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 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

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

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 observe the effects of acupotomy on cervicogenic headache and explore the function of cutaneous nerve entrapment in the mechanism of cervicogenic headache.

METHODSFrom October 2008 to June 2009, 82 patients with cervicogenic headache were treated with acupotomy. There were 23 males and 59 females, ranging in age from 17 to 73 years (averaged 41.57 years). The course of disease ranged from 0.5 to 50 years,with an average of 10.4 years. The location of treatment was occipitalia, both of left and right side: altogether were 8 points, including tenderness point of postmastoid; the mid-point between mastoid and C2 spinous process; the internal 1/3 attachment between occipital protuberance and mastoid process; the posterior midline of C2 spinous process open to 1.5-2 cm. The chief complaint of all patients was headache. PPI assessment rating was observed.

RESULTSThe total effective rate at 1 month after treatment was 81.70% (67/82). Recurrence of headache within 3 months after treatment was obvious. However, 17.07% (14/82) patients did not reoccur at 6 months after treatment.

CONCLUSION17.07% patients with cervicogenic headache recovered by acupotomy, so it shows cutaneous nerve entrapment plays an important role in the mechanism of cervicogenic headache. For the patients whose headache recurred at 3 months after treatment, increasing the treatment time and therapeutic range is suggested.

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Nerve Compression Syndromes ; therapy ; Occipital Bone ; surgery ; Post-Traumatic Headache ; therapy

Objective: To determine the molecular weight and p urity of porcine platelet-derived growth factor (pPDGF) and to investigate its effect on DNA synthesis of human umbilical vein endothelial cells. Metho ds: In the present experiment, the high performance liquid chromatograph y was used and the molecular weight and purity of pPDGF were studied. Human umbi lical vein endothelial cells was cultured and effects of pPDGF on DNA synthesis of endothelial cells was observed by 3H-TdR incorporation in vitro. Results: The findings of high performance liquid chromatography showed that the molecular weight of pPDGF was 29 120 and the purity was 89.46%, a nd pPDGF significantly promoted DNA synthesis of quiescent endothelial cells wit h a maximal response at a concentration of 40 ng/ml at 48 h. Conclusion: The molecular weight of pPDGF is 29 120, and it can promote DNA synthes is of cultured human umbilical vein endothelial cells.

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