Genes related to trehalose biosynthesis from a bacterial strain Micrococcus luteus which can convert partially hydrolyzed starch into trehalose were cloned.Full sequence of gene (MtreY) encoding trehalose maltooligosyl trehalose synthase (MTSase) and partial sequence of gene (MtreZ) encoding maltooligosyl trehalose trehalohydrolase (MTHase) were got using PCR combined non-random shotgun method.Sequence analysis of MtreY predicts a 2370bp open reading frame encoding a protein of 790 amino acids with a predicted molecular weight of 86734 Da.Homologous analysis shows that this new gene has the same conservative motifs with ?-amylase family enzymes.The MtreY gene was expressed in E.coli, and the expression product has the anticipative enzyme activity.

OBJECTIVETo study the chemical constituents from the stems and leaves of Calophyllum inophyllum.

METHODThe compounds were isolated by column chromatography on silica gel, Sephadex LH-20 and preparative TLC. Their structures were elucidated by chemical methods and NMR, MS spectroscopic data.

RESULTNine compounds were identified as 2-hydroxyxanthone (1), 4-hydroxyxanthone (2), 1, 5-dihydroxyxanthone (3), 1, 7-dihydroxyxanthone (4), 1, 3, 5-trihydroxy-2-methoxyxanthone (5), 6-deoxyjacareubin (6), amentoflavone (7), kaempferol-3-O-alpha-L-rhamnoside (8) and quercetin-3-O-alpha-L-rhamnoside (9).

CONCLUSIONCompounds 8 and 9 were isolated from the genus Calophyllum and compounds 1, 2, 4-6 were isolated from this plant for the first time.

Calophyllum ; chemistry ; Chromatography ; methods ; Flavonoids ; analysis ; chemistry ; isolation & purification ; Glycosides ; analysis ; chemistry ; isolation & purification ; Kaempferols ; analysis ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; methods ; Mass Spectrometry ; methods ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Pyrans ; analysis ; chemistry ; isolation & purification ; Quercetin ; analogs & derivatives ; analysis ; chemistry ; isolation & purification ; Xanthenes ; analysis ; chemistry ; isolation & purification

OBJECTIVETo assess the effect of the autologous venous external stents on intimal hyperplasia of the vein grafts in rabbits.

METHODSThirty-six male New Zealand white rabbits, aged 5 months and weighing 2.8 to 3.0 kg, were randomly divided into 3 groups: group A, group B and group C, with 12 rabbits in each group. First, a section about 6 cm long of vein was cut from the right external jugular vein of each rabbit and severed to have 3 equal-length segments. Next, each distal segment prepared for anastomosis. The proximal segment invaginating middle segment in group A and only middle segment in group B were used for the external stent. Later, the left common carotid artery was separated from surrounding tissue, from it a section about 0.5 cm long was cut away. Finally, the vein graft was inverted and end-to-end anastomosed to the two ends of the artery with a 9-0 suture. After bloodstream re-established, the diameter of each vein graft was measured. At 2 and 4 weeks postoperative, the graft veins were cut off and histologically examined by the means of HE staining and Masson staining. The smooth muscle cells (SMC) proliferation was studied by the immunohistochemical detection of proliferating cell nuclear antigen.

RESULTSAfter bloodstream re-established, the diameters of vein graft of group A and group B and group C were (1.6 +/- 0.3) mm, (2.2 +/- 0.4) mm and (2.6 +/- 0.6) mm respectively (P < 0.05). At 4 weeks postoperative, the data of the ratio of intima to media thickness and the index of the proliferating cells of the intima were as follow: group A (1.01 +/- 0.07 and 6.84 +/- 1.98), group B (1.32 +/- 0.08 and 11.01 +/- 2.61), group C (1.55 +/- 0.03 and 14.96 +/- 4.14). Both the data of group A were obviously less than that in group B, and that of group B was less than group C (P < 0.05).

CONCLUSIONThe autologous venous two-layer external stents inhibit intimal hyperplasia of the vein grafts.

Animals ; Hyperplasia ; pathology ; prevention & control ; Male ; Rabbits ; Stents ; Transplantation, Autologous ; Tunica Intima ; pathology ; Veins ; pathology ; transplantation

OBJECTIVETo study the proportion and mechanism of relieving asthma of drug partnership comprising herbal Ephedrae & Pheretima.

METHODTo study relaxant effect on 10 micromol x L(-1) carbachol (CCh) and 10 micromol x L(-1) histamine (His) precontracted isolated tracheal rings and lowering effect on short-circuit current (Isc) increase induced by 10 micromol x L(-1) CCh with 3 proportions of 1:1, 1:3, 1:9 extract.

RESULT1:3 proportions dose-dependently relaxed CCh-precontracted isolated tracheal rings, IC50 of 1:1, 1:3 is 7.5, 15 mg x mL(-1) respectively, 1:9 could not produce 50% inhibition effect on CCh-evoked contraction; 3 proportions also dose-dependently relaxed His-precontracted isolated tracheal rings, IC50 of 1:9, 1:3 and 1:1 is 0.19, 0.61, 1.8 mg x mL(-1) respectively. On the other hand,the orders potency of the decrease effect on CCh-evoked short circuit current increase is 1:3 > 1:1 > 1:9. The difference is not significant (P < 0.05).

CONCLUSIONHerbal Ephedrae & Pheretima had tracheal muscle relaxant and epithelium ion secretion inhibition effect, its mechanism of relieving asthma involved anti-CCh and anti-His effect 1:3 was the most appropriate dosage ratio in the anti-asthmatic drug partnership.

Animals ; Anti-Asthmatic Agents ; administration & dosage ; pharmacology ; Asthma ; physiopathology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Ephedra sinica ; chemistry ; Guinea Pigs ; Histamine Antagonists ; pharmacology ; In Vitro Techniques ; Male ; Materia Medica ; administration & dosage ; isolation & purification ; pharmacology ; Muscle Relaxation ; drug effects ; Muscle, Smooth ; drug effects ; Oligochaeta ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the value of application of Bioflex dynamic stabilization system in treating multi-segment lumbar degenerative disease.

METHODSClinical datas of 13 patients with multi-segment lumbar degenerative disease (8 males and 5 females,ranging in age from 51 to 72 year with an average of 65.0) were retrospectively analyzed between April 2008 and May 2009. The involved area included L3-S1 in 7 cases, L2-S1 in 3 cases, L3-L5 in 1 cases, L4-S1 in 2 cases. All patients underwent decompression, dynamic stabilization with Bioflex system, according to the severity of degenerative disc with/without interbody fusion. The clinical effects were evaluated by VAS, ODI. ROM and fusion segments were also observed.

RESULTSThe mean follow up period was 19.5 months (from 12 to 26 months). The mean operative time was 183.4 min (from 90 to 240 min) and the mean volume of blood loss was 610.2 ml (from 400 to 1 220 ml). The mean VAS score was 7.8 +/- 1.3 preoperatively, 2.3 +/- 0.9 postoperatively and 2.1 +/- 0.8 at the last follow up. The average ODI was (60.50 +/- 4.40)% preoperatively, (17.80 +/- 2.10)% postoperatively and (16.20 + 2.40)% at the last follow up. The VAS and ODI significant improved in postoperatively (P < 0.05), and there was no statistical difference between postoperative and last follow up (P > 0.05). ROM of whole lumbar and non-fused segment showed obviously decreased and adjacent segment showed insignificant increased. The fusion rate of interbody fusion level was 95.0% (19/20).

CONCLUSIONThe preliminary clinical results show the Bioflex system combined with intebody fusion is a safe and effective technique in treating multi-segment lumbar degenerative disease.

Aged ; Female ; Humans ; Internal Fixators ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; methods ; Spinal Stenosis ; surgery

OBJECTIVETo gain an insight into large genomic deletions in mismatch repair genes MSH2 and MLH1 in Chinese hereditary nonpolyposis colorectal cancer (HNPCC) patients in order to improve genetic detections of HNPCC kindreds.

METHODSFourteen peripheral blood DNA samples were obtained from 14 unrelated HNPCC patients, and fluorescent labeled quantitative multiplex PCR was used to detect large genomic deletions in MSH2 and MLH1 genes.

RESULTSOne of the fourteen probands, a man, was found to have MSH2 exon 1-7 deletions. His cancer-distressed son was also found to have the mutations. Additionally, three normal members of the family had the same mutations.

CONCLUSIONLarge genomic deletions which mainly present to MSH2 account for 20% of general pathological sequence changes of MSH2 and MLH1 genes in Chinese HNPCC patients, and large genomic detections of mismatch repair genes should be included in the regular genetic detections of Chinese HNPCC kindreds.

Adaptor Proteins, Signal Transducing ; genetics ; Asian Continental Ancestry Group ; genetics ; China ; Colorectal Neoplasms, Hereditary Nonpolyposis ; ethnology ; genetics ; pathology ; Female ; Humans ; Male ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; genetics ; Nuclear Proteins ; genetics ; Pedigree ; Polymerase Chain Reaction ; Sequence Deletion

OBJECTIVESTo establish DHPLC method in detecting mutations of mismatch repair genes, hMLH1 and hMSH2, and to identify germline mutations of hMLH1 and hMSH2 in HNPCC kindreds fulfilling Chinese HNPCC criteria.

METHODSFourteen peripheral blood DNA samples from 14 unrelated HNPCC probands fulfilling Chinese HNPCC criteria were obtained respectively. PCR amplified 35 exons of two main mismatch repair genes, hMLH1 and hMSH2. DHPLC followed by DNA sequencing was used to detect and confirm mutations.

RESULTSa total of 41 colorectal cancers and 19 extracolonic tumors were developed in 14 HNPCC kindreds, and gastric cancer was the most common extracolonic tumor type. Twelve single nucleotide changes were identified by DHPLC in 14 probands. Among them, three were missense mutations, one was a nonsense mutation. Other single nucleotide changes included five single nucleotide polymorphisms, two intron single nucleotide changes, one synonymous mutation. hMLH1 EXON19 CODON749 TAC-->TAG (Tyr-->X), hMSH2 EXON12 CODON629 CAA-->CGA (Gln-->Arg) and hMSH2 EXON15 CODON839 CAT-->CGT (His-->Arg) were new discovered mutations.

CONCLUSIONS(1) DHPLC was considered to be highly effective, convenient technique with consistent results for the mutation detection of hMLH1 and hMSH2 genes. (2) Valid mutations of hMLH1 and hMSH2 genes were identified in about one-third HNPCC kindreds fulfilling Chinese HNPCC criteria and missense mutation was the most common mutational types in this cohort of families.

Adaptor Proteins, Signal Transducing ; Asian Continental Ancestry Group ; genetics ; Base Pair Mismatch ; Carrier Proteins ; genetics ; Chromatography, High Pressure Liquid ; Codon, Nonsense ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; DNA Mutational Analysis ; Female ; Genetic Testing ; Germ-Line Mutation ; Humans ; Male ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; genetics ; Mutation, Missense ; Nuclear Proteins ; genetics ; Pedigree

OBJECTIVETo assess the effect of the different treatments targeted at different types of radial head fracture and radial neck fracture.

METHODSA retrospective study was performed in 87 patients from February 2006 to March 2007. Fifty-four patients with radial head fractures included 36 males and 18 females, aged from 18 to 65 years (the average age was 33); Forty of them resulted from crashing, 8 from traffic injury and 6 from falling injury. According to Mason classification system, there were 15 type I, 23 type II and 16 type III. Thirty-three patients with radial neck fractures included 21 males and 12 females, aged from 9 to 17 years (the average age was 13), 29 of them resulted from crashing, 1 from traffic injury and 3 from falling injury. According to O'Brien classification system, there were 8 type I, 14 type II and 11 type III. Type I of radial head fractures and radial neck fractures were immobilization with cast, the patients with type II of radial head fractures were treated with open reduction and micro-screw or T-trapezoid and bridge-shaped plate fixation and type III had operations to fix with bridge-shaped locked plate and repair the broken annular ligament, or replace heads with prosthesis. All patients with type II and type III of radial neck fractures were treated with closed reduction by leverage and percutaneous intra-medullary nailing.

RESULTSThe patients were followed up for 4-12 months (mean 7.2 months). The functional recovery degrees were evaluated with Wheeler's evaluation system. In group of radial head fractures, the results were excellent in 26 patients, good in 20, fair in 6 and poor in 2, the excellent and good rate was 85.2%. In group of radial neck fractures, the results were excellent in 20 patients, good in 9, fair in 4 and poor in no patient, and the excellent and good rate was 87.9%.

CONCLUSIONSDifferent types of fractures should choose different surgical methods according to their characters. The excellent functional recovery depend on anatomical reduction, retaining the head of radius, early repairing and protecting the broken annular ligament of radius, and early functional training.

Adolescent ; Adult ; Aged ; Arthroplasty, Replacement ; Bone Plates ; Bone Screws ; Child ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Radius Fractures ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVESome animal models apply morphine in the drinking water to generate addiction, but related reports are not free of conflicting results. Accordingly, this study aimed to figure out if chronic consumption of morphine in the drinking water can induce morphine addiction in Wistar rats.

METHODSFor 3 weeks, the animals received a daily morphine dose of 35 mg/kg by offering a calculated volume of sugar water (5% sucrose) with morphine (0.1 mg/ml) to each rat; animals receiving just sugar water served as controls. Immediately after the treatment phase, the tail immersion test was used to check for morphine tolerance, and all animals were then kept on tap water for one week (withdrawal phase). Afterwards, all rats were allowed to choose their drinking source by offering two bottles, containing sugar water without and with morphine, simultaneously for two days (preference phase).

RESULTSWhile the chronic consumption of morphine led to a reduction in body weight and to morphine tolerance, the morphine-treated Wistar rats did not show any preference for the opiate-containing sugar water.

CONCLUSIONBody weight loss and tolerance do not reveal a condition of drug craving, and current animal models should be re-evaluated regarding their potential to establish morphine addicted animals.

Animals ; Body Weight ; drug effects ; Choice Behavior ; drug effects ; Disease Models, Animal ; Drug Tolerance ; Male ; Morphine ; administration & dosage ; Morphine Dependence ; Pain Measurement ; drug effects ; Rats ; Rats, Wistar ; Time Factors

OBJECTIVE: To study clinical effects of modified Mclaughlin procedure combined with locking plate for posterior shoulder dislocation with proximal humeral fracture which area of femoral head injury less than 40%. METHODS: From July 2012 to June 2017, 7 patients with posterior shoulder dislocation were treated, including 5 males and 2 females; aged from 37 to 53 years old. Three patients combined with split of humerus head and 4 patients combined with humerus surgical neck fracture. All patients treated with modified Mclaughlin procedure combined with locking plate. Motion of shoulder joint after operation was observed, postoperative UCLA score was used to evaluate clinical effects. RESULTS: Seven patients were followed up from 10 to 33 months. The motion of anteflexion and up-lift ranged from 130° to 170°, the motion of extorsion ranged from 45° to 75°, the motion of abduction ranged fron 105° to 150°, and the internal rotation was between L₃ to buttock. UCLA score ranged from 29 to 34; and 1 patient reached excellent, and 6 patients good. CONCLUSIONS The modified Mclaughlin procedure combined with locking plate showed satisfying result for posterior shoulder dislocation combined with fractures. However, the internal rotation of shoulder could be compromised.
Adult ; Bone Plates ; Female ; Fracture Fixation, Internal ; Humans ; Humeral Fractures ; Male ; Middle Aged ; Shoulder Dislocation ; Shoulder Fractures ; Treatment Outcome