We constructed plasmid pMTac to overexpress 3-ketosteroid-Δ1-dehydrogenase (KSDD) in Mycobacterium neoaurum JC-12 for improving androst-1,4-diene-3,17-dione (ADD) production. To construct pMTac, pACE promoter on pMF41 was replaced by tac promoter, and then four recombinants were constructed, which were M. neoaurum JC-12/pMF41-gfp, M. neoaurum JC-12/pMTac-gfp, M. neoaurum JC-12/pMF41-ksdd and M. neoaurum JC-12/pMTac-ksdd. Fluorescence detection results show that much more green fluorescent protein (GFP) was expressed in M. neoaurum JC-12/pMTac-ksdd than M. neoaurum JC-12/pMF41-ksdd. The activity of KSDD was 2.41 U/mg in M. neoaurum JC-12/pMTac-ksdd, 6.53-fold as that of M. neoaurum JC-12 and 4.36-fold as that of M. neoaurum JC-12/pMF41-ksdd. In shake flask fermentation, ADD production of M. neoaurum JC-12/pMTac-ksdd was 5.94 g/L, increased about 22.2% compared to the original strain M. neoaurum JC-12 and 12.7% to M. neoaurum JC-12/pMF41-ksdd. AD (4-androstene-3,17-dione) production of JC-12/pMTac-ksdd was 0.17 g/L, decreased 81.5% compared to M. neoaurum JC-12 and 71.2% to M neoaurum JC-12/pMF41-ksdd. In the 5 L fermenter, 20 g/L phytosterols was used as substrate, ADD production of M. neoaurum JC-12/pMTac-ksdd was improved to 10.28 g/L. pMTac is favorable for expressing KSDD in M. neoaurum JC-12, and overexpression of KSDD has beneficial effect on ADD producing, and it is the highest level ever reported using fermentation method in M. neoaurum.
Androstadienes ; metabolism ; Fermentation ; Industrial Microbiology ; Mycobacterium ; Oxidoreductases ; genetics ; metabolism ; Phytosterols ; metabolism ; Plasmids

Objective To study the clinical significance of the serum angiopoietin-2(Ang-2) in the diagnosis,recurrence,invasion and metastasis of gastric cancer.Methods The serum Ang-2 and carcino-embryonic antigen (CEA) levels in 158 patients with gastric cancer (gastric cancer group) and 30 normal controls(control group) were measured by enzyme linked immunosorbent assay(ELISA) technique respectively.The serum Ang-2 and CEA levels were also measured 2 weeks after operation in gastric cancer group and reexamined in the recurred gastric cancer patients in 2 years after operation (recurred and metastasis group).The correlation between the serum Ang-2 level and pathologic c haracterization of gastric cancer was evaluated.Results The serum Ang-2 and CEA levels in gastric cancer group [ (331.8 ±64.3),(42.6 ±37.3)μg/L] and recurred and metastasis group [(318.7 ±72.9),(40.5 ±36.7)μg/L] were significantly higher than those in control group [ (187.4 ± 32.7),(4.2 ± 3.1 )μ g/L] (P ＜ 0.01 ),and the serum Ang-2 level 2 weeks after operation [ (211.6 ± 75.1 ) μ g/L ] was significantly decreased to the control group (P ＞ 0.05 ),while the serum CEA level [ (33.4 ± 30.6) μ g/L ] was still significantly higher than the control group (P ＜ 0.01 ).The sensitivity of the serum Ang-2 for diagnosis of gastric cancer was markedly higher than that of the serum CEA (P ＜ 0.01 ).There was correlation between serum Ang-2 and degree of tumor differentiation,TNM pathological staging,lymphatic metastasis,invasion depth and tumor size (p ＜0.01 ),but there was no correlation between serum Ang-2 and tissue classification and location of gastric cancer (P＞ 0.05).Conclusion The serum Ang-2 level is suggested to be a valuable gastric cancer marker and conduce to the diagnosis of gastric cancer,the monitoring of recurrence after operation and evaluation of prognosis.

Objective To determine if Shenfu injectio(SFI)has any analeptic action in patients emerging from general anesthesia.Methods Eighty-six ASAⅠorⅡadult patients undergoing elective abdominal surgery under general anesthesia were randomly divided into 2 groups(n=43 each):SFI group and control group.The patients were premedicated with intramuscular phenobarbital 0.1g and atropine 0.5mg.Anesthesia was induced with propofol 2 mg?kg~(-1),fentanyl 4-5?g?kg~(-1) and vecuronium 0.1 mg?kg~(-1) and maintained with propofol infusion(2-4 mg?kg~(-1)?h~(-1)),0.5%-1.0% isoflurane inhalation and intermittent i.v.boluses of fentanyl and vecuronium.The patients were intubated and mechanically ventilated.The propofol infusion and isoflurane inhalation were stopped during skin closure.The patients were still unconscious and on mechanical ventilation at the end of surgery and transferred to PACU with a tube in trachea.As soon as the patients reached the PACU,SFI 1 ml?kg~(-1) in Ringer's solution 100 ml was infused over 10 min.In control group the patients received Ringer's solution 100 ml without SFI.The following times were recorded:(1)the time when the patients opened their eyes on command;(2)the time when mechanical ventilation was stopped;(3)the time when oxygen inhalation was stopped;(4)the extubation time;(5)the time of staying in PACU.Venous blood samples were taken before(T_0) and 5,15 and 45 min(T_(1,2,3))after SFI infusion for determination of plasma?-endorphin concentration.Results The awakening time,the mechanical ventilation time,oxygen inhalation time,extubation time and duration of PACU stay were significantly shorter in SFI group than in control group.There were no significant differences in MAP and HR after SFI between the two groups.The plasma?-endorphin concentration was significantly higher in group SFI than in control group.Conclusion Shenfu injectio can make patients emerging from general anesthesia faster.

Effects of Cr 6+ concentration and culture time on four heavy metal removing strains,stability of these four strains removing Cr 6+,configuration changes inside or outside their cells,effects of Cr 6+ on soluble reductive sugar inside their cells,and effect of several factors on these strains had been studied,and the Cr 6+ resistance mechanisms of these strains have been discussed elementarily. The results showed that the Lethality of these strains caused by Cr 6+ was similar with one another, namely, increasing at first, then decreasing, and finally increasing again as culture time passed. Acclimatization of Candida sp. was better than Sporobolomycetaceae sp.,and the Cr 6+ resistance of Sporobolomycetaceae sp. 7-3 was the best of the four. The research also demonstrated that the metabolic activity of these strains had been influenced by Cr 6+ in a certain extent. Scanning electron microscopy,transmission electron microscopy,and atomic force microscopy observations approved that removal of Cr 6+ by Candida sp. was depended on both surface adsorption and intracellular accumulation. Effects of Cr 6+ concentration, pH, culture time, nitrogen source, carbon source and adsorption time on these strains are not the same.

Objective:To search for a method for radical resection of pancreatic and duodenal malignancy involving the mesentery root and for the long post-operation survival of patients.Methods:From Jan.2004 to Aug.2006,a total of 26(16 male and 10 female. aged 27-70)patients with pancreatic and duodenal malignancy involving the mesentery root were treated in our department.The patients included 3 with duodenal malignancy and 23 with pancreatic malignancy.Curative resection was performed by the extended pancreaticoduodenetomy(Whipple procedure)combined with mesentery root resection(MRR)for all patients.The outcomes,safety and the post-operation survival rate were analyzed retrospectively.Results:Thirteen patients were treated with Whipple procedures combined with MRR,9 were treated with partial portal vein/superior mesenteric vein(PV/SMV)and reconstruction of the vessel,and 4 patients received pre-shunt between PV and SMV with artificial vessel graft before the extended Whipple and MRR procedures.The operation time was 2.5 to 7(4.4?1.1)hour,and blood loss was 300 to 5 000(1892?1414)ml with the blood transfusion of 0 to 5 600(2 100?1 586)ml.There was no death in our group and 7(27%)had post-operation complication.The post-operation hospital stay was 10 to 30 days.The pathologic examination showed negative surgical margins for all specimens.The tumor size was 4 to 10 (6.17?2.03)cm.After a follow-up of 9 to 38 months,the pain was relieved in all patients.One of the 3 patients with duodenal adenocarcinoma had liver metastasis at 10 months after operation,and the other 2 survived 10 months and 27 months without evidence of tumor reccurence.The patient with pancreatic micro-adenocarcinoma died of local reccurence 9 months after operation.The patient with neuroendocrine carcinoma died of organ failure 24 months after operation.The patient with lymphoma have survived for 24 months after operation.The 1-year and 2-year accumulated survival rates in the 20 cases with pancreatic ductal cancer were 86.6% and 45.6%. respectively.Conclusion:The extended Whipple procedure with MRR is safe and effective.It can obtain R0 resection in patients with malignant tumors(over 5 cm in diameter)in the head,neck and uncinate process of the pancreas and duodenal.

Objective To detect the Yersinia pestis plasmid and molecular weight along the Qinghai-Tibet Railway.Methods Yersinia pestis plasmids molecular weight detected and analyzed using alkaline lysis,phenol-chloroform extraction of Yersinia pestis plasmid by agarose gel electrophoresis.Results The 18 Yersinia pestis strains of Qinghai-Tibet Railway contained 6×106,45×106,52×106,65×106,92×106plasmid,varing in the range of the 52×106-92×106.Conclusions The Yersinia pestis of Qinghai-Tibet Railway has a standardplasmid graphics,with the biggest Yersinia pestis plasmid changing in a certain regular degree,which providessignificance in the study of plague natural foci of the spatial structure and the genetic.characteristics of Yersiniapestis.

OBJECTIVETo analyze the clinical factors that contribute to lowered bone mineral density (BMD) of the lumbar vertebrae in type 2 diabetic patients.

METHODSForty type 2 diabetic patients with osteoporosis and 40 age- and gender-matched healthy individuals, selected using propensity score method, were examined for BMD of the L1 to L4 vertebrae and the left femur. Age, gender, course of the disease, body mass index (BMI), fasting blood glucose, serum calcium, serum phosphate, urinary calcium, and urinary phosphate were compared between the two groups.

RESULTSBMD of the L1 to L4 vertebras was significantly lower in the diabetic group than in the healthy individuals (P<0.05). In the diabetic patients, BMD showed an obvious difference between male and female patients and was negatively correlated with the course of the disease but positively with BMI, fasting blood glucose, urinary calcium and urinary phosphate; BMD was not correlated with age, serum calcium or serum phosphate in these patients.

CONCLUSIONOsteoporosis in type 2 diabetic patients is closely related with gender, BMI, course of the disease and poorly controlled glucose level.

Aged ; Arthroplasty, Replacement, Knee ; methods ; Female ; Femur ; surgery ; Humans ; Osteoarthritis ; surgery ; Osteotomy ; methods

OBJECTIVETo investigate the chemical constituents of the extract of Galium verum.

METHODThe compounds were isolated by chromatography and identified by spectral data.

RESULTThe eleven compounds obtained were identified as (+)-pinoresinol 4,4'-O-bis-beta-D-glucopyranoside (1), epipinoresinol (2), (+) -medioresinol (3), isorhamnetin (4), isorhamnetin 3-O-alpha-L-rhamnopyranosyl-(1-6)-beta-D-glucopyranoside (5), diosmetin (6), diosmetin 7-O-beta-D-glucopyranoside (7), quercetin-3-O-beta-D-glucopyranoside (8), ursolic acid (9), ursolic aldehyde (10) and rubifolic acid (11).

CONCLUSIONCompounds 1-5 and 9-11 were isolated from this genus for the first time.

Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Galium ; chemistry

OBJECTIVETo observe the clinical efficacy difference between segmentation scraping and conventional acupuncture for cervical spondylosis (CS) so as to provide effective treatment method.

METHODSEighty-five cases of cervical type of CS were randomly divided into a scraping group (44 cases) and an acupuncture group (41 cases). The segmentation scraping therapy was used in the scraping group. The scraping group was treated with focusing on scraping the head and joint part of neck and occiput in the upper cervical spine injury, and focusing on scraping the lower section of cervical and shoulder in the lower cervical spine injury, once every seven days, totally for 3 times. In the acupuncture group, Fengchi (GB 20),Wangu (TE 5), Tianzhu (BL 10),Neck-Jiaji (EX-B 2), etc. were selected,once daily,for 15 days. The visual analogue scale (VAS) was used to evaluate the immediate analgesic effect after the first treatment and the clinical efficacy was observed after the end of treatment.

RESULTSAfter the first treatment, the score of VAS was decreased significantly in the scaping group (P < 0.01), but there was no significant difference in the acupuncture group compared with those before treatment (P > 0.05); the score of VAS in the scaping group after the first treatment was lower than that in the acupuncture group (3.66 +/- 0.74 vs 5.43 +/- 0.35, P < 0.01). Compared with before treatment, the scores of VAS were decreased significantly after treatment in two groups (both P < 0.01), but without significant difference between two groups (P > 0.05); the effective rate was 95.5% (42/44) in the scaping group and 87.8% (36/41) in the acupuncture group, the curative effects were similar (P > 0.05).

CONCLUSIONBoth of scraping and acupuncture therapies have good analgesic effect for cervical spondylosis, and overall effects are similar, but the immediate analgesic effect of scraping thrapy is better than that of conventional acupuncture.

Acupuncture Points ; Acupuncture Therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Neck Pain ; therapy ; Spondylosis ; therapy ; Treatment Outcome