The recombinant plasmid pPIC-gpd-bgl-hyg was constructed, which contained GPD2 promotor and terminator from industrial yeasts Saccharomyces cerevisiae, ?-glucosidase gene (BGL1) from Sac-charomycopsis fibuligera and hyg from hygromycin as the selected marker. With the yeast’s high efficiency of homologous integrated, the BGL1 gene was successfully integrated into industrial yeasts S. cerevisiae. The recombined yeast could grow on the cultures with the cellobiose as a sole carbon source, and the ?-glucosidase activity achieved 0.764 U/mL after 48 hours’ cultivation. In the experiments of VHG ethanol fermentation, the cellobiose concentration in broth of recombined yeast was 80% lower than that of indus-trial yeast.

In gene manipulation, different selectable markers with various linkers are necessary. In order to get selectable markers directly, we constructed from pBlueScript SK(-) a series of particular plasmids, pSKsymKm, pSKsymBle, pSKsymEry, pSKsymHyg and pSKsymGm, each contains Kanamycin, Bleomycin, Erythromycin, Hygromycin or Gentamycin resistance cassette. By restriction enzyme digestion and gel extraction, any of five antibiotic resistance genes with specific ends can be conveniently obtained.

Objective:To systematically evaluate the efficacy and safety of neoadjuvant chemoradiotherapy (NCRT) plus surgery versus neoadjuvant chemotherapy (NCT) plus surgery in the treatment of advanced esophageal squamous cell carcinoma.Methods:Clinical controlled trials of comparing the treatment of NCRT plus surgery with NCT plus surgery for esophageal squamous cell carcinoma were electronically searched from the databases including PubMed, The Cochrane Library, EMbase, CBM, CNKI, WanFang and VIP from the inception of databases to January, 2019. Two reviewers independently screened the literatures, extracted data and assessed the risk of bias of the included studies. And then, a meta-analysis was performed by using RevMan 5.3 software.Results:A total of 8 clinical control studies were included, including 995 patients with esophageal squamous cell carcinoma. Meta-analysis results showed that compared with the NCT group, the R 0 resection rate was significantly higher ( OR=2.14, 95% CI: 1.03-4.45, P=0.040) and the pathological complete response (pCR) rate was significantly higher ( OR=4.19, 95% CI: 1.71-10.28, P=0.002) in the NCRT group. The incidence of postoperative complications ( OR=1.37, 95% CI: 0.76-2.48, P=0.300) and the risk of perioperative death ( OR=1.28, 95% CI: 0.58-2.83, P=0.54) were not significantly different between two groups. The long-term survival of patients with esophageal squamous cell carcinoma in the NCRT group was significantly better compared with that in the NCT group ( HR=0.77, 95% CI: 0.64-0.92, P=0.005). Conclusions:Compared with NCT plus surgery for advanced esophageal squamous cell carcinoma, NCRT plus surgery has higher R 0 resection rate and pCR rate, does not significantly increase the risk of perioperative complications or perioperative death, and significantly improves the long-term survival of esophageal squamous cell carcinoma patients.

The purification and the characteristics of an enzyme from Morganella morganii J-8, which could produce d-pseudoephedrine from 1-phenyl-2-methylamine-acetone, were performed in this study. In this research, first, cells were disrupted by ultrasonic treatment at 4 degrees C. The carbonyl enantioselective reductase was purified with a combination of ammonium precipitation, Phenyl Superose hydrophobic chromatography, DEAE anion exchange, and native polyacrylamide gel electrophoresis. The molecular mass of the purified enzyme subunit was estimated to be 42.5kD on sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE). The native molecular mass of the enzyme that was analyzed by high-performance liquid chromatography was found out to be 84.1 kD, which indicated that the enzyme was a dimmer. The purified enzyme was analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and the result showed that the purified enzyme had high homology with leucine dehydrogenase.
Bacterial Proteins ; chemistry ; isolation & purification ; metabolism ; Biocatalysis ; drug effects ; Chromatography, High Pressure Liquid ; Electrophoresis, Polyacrylamide Gel ; Enzyme Stability ; drug effects ; Hydrogen-Ion Concentration ; Kinetics ; Leucine Dehydrogenase ; metabolism ; Metals, Heavy ; pharmacology ; Molecular Weight ; Morganella morganii ; enzymology ; metabolism ; Oxidoreductases ; chemistry ; isolation & purification ; metabolism ; Pseudoephedrine ; chemistry ; metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Stereoisomerism ; Temperature

OBJECTIVETo evaluate therapeutic effect of the needle-knife closed solution combined with minor adjusting of spine for treatment of neck-shoulder syndrome.

METHODSFrom April 2010 to August 2011,120 patients with neck-shoulder syndrome were treated with the needle-knife closed solution combined with minor adjusting of spine, and included 45 males and 75 females and aged from 40 to 68 years old. The disease course was from 3 days to 10 years. After the operation, all patients taken the medicine of activating blood circulation herbs. At the 3rd, 7th, 10th day after operation, spinal rotation massage was performed on these patients. After the healing of the needle points, traditional Chinese medicine herb fumigation was applied on the needle points, and the patients were directed to do the cervical spine exercise. Therapeutic effect of the patients was evaluated by the neck disability index (NDI).

RESULTSAll patients were followed up after 3 weeks' treatment. The pain of neck-shoulder was relieved, and the range of motion was improved,the NDI score lowered from 49.30 +/- 1.35 before treatment to 10.15 +/- 1.18 at 3 weeks after treatment (t = 2.116, P < 0.05).

CONCLUSIONThe needle-knife closed solution combined with minor adjusting of spine for the treatment of neck-shoulder syndrome can relieve the pain in the neck-shoulder and improved the motion of the neck. The key for the effect is accurate location before operation, sufficient adhesion solution during the operation and spinal minor adjusting after operation.

Adult ; Aged ; Female ; Humans ; Male ; Manipulation, Spinal ; methods ; Middle Aged ; Myofascial Pain Syndromes ; therapy ; Neck Pain ; therapy ; Shoulder Pain ; therapy

OBJECTIVETo investigate the effects of CO release inhibitor zinc protoporphyria IX (ZnPP IX) and NO release inhibitor L-NAME on the content of cGMP in the penile tissue of rats.

METHODSThirty Wistar rats were randomly divided into a normal control, a ZnPP IX, and an L-NAME group, given saline (1 ml/kg/d), ZnPP IX (45 micromol/kg/d) and L-NAME (50 mg/kg/d), respectively, for 7 days. Then all the rats were killed, homogenate made from their penile tissues and detected for the contents of NOS, NO, CO and cGMP.

RESULTSThe contents of CO, NOS, NO and cGMP were all reduced in both the ZnPP IX and L-NAME groups as compared with the control group (P < 0.05).

CONCLUSIONZnPP IX and L-NAME can reduce the concentrations of CO and NO in the penile tissues of rats, and consequently the content of cGMP.

Animals ; Carbon Monoxide ; antagonists & inhibitors ; metabolism ; Cyclic GMP ; metabolism ; Male ; NG-Nitroarginine Methyl Ester ; pharmacology ; Nitric Oxide ; antagonists & inhibitors ; metabolism ; Penis ; drug effects ; metabolism ; Protoporphyrins ; pharmacology ; Rats ; Rats, Wistar

Objective:To evaluate therapeutic effect of the needle knife closed solution combined with minor adjusting of spine for treatment of neck shoulder syndrome. Methods:From April 2010 to August 2011,120 patients with neck shoulder syndrome were treated with the needle knife closed solution combined with minor adjusting of spine ,and included 45 males and 75 females and aged from 40 to 68 years old. The disease course was from 3 days to 10 years. After the operation ,all pa-tients taken the medicine of activating blood circulation herbs. At the 3rd,7th,10th day after operation,spinal rotation massage was performed on these patients. After the healing of the needle points ,traditional Chinese medicine herb fumigation was ap-plied on the needle points,and the patients were directed to do the cervical spine exercise. Therapeutic effect of the patients was evaluated by the neck disability index (NDI). Results:All patients were followed up after 3 weeks'treatment. The pain of neck shoulder was relieved,and the range of motion was improved,the NDI score lowered from 49.30±1.35 before treatment to 10.15±1.18 at 3 weeks after treatment (t=2.116,P<0.05). Conclusion:The needle knife closed solution combined with minor adjusting of spine for the treatment of neck shoulder syndrome can relieve the pain in the neck shoulder and improved the mo-tion of the neck. The key for the effect is accurate location before operation ,sufficient adhesion solution during the operation and spinal minor adjusting after operation.

OBJECTIVEA molecular technique based on quasispecies analysis for tracing postexposure HIV transmission was applied in an investigation of a possible case of HIV transmission after blood transfusion.

METHODSSixteen plasma specimens were collected from 3 HIV infections (T1-T3) involved in a possible HIV transmission chain and 13 HIV/AIDS (C1-C13) controls. The RNAs were extracted and then amplified by RT-PCR, the PCR products were cloned and sequenced.BioEdit 6.0.7 and MEGA 4.0 software were used to analyze gene sequences, calculate gene dispersion ratio and construct phylogenetic tree.

RESULTSThe sequences of 13 specimens were successfully obtained.The HIV strains from T1, T2 and T3 were CRF07_BC recombinants, those from 5 out of the 6 controls lived in the same city with T2 and T3 were CRF07_BC recombinants as well, while those from 4 controls living in the same city with T1 were CRF01_AE recombinants. Compared with the clone sequences from T1, the mean gene dispersion ratio of T2 was the least (2.0%), followed by C12 (2.8%) , T3 (2.9%) and others. The phylogenetic tree showed that all clones from T1, T2, T3 and C12 might cluster together,and implied that the direction of HIV transmission was from T3 to T2, and then to T1.

CONCLUSIONThe results support the possible epidemiological clue that HIV was transmitted from T3 to T2, and then to T1, indicating that molecular epidemiological investigation could provide more direct evidence for tracing postexposure HIV transmission.

Female ; HIV ; genetics ; HIV Infections ; epidemiology ; genetics ; transmission ; Humans ; Male ; Molecular Epidemiology ; Phylogeny ; RNA, Viral ; genetics ; Sequence Analysis, RNA ; Transfusion Reaction

Objective To study clinical data of patients with acute myocardial infarction (AMI) induced by left main (LM) artery occlusion,and to find out the clinical characteristics.Method From Janurany 1995 to May 2006,there were 15 patients with AMI related with LM obstruction from 1793 AMI patients,in whom primary PCls were performed.All patients were male with a mean age of (60.6?10.4) years old (ranging from 43 to 85 years old).The clinical and interventional data were retrospectively analyzed.Results Cardiogenic shock occurred in 10 patients at admission.Primary PCls were performed with intraaortic balloon pump (IABP) in all patients.Nine patients died during short follow-up,and the mortality was 60%.Three-month follow-up was made in the all survival patients,and one patient died after 4 years.Analysis showed good collateral circulation flow from right coronary artery to left coronary artery before operation affected the short-term prognosis.Conclusions AMI related with left main artery obstruction was critically fatal with high occurrence of cardiogenic shock and high mortality.Prognosis differs at,cording to different degree of collateral circulation.Primary PCI with IABP provides a promising strategy to improve clinical outcomes.

Pseudoallergic reactions of Qingkailing injection (QKLI) was assessed by vascular hyperpermeability which were indicated by ear blue staining in ICR mice after single intravenous injection of QKLI mixed with Evans blue (EB) and skin blue spot formation in SD rats after intradermal injection of QKLI and intravenous injection of EB. In addition, QKLI-induced histamine, VEGF, TNF-alpha release was measured after ICR mice received the single dosing of QKLI iv. The mild vascular hyperpermeability characterized by ear blue staining could be observed in mice after intravenous injection of QKLI and EB. Intracutaneous injection of 50 micro L of test solution containing QKLI (25,50 microL) in rat back skin caused obvious local vascular hyperpermeability at the injection sites so as to result the larger diameters of blue spots than that in negative control group (P <0. 01). QKLI induced a significant increase of VEGF and a slight elevation of histamine in mice after intravenous administration, while TNF-alpha showed no change after QKLI iv. The results in this study indicated that both intravenous injection and intracutanous injection of QKLI could induce vascular hyperpemeability so as to cause pseudoallergic reaction in mice and rats. QKLI-induced pseudoallergic reaction may be associated with the release of histamine and VEGF.
Animals ; Drug Hypersensitivity ; blood ; etiology ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Histamine ; blood ; Injections ; Male ; Mice ; Rats ; Skin ; drug effects ; immunology ; Tumor Necrosis Factor-alpha ; blood ; Vascular Endothelial Growth Factor A ; blood