CHD. The coinciding rate of prenatal diagnosis and autopsy was 93.75%; (2) The sensitivity of detecting fetal heart abnormality is 92%, the specificity is 99.6% using the five transverse planes technique of fetal echocardiography; (3) Fetuses with mild or moderate disproportion of right and left side in the heart are potentially healthy babies.

Adult ; China ; Chromosome Aberrations ; Echocardiography ; methods ; statistics & numerical data ; Female ; Heart Defects, Congenital ; diagnostic imaging ; genetics ; Humans ; Infant, Newborn ; Pregnancy ; Sensitivity and Specificity ; Ultrasonography, Prenatal ; methods ; statistics & numerical data

OBJECTIVETo investigate the clinicopathologic manifestations of non-compaction of ventricular myocardium (NVM).

METHODSClinical data, electrocardiograms, echocardiography images and pathologic changes were studied in five cases of non-compaction of ventricular myocardium.

RESULTSThe patient's ages ranged from 29 to 57 years old, all were males. Abnormal electrocardiograms were obtained in all of the 5 cases. Among them, 3 were diagnosed using echocardiography. Histopathologic examination showed that there were abnormally coarse muscle trabeculation and deep recesses, interlacing in arrangement, over the inner wall of the heart chambers. The compacted myocardium became thinning down gradually from the base to the apex of the heart. The non-compacted myocardium bundles locating close to the endocardium were coarse and orderless in arrangement, nuclei were irregular and abnormal, nevertheless, the arrangement and appearance of the muscle bundles near by the pericardium part were essentially normal and the cell nuclei were evenly distributed.

CONCLUSIONThere are no specific clinical manifestations obtained in patients with non-compaction of ventricular myocardium, however, the pathologic changes are characteristic and a clinical diagnosis can be made by using echocardiography.

Adult ; Cardiomyopathy, Dilated ; diagnostic imaging ; pathology ; Cardiomyopathy, Hypertrophic ; diagnostic imaging ; pathology ; Diagnosis, Differential ; Electrocardiography ; Heart Ventricles ; pathology ; Humans ; Isolated Noncompaction of the Ventricular Myocardium ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Myocardium ; pathology ; Ultrasonography