Objective To investigate the value of real-time fluorescence detection technique of RNA (SAT) constantamplification in monitoring the effect of chemotherapy on patients with sputum positive pulmonary tuberculosis. Methods Sixty-two patients were selected, who were clinically diagnosed as the first-time retreatment for sputum smear-positive pulmonary tuberculosis and were hospitalized in our department from June 2015 to December 2016. After two-month standard anti-tuberculosis treatment, sputum samples were detected by Peng’s vessel acid-fast staining, BACTEC MGIT- 960 culture and strain identification, SAT detection. The BACTEC MGIT-960 culture and strain identification were used as gold standards, the value of SAT during the monitoring the therapeutic effect of the anti-tuberculosis drugs was assessed. Results After the treatment, 49 cases out of 62 showed positive results in mycobacterium tuberculosis culture test, among them 42 patients were diagnosed as human type mycobacterium tuberculosis, 7 patients were diagnosed as nontuberculous mycobacteria infection, and 13 cases showed negative results in mycobacterium tuberculosis culture. Thirty-two cases showed positive results in sputum Peng’s vessel acid-fast staining test, and 30 cases showed negative results. Forty-one cases showed positive results in SAT test and 21 cases showed negative results in SAT. SAT results were well concordant with sputum culture results (Kappa value=0.964), and the sensitivity, the specificity, the positive predictive value and the negative predictive value of SAT were 97.62%, 100%, 100% and 95.24% respectively. Peng’s vessel acid-fast staining results were badly concordant with MGIT-960 culture results (Kappa value=0.086), and the sensitivity, the specificity, the positive predictive value and the negative predictive value of Peng’s vessel acid-fast staining were 54.76%, 55.00%, 71.88% and 36.67% respectively. Conclusion SAT results can be used as good indices during the monitoring therapeutic effects of drugs used for the first-time retreatment in patients with sputum smear-positive pulmonary tuberculosis, which is worth promoting.

ObjectiveTo characterize the neural progenitor cell in the human amnion mesenchyme and epithelial layer with specific mark proteins of neural stem cell.MethodsExpressions of specific mark proteins of neural stem cell including nestin, glial fibrillary acidic protein (GFAP), musashi-1, vimentin and PSA-NCAM in human amnion tissue and cultured amniotic cells were determined by immunohistochemistry and immunofluorescence staining.ResultsExpressions of pluripotent neural stem cell specific makers (nestin, musashi-1, vimentin and PSA-NCAM) were detected in the human amnion mesenchyme and epithelial layer. In addition, cultured amniotic cells were expressed several neural stem cell specific markers including nestin, GFAP and PSA-NCAM. Nestin+ and GFAP+ double positive cells were identified in the human amnion tissue and cultured amniotic cells by immunohistochemistry and immunofluorescence staining.ConclusionSpecific mark proteins of neural stem cell are expressed in human amnion tissue and cultured amniotic cells.

cry1Ah1, one of holo-type cry genes, cloned in this laboratory from Bacillus thuringiensis strain has been patented in China, and it encoded a protein with strong insecticidal activity against certain lepidopteran insect pests, such as Chilo suppressalis. cry1Ah1 gene is exhibiting good application prospects. In order to improve the expression level of cry1Ah1 gene in rice, and investigate the effect of codon usage preference of gene expression, we designed five different optimized schemes for cry1Ah1 insecticidal critical fragment in accordance with bias of rice codon, to improve G+C content, removed the shear signal and unstable factors. Optimized cry1Ah1 genes were transformed into Escherichia coli Rosetta (DE3) respectively, and 65 kDa polypeptides was expressed normally in inclusion body separately. All of these expressed polypeptides showed insecticidal activity against 2nd-instar larvae of Plutella xylostella and neonate of Chilo suppressalis. After transformation with modified cry1Ah1 genes into Var nippobare, the transgenic rice seedlings were detected by PCR, the positive rate containing target gene was more than 87%. Afterwards, the results of real-time RT-PCR and ELISA assay indicated that the highest expression level of five modified cry1Ah1 genes was that using the highest frequent codons. Average expression amount of Cry1Ah1 polypeptides was 0.104% of total soluble proteins from the positive transgenic rice.
Animals ; Bacillus thuringiensis ; genetics ; metabolism ; Bacterial Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Codon ; genetics ; Endotoxins ; biosynthesis ; genetics ; Hemolysin Proteins ; biosynthesis ; genetics ; Insecticides ; Lepidoptera ; Oryza ; genetics ; Pest Control, Biological ; methods ; Plants, Genetically Modified ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

In past decade, hydrogen sulfide (H₂S) as a novel gasotransmitter, covered many fields in biological and medical research. However, there is no effective, convenient and high-throughput method for determination of circulatory H₂S until now. Here, we aim to develop an easy method for measurement of circulatory H₂S by modified methylene blue method. In the present study, we added Zn²⁺ to plasma sample to deposit H₂S, HS⁻ and S²⁻, as well as plasma protein, then used NaOH to re-dissolve plasma protein. ZnS deposition was re-dissolved by the addition of N, N-dimethyl-p-phenylenediamine, and the remnant protein was deposited by trichloroacetic acid. After centrifugation, ferriammonium sulfate was added to the supernatant fluid to generate methylene blue, which was analyzed by spectrophotometer at 665 nm. Using the present method, we found that most ions including sulfate and thiosulfate did not affect detection of H₂S concentration, but albumin (physiological concentration) reduced the detection value, which suggested the binding of serum albumin and a certain amount of H₂S. The relative recovery ratio of present method is 81.9%, which implies that the method is relative accurate for the determination of H₂S concentration in plasma or serum. H₂S levels of frozen plasma samples from 65 healthy volunteers detected by the present method were (13.93 ± 4.98) µmol/L. There was no obvious difference between the detection values of fresh and frozen samples from the same SD rats. These results suggest the modified methylene blue assay is stable, sensitive, convenient and high-throughput. The method can be used to analyze the circulatory H₂S in clinical and basic research.
Animals ; Blood Chemical Analysis ; methods ; Humans ; Hydrogen Sulfide ; blood ; Methylene Blue ; chemistry ; Phenylenediamines ; chemistry ; Rats ; Rats, Sprague-Dawley ; Sulfides ; chemistry ; Zinc Compounds ; chemistry

OBJECTIVETo compare the difference between the burn wound and diabetic ulcer wound, and to preliminarily analyze the nonhealing mechanism of diabetic unclear.

METHODSThe tissue of foot ulcer of diabete patients and skin wound tissues from burn patients were harvested. The levels of (FGF)2 and VEGF in the wound tissues were determined after tissue cultivation with enzyme-linked immunosorbent assay (ELISA). The changes in micro-vascular density (MVD) were examined by immunohistochemistry. Human umbilical vein endothelial cells were cultured in medium containing different components, and divided into following groups: A (with treatment of 5 mmol/L glucose for 7 days), B (with treatment of 30 mmol/L glucose for 7 days) and C (with treatment of 30 mmol/L Mannitol for 7 days) groups, then the level of VEGF protein was determined by ELISA.

RESULTSThe levels of FGF2 and VEGF protein in the burn wound were (59 +/- 3) ng/ml and (56 +/- 7) pg/ml, respectively, which were obviously lower than those in diabetic ulcer wound [(89 +/- 6) ng/ml, (108 +/- 5) pg/ml, P < 0.05]. There was also obvious difference in MVD between two kinds of wound (P < 0.05). The level of VEGF protein in both wounds were similar after the addition of FGF2 to the cell culture in vitro, while there were statistically significant difference 2 and 5 days after removal of FGF.

CONCLUSIONThe nonhealing mechanism of diabetic ulcer wound may be related to the inhibition of vacuolation and low expression of factors controlling vessel growth.

Burns ; complications ; metabolism ; pathology ; Cells, Cultured ; Diabetic Foot ; pathology ; Fibroblast Growth Factor 2 ; metabolism ; Foot Ulcer ; etiology ; pathology ; Humans ; Neovascularization, Physiologic ; Vascular Endothelial Growth Factor A ; metabolism ; Wound Healing

Objective To study the reliability of transcutaneous bilirubin (TCB) of different parts of neonates as predictive alarm of serum bilirubin ( SB ). Methods 132 cases of full-term neonates in Handan Central Hospital from May to July 2010 were divided into spontaneous delivery group and cesarean section group by random number. A KJ8000 transcutaneous bilirubinmeter was used to test their TCB at forehead, chest and abdomen on the fourth day after birth. The neonates were measured SB once TCB readings were more than 12.9 mg/dl. TCB of different parts and SB of the two groups were compared. Results The spontaneous delivery group had 17 cases of the. Neonates whose TCB readings were more than 12.9 mg/dl while the cesarean section group had 21 cases. TCB and SB of the same part by the same method showed no statistical significance between the two groups (t =0. 71, 2. 0, 1.25, 1. 0, 1.5;P ＞0. 05). TCB readings of chest were of no significant difference as compared with SB of the same group ( t =1. 72, 1. 33 ; P ＞ 0. 05 ). Conclusions SB of the spontaneous delivery group and the cesarean section group was of no significant difference. TCB reading of chest was closer to SB.

The purpose of this study was to investigate the vasorelaxing effect and mechanism of idoxifene (a new estrogen receptor modulator) on human internal mammary artery (HIMA). HIMA segments were harvested from men during coronary artery bypass grafting surgery. Patients with diabetes mellitus, hypercholesterolemia, hypertension, or smoking habit were excluded. The vasorelaxing effect of idoxifene on artery rings from HIMA with and without endothelium was measured by means of perfusion in vitro. Cumulative dose-response to idoxifene in the range of 0.01-10 micromol/L was observed in the presence and absence of NO synthase inhibitor L-NAME. It was also studied whether the vasodilation effect of idoxifene on HIMA was blocked by methylene blue (MB), an inhibitor of guanylate cyclase (GC). The results obtained from idoxifene were compared with those from 17beta-estradiol (E(2)). It was found that idoxifene caused a concentration-dependent relaxation on HIMA. The dose range was from 0.03 micromol/L (minimal vasodilatory concentration) to 3 mmol/L (maximal vasodilatory concentration). It was also found that the vasorelaxation effect of idoxifene on HIMA was dependent on endothelium. E(2) (0.1-100 micromol/L) also resulted in an endothelium-dependent vasorelaxation, but the vessels were 15-fold less sensitive to E(2) than to idoxifene in their vasorelaxation responses. The EC(50) for E(2) was 4.65+/-0.34 micromol/L, compared with 0.32+/-0.02 micromol/L for idoxifene. The mean maximal vasodilatory value of E(2) was 88.3+/-5.7%, compared with 88.6+/-7.2% for idoxifene. Pretreatment with L-NAME (100micromol/L) abolished idoxifene-induced vasodilation virtually by blocking nitric oxide production. The vasorelaxing effect of idoxifene disappeared in the presence of MB (10 micromol/L). These findings demonstrate that idoxifene results in an endothelium-dependent vasorelaxation of HIMA, like estrogen. The effect of idoxifene is more potent than that of traditional estrogen, and is possibly mediated by NO-GC-cGMP pathway.
Estrogen Antagonists ; pharmacology ; Humans ; Mammary Arteries ; drug effects ; physiology ; Tamoxifen ; analogs & derivatives ; pharmacology ; Vasodilation ; drug effects

Objective:To explore the expressions of zinc homeostasis-related proteins,G protein-coupled receptor 39(GPR39)and ANO1 mRNA in the sperm of patients with asthenozoospermia(AS),and analyze their correlation with sperm motility.Methods:We collected semen samples from 82 male subjects with PR+NP＜40％,PR＜32％and sperm concentration＞15 × 106/ml(the AS group,n=40)or PR+NP≥40％,PR≥32％and sperm concentration＞15 × 106/ml(the normal control group,n=42).We analyzed the routine semen parameters and measured the zinc content in the seminal plasma using the computer-assisted sperm analysis system,detected the expressions of zinc transporters(ZIP13,ZIP8 and ZNT10),metallothioneins(MT1G,MT1 and MTF),GPR39,and calcium-dependent chloride channel protein(ANO1)in the sperm by real-time quantitative PCR(RT qPCR),examined free zinc distribution in the sperm by laser confocal microscopy,and determined the expressions of GPR39 and MT1 proteins in the sperm by immunofluorescence staining,followed by Spearman rank correlation analysis of their correlation with semen parameters.Results:There was no statistically significant difference in the zinc concentration in the seminal plasma between the AS and normal control groups(P＞0.05).Compared with the controls,the AS patients showed a significantly reduced free zinc level(P＜0.05),relative expressions of MT1G,MTF,ZIP13,GPR39 and ANO1 mRNA(P＜0.05),and that of the GPR39 protein in the AS group(P＜0.05).No statistically significant differences were observed in the relative expression levels of ZIP8,ZNT10 and MT1 mRNA between the two groups(P＞0.05).The relative expression levels of GPR39,ANO1,MT1G and MTF mRNA were positively correlated with sperm motility and the percentage of progressively motile sperm(P＜0.05).Conclusion:The expressions of zinc homeostasis proteins(MT1G,MTF and ZIP13),GPR39 and ANO1 mRNA are downregulated in the sperm of asthenozoospermia pa-tients,and positively correlated with sperm motility.

BACKGROUNDBoth uniportal and triportal thoracoscopic lobectomy and sublobectomy are feasible for early-stage non-small cell lung cancer (NSCLC). The aim of this study was to compare the perioperative outcomes of uniportal and triportal thoracoscopic lobectomy and sublobectomy for early-stage NSCLC.

METHODSA total of 405 patients with lung lesions underwent thoracoscopic lobectomy or sublobectomy through a uniportal or triportal procedure in approximately 7-month period (From November 2014 to May 2015). A propensity-matched analysis, incorporating preoperative variables, was used to compare the short-term outcomes of patients who received uniportal or triportal thoracoscopic lobectomy and sublobectomy.

RESULTSFifty-eight patients underwent uniportal and 347 patients underwent triportal pulmonary resection. The conversion rate for uniportal and triportal procedure was 3.4% (2/58) and 2.3% (8/347), respectively. The complication rate for uniportal and triportal procedure was 10.3% and 9.5%, respectively. There was no perioperative death in either group. Most patients had early-stage NSCLC in both groups (uniportal: 45/47, 96%; triportal: 313/343, 91%). Propensity score-matching analysis demonstrated no significant differences in operation time, intraoperative blood loss, numbers of dissected lymph nodes, number of stations of lymph node dissected, duration of chest tube, and complication rate between uniportal and triportal group for early-stage NSCLC. However, the duration of postoperative hospitalization was longer in the uniportal group (6.83 ± 4.17 vs. 5.42 ± 1.86 d, P = 0.036) compared with the triportal group.

CONCLUSIONSUniportal thoracoscopic lobectomy and sublobectomy is safe and feasible, with comparable short-term outcomes with triportal thoracoscopic pulmonary resection. Uniportal lobectomy and sublobectomy lead to similar cure rate as triportal lobectomy and sublobectomy for early NSCLC.

Adult ; Aged ; Blood Loss, Surgical ; Carcinoma, Non-Small-Cell Lung ; pathology ; surgery ; Female ; Humans ; Length of Stay ; Lung ; pathology ; surgery ; Lung Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Operative Time ; Pneumonectomy ; adverse effects ; methods ; Prospective Studies ; Thoracic Surgery, Video-Assisted ; adverse effects ; methods ; Treatment Outcome

The present study was undertaken to investigate the role of glial glutamate transporter-1 (GLT-1) in the brain ischemic tolerance induced by cerebral ischemic preconditioning (CIP) by observing the effect of GLT-1 antisense oligodeoxynucleotides (AS-ODNs) on the neuro-protection of CIP against brain ischemic insult in rats. Wistar rats with permanently occluded bilateral vertebral arteries were randomly assigned to 7 groups: (1) Sham group: the bilateral common carotid arteries (BCCA) were separated, but without occluding the blood flow; (2) CIP group: the BCCA were clamped for 3 min; (3) Brain ischemic insult group: the BCCA were clamped for 8 min; (4) CIP+brain ischemic insult group: 3 min CIP was preformed 2 d prior to 8 min ischemic insult; (5) Double distilled water group: 5 muL double distilled water was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after the BCCA was separated (but without occluding the blood flow), respectively; (6) AS-ODNs group: 5 microL AS-ODNs solution was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after the BCCA was separated (but without occluding the blood flow), respectively. This group was further divided into 9 nmol and 18 nmol subgroups according to the doses of AS-ODNs; (7) AS-ODNs+CIP+brain ischemic insult group: 5 microL AS-ODNs solution was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after CIP, respectively. This group was also further divided into 9 nmol and 18 nmol subgroups according to the doses of AS-ODNs. The other treatments were the same as those in CIP+brain ischemic insult group. The effect of the AS-ODNs on the expression of GLT-1 was assayed by using Western blot analysis. The profile of delayed neuronal death (DND) of pyramidal neurons in the CA1 hippocampus was evaluated by using thionin staining under light microscope by determining the neuronal density (ND) and histological grade (HG). Western blot analysis showed that AS-ODNs injected into the lateral cerebroventricle inhibited the expression of GLT-1 in the CA1 hippocampus in a dose-dependent manner. Neuropathological evaluation showed that there was no apparent DND in sham and CIP groups. Obvious DND of pyramidal neurons was found in brain ischemic insult group, which was represented by an increase in HG and a decrease in ND. CIP effectively protected the pyramidal neurons in the CA1 hippocampus against DND normally induced by ischemic insult, which indicating that CIP induced ischemic tolerance on the pyramidal neurons in the CA1 hippocampus. However, the injection of AS-ODNs into the lateral cerebroventricle blocked the neuro-protection of CIP against DND induced by brain ischemic insult. These results further proved the role of GLT-1 in the brain ischemic tolerance induced by CIP in rats.
Animals ; Brain ; pathology ; Brain Ischemia ; drug therapy ; CA1 Region, Hippocampal ; pathology ; Excitatory Amino Acid Transporter 2 ; metabolism ; Ischemic Preconditioning ; Oligodeoxyribonucleotides ; pharmacology ; Oligonucleotides, Antisense ; pharmacology ; Pyramidal Cells ; metabolism ; Rats ; Rats, Wistar