10.3969/j.issn.1000-8179.2013.12.016

A themostable intracellular β-galactosidase from a thermophilic Bacillus stearothermophilus was purified by a combination of (NH4)2SO4 fractionation, ion-exchange (DEAE-22)and gel filtration (Sephades G-75). The optimum temperature and pH of the enzyme acivity were 60Cand pH6.4 respectively. The β-galatosidase activity exhibited thermosttability at 50 C. The enzyme was significaantly activated by alkali and alkali-earth metal ions. The activity was inhibited by Zn2+ 、 Fe3+ 、 Cu2+Reducing agents enhanced β- galactosidase activity. Thiol-binding agents drastically decreased the enzyme activity. The enzyme was specific for β-D glycosidic linkages,and the identity of the aglycone moiety also influenced enzyme activity. At 55Cthe Km for O-nitrophenyl-β-D-galactosidase (ONPG)and lactose were 2. 63mmol/L and 4.39mmol/L, respectively,and Vmax for both substrates were 1.93 × 10-5mmol. min-1 mg-1protein6.54 ×105 mmol. min-1. mg-1protein,respectively. The enzyme was inhibited by glucose (the products of lactose hydrolysis,ki 2.47mmol/L),but not by galactose. In addition,the enzyme possessed transgalactosylation activity. Galacto-oligosaccharides,both tri- and tetrasaccharide,were involved in the products during lactose hydrolysi

A themostable intracellular ? galactosidase from a thermophilic Bacillus stearothermophilus was purified by a combination of (NH 4) 2SO 4 fractionation,ion exchange (DEAE 22)and gel filtration (Sephades G 75).The optimum temperature and pH of the enzyme acivity were 60℃and pH6.4 respectively.The ? galatosidase activity exhibited thermosttability at 50 ℃.The enzyme was significaantly activated by alkali and alkali earth metal ions.The activity was inhibited by Zn 2+ 、 Fe 3+ 、 Cu 2+ Reducing agents enhanced ? galactosidase activity.Thiol binding agents drastically decreased the enzyme activity.The enzyme was specific for ? D glycosidic linkages,and the identity of the aglycone moiety also influenced enzyme activity.At 55℃the Km for O nitrophenyl ? D galactosidase(ONPG)and lactose were 2.63mmol/L and 4.39mmol/L, respectively,and Vmax for both substrates were 1.93?10 5 mmol.min 1 .mg 1 protein6.54?10 5 mmol.min 1 .mg 1 protein,respectively.The enzyme was inhibited by glucose (the products of lactose hydrolysis,ki 2.47mmol/L),but not by galactose.In addition,the enzyme possessed transgalactosylation activity.Galacto oligosaccharides,both tri and tetrasaccharide,were involved in the products during lactose hydrolysis.

Objective To establish and optimize AFLP reaction system used in providing necessary technique basis for genetic diversity analysis in Asarum sieboldii.Methods Leaves of A.sieboldii were used as experimental materials to analyze various essential elements of the whole process,such as quality of extracted DNA,time of restriction digest and ligation,concentration of Mg2+,primers and Taq DNA polymerase during PCR amplification etc.,so that the most optimal AFLP reaction system could be built up. Results The optimal AFLP reaction system of A.sieboldii has been constructed: in the genomic DNA extraction,mercaptoethanol being utilized and samples being incubated about 30 min at 65 ℃;the genomic DNA being digested by Trul Ⅰ and Pst Ⅰ for 3 h respectively;in PCR amplification,the final concentration of Mg2+ being 1.5 mmol/L and the volume of Taq DNA polymerase being 0.2 ?L.Conclusion The present reaction system for A.sieboldii is able to gain the favorable results of AFLP analysis and can be used in the genetic diversity research of the species.

Objective To investigate the protective effects of Erqi Decoction(EQD; mainly composed of Radix Aristolochiae Kaempferi, Radix Rhizoma Seu Flos Cypripedii, Cortex Fraxini, Cortex Phellodendri, Radix et Rhizoma Rhei) on the intestinal tract in rats with acute radiation intestinal injury and its mechanism. Methods Sixty SD rats were randomly divided into normal group, model group, EQD group and Baitouweng Decoction group (BD group), 15 rats in each group. The acute radiation enteritis model was established by exposing the whole abdomen to a total dose of 10 Gy of 6 MV higher-energy X-rays. EQD group and BD group were given intragastrical administration with corresponding medicine of EQD at the dose of 8.85 g·kg-1·d-1, BD at the dose of 4.69 g·kg-1·d-1 respectively, and the normal group and the model group were given intragastrical administration with the same volume of normal saline. The treatment lasted for 7 continuous days. After modeling, the morphological change of the proximal ileum tissue was observed under light microscope. Villus height, crypt depth, and thickness of the ileal mucosa and entire wall were measured by image analysis system. The myeloperoxidase (MPO) content in ileum tissue was determined by spectrophotometer, and the expression levels of caspase -3 and proliferating cell nuclear antigen (PCNA) in ileum tissue were determined by immunohistochemistry. Results EQD group and BD group had milder injuries of the ileal structure, and had higher villus height, crypt depth, and thickness of mucosa and entire wall than those in the model group (P <0.05), but there were no differences between the two medication groups(P > 0.05). MPO content in EQD group and BD group was decreased(P<0.05 compared with that in the model group), and MPO content in EQD group was lower than that in BD group. The expression levels of caspase-3 and PCNA were increased in EQD group and BD group(P < 0.05 compared with those in the model group), but there were no statistical differences between the two medication groups (P>0.05). Conclusion EQD has certain protective effects against radiation-induced intestinal damage, which mechanism is probably associated with relieving the local intestinal inflammatory reaction, accelerating intestinal epithelial cell proliferation, and inhibiting intestinal epithelial cell apoptosis.

Objective To compare the clinical effect of total laparoscopic and open liver resection for tumors in segments Ⅶ and Ⅷ.Methods The clinical data of patients with tumors in segments Ⅶ and Ⅷ of the liver who met the inclusion criteria and received operation at Affiliated Hospital of Nantong University from January 2011 to January 2015 were retrospectively analyzed.Among these patients, there were 17 cases who received total laparoscopic liver resection (LLR group), and 25 cases who received open liver resection (OLR group).Results LLR group has obvious advantages in aspects of the level of serum alanine transaminase (ALT) on 1st and 3rd day postoperation, the time anal exsufflation, the drainage volume of abdominal cavity in 3 days after operation and the postoperative hospital stay than those in OLR group (respectively t =-3.075,-3.175,-2.499,-2.088,-2.419, all P ＜ 0.05).There were no significant differences in blood transfusion rate, the resection margin to the tumor, the postoperative morbidity and the total medical cost between the two groups (x2 =1.437, t =-1.244, x2 =0.209, t =1.079, all P ＞ 0.05).Though the mean operative time and intraoperative blood loss of LLR group compared with OLR group increased obviously (respectively t =3.360, 2.189, all P ＜ 0.05).During the postoperative follow-up, there were no significant differences in the postoperative recurrence rate and the long-term survival rate in patients with malignant tumors (respectively x2 =0.240, 0.000, all P ＞ 0.05).Conclusion The therapeutic effect of total laparoscopic and open liver resection are equal in segments Ⅷ and Ⅷ hepatectomy, while, LLR has advantages of less trauma.

Objective To investigate influence of amlodipine /atorvastatin on atherosclerosis in the elderly with hypertension and hyperlipidemia.Methods Totally 108 cases with hypertension and hyperlipidemia in our hospital were randomly divided into control group and observation group (n =54 each).The control group received conventional antihypertensive treatment such as diuretics,angiotensin-converting enzyme inhibitors,calcium antagonists; the observation group received amlodipine and atorvastatin combination tablet as add-on therapy to the above drugs for 6 months.The blood lipid level and carotid artery ultrasound detection results were observed and compared between the two groups after treatment.Results The levels of TC[(4.23 ± 0.79)mmol/L vs.( 5.32 ±1.23)mmol/L],TG[(1.53±0.35) mmol/L vs.(5.32± 1.23) mmol/L],LDL-C [(3.19 ± 0.59)mmol/L vs.(4.07±0.79)mmo1/L]were decreased,while HDL-C [(1.53±0.95)mmol/L vs.(1.32±0.83)mmol/L]were increased in observation group after treatment as compared to control group (all P ＜0.05).After carotid artery ultrasonography,the LN[( 1.61± 0.76) scores vs.( 2.24 ± 0.89) scores ] and IMT [(0.76 ± 0.22)mm vs.(0.98 ± 0.28)mm] levels were lower in observation group after treatment than in control group (P＜0.05).Conclusions Amlodipine/atorvastatin combination can reduce blood lipids and alleviate atherosclerosis.It is suitable for elderly patients with hypertension and hyperlipidemia.

Objective:To analyze the clinical characteristics and risk factors of clopidogrel resistance after intracranial arterial stent implantation in patients with ischemic cerebrovascular disease.Methods:Retrospective case-control study was used.A total of 360 patients with ischemic cerebrovascular disease hospitalized in Beijing Tiantan Hospital from January 2017 to December 2018 were selected.All patients received intracranial arterial stenting and received double anti-platelet drugs after operation.Patients were divided into clopidogrel resistance group and non-resistance group according to the inhibition rate of platelet aggregation measured by thromboelastography.Clinical data and laboratory indicators of patients in the two groups were compared using two independent sample T tests and Mann-Whitney U tests, and possible risk factors of clopidogrel resistance were analyzed by Logistic regression.Results:White blood cell (WBC) count, neutrophils count, lymphocyte count, cholesterol, LDL cholesterol, and serum homocysteine levels in the clopidogrel non-resistant group were (6.58±1.45)×10 9/L, (4.01±1.05)×10 9/L, 1.83(1.49, 2.23)×10 9/L, (3.63±0.85) mmol/L, 1.93(1.53, 2.31) mmol/L, and 14.3(11.80, 17.00) μmol/L, respectively.Compared with the clopidogrel non-resistant group, the WBC count, neutrophils count, lymphocyte count, cholesterol, LDL cholesterol and serum homocysteine levels in the clopidogrel resistant group were all higher (7.19±1.53) ×10 9/L, (4.40±1.05) ×10 9/L, 2.03(1.63, 2.58)×10 9/L, (4.02±0.99) mmol/L, 2.04(1.68, 2.78) mmol/L and 15.90(12.25, 22.20) μmol/L, respectively.The difference was statistically significant ( t=3.277, t=2.867, Z=2.457, t=3.409, Z=2.705 and Z=2.220, respectively; P value were 0.001, 0.004, 0.014, 0.001, 0.007 and 0.026, respectively). Logistic regression analysis showed that high serum homocysteine ( OR=1.031, 95% CI 1.006-1.057, P=0.015) was an independent risk factor for clopidogrel resistance. Conclusion:Patients with clopidogrel resistance have high leucocyte, blood lipid, homocysteine and other clinical characteristics.High serum homocysteine is an independent risk factor for clopidogrel resistance.

Spectrophotometric seawater pH measurement system is an urgent need of device due to its quick speed and high precision. Based on spectrophotometric method and flow injection analysis technique, an automated system for pH measurement of seawater was developed by integrating pump and valve flow path, LED light source, flow cell and spectrophotometer. This measurement system effectively avoided bubbles. The indicator of concentration gradient in the seawater sample was used to correct for the pH perturbation caused by the indicator, thus the operation of system was simple and convenient. with this system, only 1. 5 min was needed for a sample measurement with a precision of 0. 0013 pH units and an offset of 0. 0059 pH units. This system could be used for the rapid determination of pH of seawater collected by laboratory or research ship with high precision pH values.