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.

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

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

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.

OBJECTIVETo explore the relationship of aging with the changes of endogenous carbon monoxide (CO), cGMP and cAMP contents in the penile tissues of rats.

METHODSTwenty-four male rats were equally divided into an 8-month, a 16-month and a 24-month group, and their penile erection was detected by injecting apomorphine, their penile cavernous body harvested, and the contents of CO, cAPM and cGMP detected by improved dual wavelength spectrophotometry.

RESULTSThe contents of CO, cAPM and cGMP were reduced with the increase of age, with statistically significant differences between the three age groups (P < 0.01).

CONCLUSIONAging significantly decreased the contents of CO, cAMP and cGMP in the penile tissues of the rats, which suggests that aging might play an important role in erectile dysfunction.

Aging ; physiology ; Animals ; Carbon Monoxide ; metabolism ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Male ; Penis ; metabolism ; Rats ; Rats, Wistar

Genotoxicity research takes an important place in traditional Chinese medicine safety evaluation. Genotoxicity test on traditional Chinese medicine has been paid great attention since 1970s. Currently, the most developed genotoxicity test methods included: bacterial reverse mutation test and mouse lymphoma assay which are used to detect relevant genetic changes, micronucleus test and chromosomal analysis which are used to measure chromosomal aberration, and single cell electrophoresis assay which is used to test DNA damage. This article reviews research progress on genotoxicity of traditional Chinese medicine, evaluation methods of genotoxicity, the problems and solutions on genotoxicity evaluation of traditional Chinese medicine, and new technique used in genotoxicity test.
Animals ; Biomedical Research ; Humans ; Medicine, Chinese Traditional ; adverse effects ; Mutagenicity Tests ; methods

Drug allergy and pseudoallergic reactions are main adverse drug reactions. Allergy is mainly induced by the immunogenicity of drug, drug metabolic products or drug additive. Pseudoallergic reactions may result from the irritation or activation of inflammatory material release. Pre-clinical evaluation of drug allergy and pseudoallergic reactions is included in immunotoxicity evaluation. Now there is no in vivo or in vitro method that could predict all kinds of allergy or pseudoallergic reactions due to the different mechanisms. In the past few years, FDA, SFDA OECD, ICH and WHO have published several guidelines on per-clinical immunotoxicity evaluation, however, no agreement has been reached on allergy and pseudoallergic reactions evaluation. This article reviews the requirements of allergy and pseudoallergic reactions in pre-clinical evaluation.
Drug Hypersensitivity ; diagnosis ; Humans ; Immune System ; drug effects ; Practice Guidelines as Topic