Heparinases,a kind of polysaccharide lyases,can degrade heparin and heparin sulfate to low molecular weight polysaccharides.It has been noted that many bacteria have heparinases although only few of them have been purified and characterized.Heparinases I,II and III from Flavobacterium heparinum have been extensively studied for many years and been commercialized recently.Heparinases have some important applications in the industry and clinic as well as in the determination of heparin structure,which is a very important anticoagulant drug used world-widely.The recent progresses in isolation of heparinase-producing bacteria,genome mapping of heparinase homologs in sequenced bacteria and archaea genomes,purification of heparinases and the study of their biochemistry and regulation were reviewed.The recombinant expression of these enzymes as well as important applications of heparinases and their potential applications in the future will also be highlighted.

Objective To know the expression of 47 kD protein of orientia tsutsugamushi(Ot) Karp strain and test its immunogenicity and antigenicity.Methods Forty-seven kD protein gene that code the whole mat peptide was amplified from the genomic DNA of Ot.Karp strain by PCR and constructed to a recombinant plasmid pGEX-47.The E. coli cells BL21 were transformed with pGEX-47,then the transformants were induced to express the recombinant 47 kD protein by IPTG. The antigenicity and immunogeni-(city) were identified by Dot Blot method and the experiment of immunizing mice respectively.Results 1. The PCR product of 47 kD protein gene of Ot.Karp strain was obtained and its sequence was the same as the published 47 kD protein gene.2. The E.coli expression plasmid pGEX-47 constructed was identified containing the 47 kD protein gene fragment by restrication analysis and PCR identification,and an expression band about 47 kD was detected by SDS-PAGE of the purified product from GST-fusson protein expressed by pGEX-47. 3.Dot Blot analysis and the experiment of immunizing mice testified its antigenicity and immunogenicity.Conclusions The 47 kD protein gene of Ot Karp strain is amplified that code the whole mat peptide, which can be expressed in prokaryotic cells and the expressed protein has antigenicity and immunogenicity.

Objective To study the way of quick identification of haemophilus influenzae with reverse dot blot.Methods Oligonucleotide probe which is specially targeted to 16SrDNA of haemophilus influenzae was designed, then fixed the probe to nylon membranes, and hybridized with the production of gain with the universal primers.Results The universal primers could hybridize the target sequence from common pathogenic bacteria by PCR, and oligonucleotide probe could hybridize with haemophilus influenza specially and could not hybridize with other bacterias. It proved that the probe was of highly speciality.Conclusion Reverse dot blot is a good method of quickly identification of haemophilus influenzae.

This paper established a near-infrared spectroscopy quantitative model for simultaneous quantitative analysis of coptisine hydrochloride, dehydrocorydaline, tetrahydropalmatine, corydaline and glaucine in Corydalis Rhizoma. Firstly, the chemical values of the five components in Corydalis Rhizoma were determined by the reversed-phase high performance liquid chromatography (RP-HPLC) with UV detection. Then, the quantitative calibration model was established and optimized by fourier transformation near-infrared spectroscopy (NIRS) combined with partial least square (PLS) regression. The calibration model was evaluated by correlation coefficient (r), the root-mean-square error of calibration (RMSEC) and the root mean square of cross-validation (RMSECV) of the calibration model, as well as the correlation coefficient (r) and the root mean square of prediction (RMSEP) of prediction model. For the quantitative calibration model, the r, RMSEC and RMSECV of coptisine hydrochloride, dehydrocorydaline, tetrahydropalmatine, corydaline and glaucine were 0.941 0, 0.972 7, 0.964 3, 0.978 1, 0.979 9; 0.006 7, 0.003 5, 0.005 9, 0.002 8, 0.005 9; and 0.015, 0.011, 0.020, 0.010 and 0.022, respectively. For the prediction model, the r and RMSEP of the five components were 0.916 6, 0.942 9, 0.943 6, 0.916 7, 0.914 5; and 0.009, 0.006 6, 0.007 5, 0.006 9 and 0.011, respectively. The established near-infrared spectroscopy quantitative model is relatively stable, accurate and reliable for the simultaneous quantitative analysis of the five alkaloids, and is expected to be used for the rapid determination of the five components in crude drug of Corydalis Rhizoma.
Alkaloids ; chemistry ; Corydalis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Rhizome ; chemistry ; Spectroscopy, Near-Infrared ; methods

OBJECTIVETo explore methods of treating middle and distal tibia nonunion with the treatment of advanced bone graft combined with locking compression plate.

METHODSFrom January 2011 to December 2012, 12 patients with middle and distal tibia nonunion were treated with advanced bone graft combined with locking compression plate. Among patients, there were 8 males and 4 females aged from 20 to 69 with an average of 47 years old. The time from first injuries to bone nonunion was from 9 months to 5 years, avergaed 19 months. Four cases were treated with external fixation, 6 cases were treated with plate fixation, 2 cases of 12 patients occurred broken of plate and nail. Eleven patients were non-infective bone nonunion and 1 patient was infective bone nonunion. Preoperative X-ray and CT showed all patients had sequestration and formation of ossified bone with different degrees. Operative time, blood loss, wound healing were observed, fracture healing time was evaluated by postoperative X-ray. Johner-Wruhs scoring standards was used to evaluate ankle joint function after operation at 10 months.

RESULTSOperative time ranged from 90 to 185 min with an average of (125.00±20.15) min; blood loss ranged from 225 to 750 ml with an average of (415.00±120.00) ml. All patients were followed up from 10 months to 2.5 years with an average of 1.5 years. Postoperative X-ray showed bone union was formed around fracture after operation at 4 months in all patients, 3 cases obtained bone healing within 6 months after operation, 9 cases obtained from 8 to 12 months. No infection, injury of nerve and vessles, and broken of plate and nail were ocurred. According to Johner-Wruhs scoring at 10 months after operation, 10 cases obtained excellent results, 1 good and 1 moderate.

CONCLUSIONAdvanced bone graft combined with locking compression plate, which can build fracture multi-point supporting based on full compression of bone nonunion to get effective fixation, is an effective method in treating middle and distal tibia nonunion.

Adult ; Aged ; Bone Plates ; Bone Transplantation ; Female ; Fracture Healing ; Fractures, Ununited ; surgery ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery

Follicle-stimulating hormone (FSH) is synthesized and secreted by the anterior pituitary, which binds to its receptors expressed on the membrane of Sertoli cells in the testis to bring about spermatogenesis. With the development of DNA sequencing technology, FSH SNPs rs10835638 and FSHR SNPs rs6165, rs6166, and rs1394205 were detected, which might directly affect the expression of FSH and activity of FSHR, resulting in male spermatogenic dysfunction. This review focuses on the relationship of FSH and FSHR gene polymorphisms with male infertility.
Follicle Stimulating Hormone ; genetics ; Humans ; Infertility, Male ; genetics ; Male ; Polymorphism, Single Nucleotide ; Receptors, FSH ; genetics ; Sertoli Cells ; Spermatogenesis ; Testis

Objective To establish the HPLC fingerprint of Sanhuang Xiexin Decoction and provide a method to study the potential basis and the changing of the chemical component for it in different compati- bility.Methods An HPLC method was established with Shimadzu ODS column(150 mm?4.6 mm, 5?m),the mobile phase was methanol-water-0.1% phosphoric acid(0.01 mol/L potassium phosphate monobasic,pH 2.8)as gradient elution,the flow rate was 1.0 mL/min,and the detection wavelength was 260 nm.Through comparing and analyzing the relative retention time of this decoction and of its composi- tion which are positive and negative control fingerprints,the main chromatographic peak origins were con- firmed;The correlated chromatographic peaks were identified by contrasting chromatographic peak reten- tion time and adding reference substances to the sample.Results All tested samples contained the 32 common peaks,the relativity of them were analyzed and 11 peaks were indicated.The similarity of ten batches of samples exceeded 0.92.Conclusion This method shows sensitive and good repeatability,all of the contents are separated well.It is used to determine Sanhuang Xiexin Decoction and its relative prepara- tions.

This study investigated the mental health status of medical students in China, and analyzed the influencing factors in order to provide evidence for mental health education for medical students. A stratified cluster sampling method was used to recruit medical students from Huazhong University of Science and Technology, China. The questionnaire survey on general information and Symptom Checklist 90 (SCL-90) were used for investigation and analysis. The results showed among the 1137 valid questionnaires, 278 (24.45%) participants had SCL-90 score ≥ 160. The top three mental problems of medical students were obsessive-compulsive disorder, interpersonal sensitivity and depression in terms of the factor score ≥ 2.5 and the number of participants who reflected on the diseases. The third-year medical students had the worst mental health status, and fifth-year medical students had the best mental health status. Students from rural area had more psychological problems than those from urban area; furthermore, students with high professional satisfaction, those who were the single child of the family, non-poor students, and those whose parents had high education level had better mental health status. It was concluded that the mental health of medical students is not optimistic in China. Medical students have some mental health problems of different degrees. Factors that influence the mental health of medical students include academic pressure, professional satisfaction level and family environment.

The main treatment strategies for chronic pancreatitis in young patients include therapeutic endoscopic retrograde cholangio-pancreatography (ERCP) intervention and surgical intervention. Therapeutic ERCP intervention is performed much more extensively for its minimally invasive nature, but a part of patients are referred to surgery at last. Historical and follow-up data of 21 young patients with chronic pancreatitis undergoing duodenum-preserving total pancreatic head resection were analyzed to evaluate the outcomes of therapeutic ERCP intervention and surgical intervention in this study. The surgical complications of repeated therapeutic ERCP intervention and surgical intervention were 38% and 19% respectively. During the first therapeutic ERCP intervention to surgical intervention, 2 patients developed diabetes, 5 patients developed steatorrhea, and 5 patients developed pancreatic type B pain. During the follow-up of surgical intervention, 1 new case of diabetes occurred, 1 case of steatorrhea recovered, and 4 cases of pancreatic type B pain were completely relieved. In a part of young patients with chronic pancreatitis, surgical intervention was more effective than therapeutic ERCP intervention on delaying the progression of the disease and relieving the symptoms.

· AIM: To explore the dynamic expression and correlation among telomerase catalytic subunit (TERT), proliferating cell nuclear antigen ( PCNA) and antiapoptosis protein Bd-2 which relate to cell proliferation in epiretinal membrane of rat traumatic proliferative vitreoretinopathy(PVR).· METHODS: S-P technique was applied for immunohistochemical staining of epiretinal membrane of traumatic PVR with TERT, PCNA and Bcl-2 antibody. HE staining was also carried out. The staining results were analyzed with image analysis system.· RESULTS: The positive rate and average A of PCNA protein were upregulated at first and then down-regulated, with the peak value in 14d Group, which was significantly different from those in 7d Group and 28d Group.The positive rate and average A of TERT and Bcl-2 were also upregulated at first and then down-regulated, with the peak value in 14d Group and 21d Group, which were significantly different from those in 7d Group. There was significant correlation among PCNA, Bcl-2 and TERT protein expression (P≤0.01).· CONCLUSTON: TERT and Bcl-2 take part in the regulation of proliferative cells in epiretinal membrane of traumatic proliferative PVR, with high correlation with the dynamic changes of cell proliferation.