Chitinases genes from Metarhizium anisopliae which is an important entomopathogenic fungus were considered one of the key factors to invade their hosts. One Metarhizium anisopliae HN1 strain was isolated and screened. A chitinase gene was amplified by RT-PCR from Metarhizium anisopliae HN1, The whole length of this gene was 1275bp,and the nucleotide sequence of the gene was 96% similarity to that of the M. anisopliae E6 accessed in GenBank ( AF02749). The gene has been registered in GenBank and its accession number is DQ011865. The gene was subcloned into prokaryon expression vector pET-22b( + ), transformed this recombinant expression plasmid into E. coli strain BL 21 and effective expressed. The SDS-PAGE analysis indicated that the recombinant protein was 42kDa which is same to the reported article. The expression level of recombinant protein was about 63. 3% of whole expressed proteins , And when recombinant E. coli were crushed by freeze and supersonic wave , the activity assay indicates that the chitinase expressed in bacteria possesses biological activity.

Procedure for prolapse and hemorrhoids( PPH) is one of the important techniques developed for the treatment of hemorrhoids with severe degree in the last decade. Its principle is based on the "anal cushion" theory.Compared with traditional hemorrhoidectomy , PPH has advantages of shorter operation time , minor degree of postoperative pain , shorter hospital stay and quicker recovery.However, the occurrence of relapse and re-prolapse of hemorrhoids is high. Besides, the short-term efficacy of PPH for the constipation outlet obstruction caused by anterior rectocele is also favorable.
Anal Canal ; Hemorrhoids ; diagnosis ; Humans ; Operative Time ; Pain, Postoperative ; Prolapse ; Surgical Stapling

This study is to establish the fingerprint for Phyllanthus emblica and their tannin parts from different habitats by HPLC for its quality control. The determination was carried out on a Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column, with methanol-0.2% glacial acetic acid as mobile phase with gradient elution at a flow rate of 1 mL x min(-1). The temperature was maintained at 30 degrees C and the detected wavelength is 260 nm, Thirteen chromatographic peaks were extracted as the common peaks of the fingerprint of P. emblica, and eleven as the common peaks of P. emblica tannin parts, and five peaks were identified by comparing with referent samples. The fingerprints of 8 samples were compared and classified by similarity evaluation, cluster analysis and principal component analysis (PCA). The similarity degrees of eight P. emblica were between 0.763 and 0.993, while tannin parts were between 0.903 and 0.991. All the samples of P. emblica and their tannin parts were classified into 3 categories. The method was so highly reproducible, simple and reliable that it could provide basis for quality control and evaluation of P. emblica from different habitats.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Medicine, Tibetan Traditional ; Phyllanthus emblica ; chemistry ; classification ; Quality Control ; Tannins ; analysis ; Tibet

OBJECTIVETo investigate the effect of pyrroloquinoline quinone (PQQ) on nerve regeneration of transected sciatic nerve in animal models.

METHODSForty SD rats weighing 220-240 g were randomized into a PQQ group (n = 20) and a control group (n = 20). Each animal underwent sciatic nerve transection operation. After the operation, PQQ 0.5 ml (250 microg/Kg) was injected at the operation site in the PQQ group, while the same volume of normal saline was delivered in the control group. Nerve functional evaluation, electrophysiological index recording were carried out according to the experimental design. Newly generated nerve specimens were harvested 12 weeks postoperatively for morphological studies.

RESULTSIn the PQQ group there was a good nerve regeneration and the sciatic nerve function, sciatic nerve function index, electrophysiological index and morphological appearance were superior to the control group (P < 0.05).

CONCLUSIONSPQQ has a remarkable effect in enhancing nerve regeneration of transected peripheral nerve.

Animals ; Male ; Nerve Regeneration ; drug effects ; PQQ Cofactor ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; pathology ; physiology

Objective To investigate plasma glutathione S-transferase(GSTs) activity and GSTP1 gene Ile105Val polymorphism in Bijie City, Guizhou Province, a coal-burning fluorosis endemic area. Methods One hundred and sixty villagers from Yachi Twon using non-improved cooking stoves were selected as the non-intervened group in Bijie City, Guizhou Province where coal-burning fluorosis was prevailing; 153 villagers as the intervented group were chosen from Changchun Twon, where cooking stoves were improved; 151 villagers were served as the control group from Baiyunshan Twon, Changshun County without endemic fluorosis. The activity of GSTs was tested by colorimetric analysis with spectrophotometer. The genotype of the GSTP1 gene Ile105Val polymorphism, presenting as either homozygous wild-type (AA), or heterozygous mutation type (AG), or homozygous mutation type (GG), was detected through the PCR-RFLP procedure. Results The activity of GSTs in plasma of non-intervened group [(12.44±4.97) kU/L]was significantly lower than that of intervened group (P < 0.05), and that of intervened group[(20.78±6.20)kU/L]was significantly lower than that of control group[(24.30±6.27)kU/L, P< 0.05]. The difference of the enzyme activity of three groups were statistically significant (F = 51.71, P < 0.05), but this enzyme activity did not vary significantly in each sex of each grnup(P > 0.05). Compared intervened group [AA:67.3%(103/153), AG:29.4%(45/153),GG:3.3%(5/153)]and non-intervened group[AA:66.9%(107/160), AG:30%(48/160), GG:3.1%(5/160)]with control group[AA:74.8%(113/151), AG:25.2%(38/151), GG:0 (0/151)], the Ile105Val polymorphism site of GSTP1 gene had significant difference(χ2= 6.04,6.07, both P< 0.05), but not significant between intervened and non-intervened groups(χ2 = 0.02, P>0.05). Conclusions Fluorosis can decrease the activity of GSTs and introduce the GSTP1 gene Ile105Val polymorphism, intervention with the fluorine intake will improve the effect of fluoride on the body.

[Objective]To investigate the cytotoxic effect ofaquaporin-4 (AQP4) antibody onin vitro cultured rat cortical neurons.[Methods]Cortical cells were obtained from Wistar embryonic rats with 16-19-d pregnancy;3 daRer seeding,the cultured cortical cells were randomly divided into a control group and an AQP4 antibody-positive serum treatment group.Neurons of the control group were added the normal human serum and those in the treatment group were added serum from AQP4 antibody positive patients.The number and morphology changes of astrocytes,neurons and microglias were observed by immunohistochemistry at 2,4 and 6 h after being incubated with AQP4 antibody positive serum or controls.[Results] No number or morphology changes of cortical cells were observed in control group at different time points.AQP4 antibody-positive serums induced astrocyte swelling,microglia hyperplasia and neuron axonotmesis,but did not influence the numbers of cortical cells at 2 h after induction.The numbers of astrocytes and neurons in the treatment group (24.73％±5.27％ and 35.49％±8.43％,respectively) significantly decreased as compared with those in the control group (30.34％％:t,4.53％ and 48.60％+10.99％) at 4 h after induction (P＜0.05),while those of microglias in the treatment group (27.35％±13.17％) obviously increased as compared with those in the control group (16.44％±2.70％,P＜ 0.05);these changes became more significant at 6 h after induction.[Conclusion] AQP4 antibodies induce loss of astrocytes and neurons,and microglia activation in primary cortical cells,which maybe imply the primary pathogenic role of AQP4 antibody in neuromyelitis optica.

OBJECTIVETo investigate the prevention and treatment of complications during and after endoscopic mucosal band ligation (EMBL) for precancerous lesions and early cancer in the esophagus.

METHODSClinical data of 47 patients with esophageal precancerous lesions and early cancer undergoing EMBL in our center from June 2011 to August 2013 were reviewed retrospectively. Complications and associated treatment during operation, after operation and during follow-up were analyzed.

RESULTSComplications during operation included 7 cases of bleeding (14.9%) and 1 case of perforation (2.1%), who received hot biopsy forceps and argon plasma coagulation to stop bleeding successfully, and titanium clamp to suture wound surface. No cutaneous emphysema and pneumothorax occurred. Complications after operation included 1 case of delayed bleeding (2.1%) who received blood stopping under gastroscope, 2 cases of mediastinal and subcutaneous emphysema (4.3%), 6 cases of pleural effusion (12.8%), and 5 cases of minor inflammation or segmental atelectasis of pulmonary (10.6%), who all received successful conservative treatment. Seven cases of esophageal stricture occurred during follow-up, who were improved by balloon dilatation and metal-film stent placement. No deaths associated with EMBL occurred. All the complications were cured through conservative treatment. No additional surgery associated with the complications was needed. Post-operative pathology revealed 1 case was chronic inflammatory hyperplasia, 11 were low-grade intraepithelial tumor, 15 were high-grade intraepithelial tumor, 8 were carcinoma in situ, 12 were squamous cancer (8 with invasion into mucous muscular layer, 4 into submucous layer). Only 1 case of submucous cancer needed transthorax esophageal cancer radical operation because of dangerous margin. No relapse case was found during followed-up.

CONCLUSIONEMBL can treat the esophageal precancerous lesions and early esophageal cancer effectively and its complications can be managed with conservative therapy usually.

Adult ; Aged ; Aged, 80 and over ; Endoscopy ; adverse effects ; Esophageal Neoplasms ; surgery ; Female ; Humans ; Ligation ; Male ; Middle Aged ; Mucous Membrane ; surgery ; Postoperative Complications ; prevention & control ; Precancerous Conditions ; surgery ; Retrospective Studies

OBJECTIVETo investigate the effects of rhVEGF on autologous free granular fat grafts in rats.

METHODSForty-eight Sprague-Dawley rats were randomly divided into three groups, sixteen of each. After the autologous free granular fat transplantation, all groups were treated with the plasmid DNA containing cDNA encoding rhVEGF, the blank plasmid DNA and normal saline respectively as the experimental group, the negative group and the saline group. After 3, 7, 15, 30 days, the rats were sacrificed and the grafts were weighted accurately. Histological pathology was evaluated. Micro-vessel count and the expression of vascular endothelial growth factor (VEGF) were examined by immunohistochemical staining.

RESULTSThe weights of the two latter groups were significantly reduced on the 7, 15, 30 day compared with the experimental group. The expression of VEGF and the micro-vessel count in the experimental group were significantly higher than the other two groups during the latter periods.

CONCLUSIONThe cDNA encoding VEGF can induce the expression of VEGF in fat graft, angiogenesis and reduce the free fat graft absorption.

Adipose Tissue ; transplantation ; Animals ; Female ; Gene Transfer Techniques ; Graft Survival ; Male ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; genetics ; Transplantation, Autologous ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo investigate the effects of peroral endoscopic myotomy(POEM) on esophageal dynamics in patients with esophageal achalasia.

METHODSFrom September 2011 to November 2011, 20 cases with esophageal achalasia received POEM at the Endoscopic Center in the Zhongshan Hospital of Fudan University. Pre-operation esophageal dynamics of all the patients were evaluated by high resolution manometry(HRM) system and 3 days after operation the test was repeated. Lower esophagus sphincter resting pressure(LESP), 4-second integrated relaxation pressure(4sIRP), lower esophagus sphincter relax rate(LESRR), lower esophagus sphincter length(LESL), and esophageal manometry were analyzed.

RESULTSAfter POEM, LESP decreased from(29.1±17.0) mm Hg to(14.6±4.9) mm Hg, and decrease rate was 49.8%(P<0.01). However, the decreases in LESRR and LESL were not statistically significant(P>0.05). Esophageal peristaltic contraction was absent in all the 20 patients preoperatively. After POEM, changes in the esophageal contraction were seen in 7 patients, and peristalsis was noticed but was below normal level. There were no significant changes in peristalsis in the remaining 13 patients.

CONCLUSIONPOEM can significantly reduce LESP and 4sIRP in patients with achalasia, but can not affect the contraction of the esophagus.

Adolescent ; Adult ; Aged ; Esophageal Achalasia ; physiopathology ; surgery ; Esophagoscopy ; methods ; Esophagus ; physiopathology ; surgery ; Female ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo explore the dynamics of hepatitis B virus covalently closed circular DNA (cccDNA) and optimal duration of treatment after serum virology response.

METHODSHBV cccDNA in liver biopsies and the serum HBV DNA were quantified by real time PCR, the serum makers were detected by enzyme-linked immunosorbent assay.

RESULTS(1) The cccDNA in biopsy samples continued to decrease after serum virology responded. (2) The longer the treatment after serum virology response, the lower the cccDNA level in liver tissue. (3) Anti-HBe positive patients had lower cccDNA in liver tissue than anti-HBe negative patients. (4) cccDNA in liver tissue was undetectable in 12 out of the 18 case anti-HBe(+) patients. Serum virology response lasted 35 months and anti-HBe(+) lasted 30 months.

CONCLUSIONAfter serum virology responded, the longer the treatment, the lower the liver cccDNA. The cccDNA is undetectable in about 2/3 of the patients if the serum virological clearance lasts more than 35 months and anti-HBe lasts more than 30 months.

Adult ; Aged ; Antiviral Agents ; therapeutic use ; Biopsy ; DNA, Circular ; analysis ; DNA, Viral ; analysis ; blood ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Liver ; pathology ; virology ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Time Factors ; Viral Load ; Young Adult