Objective To express the major allergen of Blattella germanica (Bla g 2) in Pichia pastoris and obtain the soluble protein. Methods The known Bla g 2 gene was used to design the primers which had the restriction enzyme sites. PCR method was applied to obtain the Bla g 2 gene. The gene fragment was then cut and ligated with the Pichia expression vector pGAPZaA, resulting in a recombinant plasmid pGAPZaA-Bla g 2. The linearized pGAPZaA-Bla g 2 was transformed into Pichia pastoris GS115 through electroporation, then screened to positive transformants, and the protein was expressed in YPD medium. Purification of the recombinant protein was achieved by metal (Ni2+) chelating affinity chromatography and Western-blotting assay indicated its IgE binding capacity. Results With the expressed reeombinanl protein, SDS-PAGE showed the presence of the product in the supernatant of the culture with Mr 45 000. After 3 days culture, the recombinant protein occupied 50% of the total proteins in the supernatant. The recombinant protein was purified and Western-blot demonstrated an adequate IgE binding capacity of the product. Conclusion A recombinant protein of Bla g 2 has been obtained, which is soluble in the supernatant and therefore can avoid a process of denaturalization and renaturation of the recombinant.

This paper dicusses the significance of drug clinical trials,and summarises the experiences in policy making,personnel trainning,supervising and management in drug clinical trials at the First People's Hospital of Changzhou,which may provide reference for drug clinical trials in China.

Objective To compare the difference of clinical effect on the thoracic trauma patients between conventional thoracotomy(CTH) and video-assisted thoracoscopic surgery (VATS), and the possible mechanism.Methods Two hundred thoracic trauma patients were enrolled from 2007 January to 2010 March,and divided into VATS group( n =98) and CTH group( n = 102) according to the operative methods. The levels of serum high sensitive C-reactive protein(hs-CRP) ,interleukin-6 (IL--6) and tumor necrosis factor-α (TNF-α) were determined before and after operation. Intraoperative blood lost, operative time, postoperative analgesic time and hospitalization time were recorded and analyzed. Results There were no differences on the levels of inflammatory factors between the two groups before operation(P＞0.05) ,but the levels of inflammatory factors of VATS group were significantly decreased than that of CTH group after operation ( all P < 0. 05). Intraoperative blood lost, operative time, postoperative analgesic time and hospitalization time of VATS group were also lower than that of CTH group ( all P < 0. 05 ). Conclusion Compared with conventional thoracotomy, video-assisted thoracoscopic surgery had the characteristics of less intraoperative blood lost, shorter operative and postoperative analgesic time, and rapid recovery, could be relative to less inflammatory reaction.

The nature and preparation of sodium alginate are introduced and its medical applications are reviewed in fields such as medical carrier,gel microspheres,sustained-release materials,surgical wound repairing materials,and so on.Besides,its development prospects are described.It is an inevitable trend that sodium alginate is used in the development of controlled-release reagent.There are also important applications in the partial treatment of ear disease and artificial cartilage for sodium alginate,but the strength and toughness of alginate hydrogel limits its wider application.

Objective To discuss the preoperative diagnosis and the effect of microsurgical transsphenoidal surgery of pituitary Rathke's cleft cysts.Methods Retrospective analysis was performed from January,2011 to May,2015 on 62 cases of Rathke's cleft cyst which confirmed by surgery and pathology at the First Affiliated Hospital of Sun Yat-sen University.Sixty-two cases were performed by the surgery of transsphenoidal approach.Results There were 50 cases with a correct preoperative diagnosis of and consider Rathke's cleft cyst,12 cases of misdiagnosis.Postoperative follow-up was performed within 6-12 months,and the patient's clinical symptons were improved in different degrees,1 case of recurrence,no deaths and serious complications.Conclusion The diagnosis of Rathke's cleft cyst need for comprehensive considerations from many aspects,and the improvement of diagnosis rate base on the clinical features,endocrine examination,and imaging data.The microsurgical transsphenoidal surgery is safe and effective treatment for Rathke's cleft cyst.

Metastasis-associated in colon cancer-1 (MACC1) is a recently discovered gene associated with colon cancer metastasis,there is significant relationship indicated from some studies between MACC1 and different malignant tumors.It may play an important role in the regulation of tumors metastasis.This article reviewed the expression and regulating function of MACC1 in different cancers including colorectal cancer,hepatic cancer,gastric cancer,lung cancer,ovarian cancer,breast cancer,and so on.It may offer clues to find a new target for target treatment of cancer metastasis.

The main reasons to doubt the validity of serum pharmacology include:①The components of medical herbs absorbed from gastrointetinal tract enter circulating plasma,not serum in vivo.②Blood coagulation involves the activation of a series of proenzymes,and the proteases induce leukocytes to release lysosomal enzymes.These enzymes may degrade some herbal components.③Thrombin can stimulate platelets and leukocytes to secrete many biologically active substances.④Fibrinolysis,complement and kinin systems are activated during blood coagulation.⑤The processes of serum preparation and inactivation always cause lose of lots of herbal components. Recently finger printing analysis using HPLC-MS and antileukemic proliferation experiments showed that the results obtained from serum pharmacological method might be incorrect. We propose that the ex vivo experiments on multiple levels are necessary to elucidate the pharmacological actions and their mechanisms of medical herbs and believe that the pharmacological method using plasma absorbed herbal components should be better than that using serum.

Aim To establish an ideal hyperlipidemia animal model by a new way.Methods 30 mice were randomly divided into control group, positive control group and model group.The mice of control group were only fed with a standard diet.Those of positive control group were fed with high fat diet.Those of model group were fed with a standard diet and enough milk.After 30 days the TG, TC, HDL-C and LDL-C of serum and hepatic and LI were detected and pathological changes in the liver of mice were observed microscopically.Results Compared with the control group, the mice of model group developed hyperlipidemia with LI and the serum and the hepatic TG, TC, LDL-C elevated significantly, while HDL-C were significantly lower.The histopathological research showed hepatocellular macrovesicular steatosis and hepatitis in the model group.Conclusion An ideal model of hyperlipidemia is successfully established with standard diet and milk fed to mice for 30 days.

BACKGROUND: It has been confirmed that some casting alloy can inhibit the activity of human gingival fibroblasts in vitro, which can be used for inducing apoptosis of human gingival fibroblasts (HGFs). However, the mechanism of this effect is poorly understood.OBJECTIVE: To investigate the effect of Ni-Cr, Co-Cr, Au and pure Ti ceramic alloys on the expression of P70S6k by HGFs in vitro.DESIGN, TIME AND SETTING: Grouping controlled experiments with the expression of P70s6k by HGFs in each group as observation objects. Experiments were performed at the Institute of Immunohistochemistry and Western blotting, Technology Building of Liaoning Medical University from July to October 2009.MATERIALS: Ni-Cr, Co-Cr, Au and pure Ti ceramic alloys were processed by Shenzhen Mecodent Dental Laboratory Co., Ltd. The gingival from gingival neck of the first premolar (which goes to be extracted due to the need of orthodontic treatment) of teenager who was15 years old without any clinical disease was obtained, followed by the primary culture of HGFs. The fifth passage of cells in logarithmic phase were used to measure indexes.METHODS: Ni-Cr, Co-Cr, Au and pure Ti ceramic alloys were incubated in DMEM to prepare alloy leaching liquor, which were then added in HGF medium at 10% concentration. DMEM containing 10% fetal bovine serum was used in negative controls. MAIN OUTCOME MEASURES: The expression of P70S6k in each group was measured by Western Blotting following 72 hours of intervention with alloy leaching liquor.RESULTS: Western-blot results showed that there was no significant difference in the average gray value of P70S6k expression in Au, pure Ti ceramic alloys and control groups (P > 0.05). There was also no significant difference between Au ceramic alloys and pure Ti ceramic alloys groups or between Ni-Cr ceramic alloys and Co-Cr ceramic alloys groups. (P > 0.05), but Ni-Cr ceramic alloys group or Co-Cr ceramic alloys group showed significant difference compared with the Au, pure Ti ceramic alloys and control groups (P < 0.01).CONCLUSION: Au ceramic alloys and pure Ti ceramic alloys show that they have no obvious effect on the proliferation activity of HGFs. Ni-Cr and Co-Cr ceramic alloys showed an inhibitory effect on the proliferation activity of HGFs.

Some grain sprouts herbs recorded in Chinese Pharmacopoeia 2010, including Oryzae Fructus Germinatus, Setariae Fructus Germinatus and Hordei Fructus Germinatus, together with their adulterants, were identified by ITS2 sequences. The genomic DNA were extracted, their ITS2 sequences were amplified, and purified PCR products were
sequenced. Sequences were assembled using the CodonCode Aligner. The genetic distances, variable sites and the neighbor-joining (NJ) phylogenetic tree were computed by MEGA 6.0 in accordance with the Kimura 2-Parameter (K2P) model. The results showed that their intraspecific genetic distances were all much lower than the interspecific ones of themselves and their adulterants. The NJ tree based on ITS2 sequence indicated that Oryza sativa, Setariaitalica, and Hordeum vulgar formed one monophyletic clade respectively, in addition any one of the grain sprouts herbs and its adulterants could be distinguished clearly. Therefore, ITS2 sequence is suitable to be as a barcode to identify the grain sprouts herbs and their adulterants.