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.

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.

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.

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.

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.

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.

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.

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.

Objective To evaluate the clinical efficacy of intracavitary holmium laser for ureter calculi combined with distal ureteral stricture.Methods Sixty-nine patients with ureteral calculi combined with distal ureteral stricture were selected.Nineteen patients with ureteral calculi above third lumbar vertebra inferior margin combined with distal ureteral stricture were given microchannel percutaneous nephroscope holmium laser lithotripsy + ureteral stricture dilatation and holmium laser incision,and 50 patients with ureteral calculi below third lumbar vertebra inferior margin combined with distal ureteral stricture were given ureteroscope holmium laser lithotripsy + ureteral stricture dilatation and holmium laser incision.The type-B ultrasonic,kidney-ureter-bladder plain film (KUB) + intravenous urography (IVU) were performed before operative and 3 months after removal of double J tube,and the degree of hydronephrosis and calculi removal condition were observed.Results The operations of all the patients were successful,and operation time was 42-85 (58 ± 13) min.Intraoperative hemorrhoea,ureteral rupture,ureteral avulsion,false passage formation did not occur.All patients had varying degrees of gross hematuria after operation,and 68 cases disappeared after 2-3 d.Thirteen cases had postoperative fever,and these patients were alleviated after the anti-infection and symptomatic treatment.The success rate of gravel was 95.7％ (66/69),and the success rate of ureteral stricture was 97.1％ (67/69).Three months after removal of double J tube,calculi had been discharged,with no residual calculi.The patients' hydronephrosis was obviously alleviated.The extension of renal collecting system was (22.0 ± 8.2) mm before operation,and was (12.0 ± 6.1) mm 3 months after extraction double J tube,and there was statistical difference (t =8.52,P ＜ 0.01).Conclusions The nicrochannel percutaneous nephroscope holmium laser lithotripsy or ureteroscope holmium laser lithotripsy + ureteral stricture dilatation and holmium laser incision for ureteral calculi combined with distal ureteral stricture patients has no serious intraoperative and postoperative complications,shorter operation time,low cost,operation safety,and satisfactory effect.It is a good method for ureteral calculi combined with distal ureteral stricture.

Objective To investigate the safety and efficacy of flexible ureteroscopy and holmium laser lithotripsy for the treatment of renal stones in patients with a solitary kidney.Methods Between May 2014 and June 2016,Clinical records of 17 cases of stones in a solitary kidney who underwent flexible ureteroscopy and holmium laser lithotripsy were reviewed.10 of these cases were male and the others female;mean age of these cases was 45.8 (32-67) years.9 were on the left side while 8 were on the right.6 cases were with functional solitary kidney (with the GFR of the contralateral kidney less than 10ml/min),while 11 others were with anatomical solitary kidney (6 due to tumor,2 due to tuberculosis,3 being congenital).All cases underwent BUS,KUB as well as CTU to confirm diagnosis before surgery,with 8 diagnosed with renal pelvis calculi,5 were with upper or middle calyx calculi,and 4 were with subrenal calyx calculi.The mean ± SD size of the stones was 1.2 ± 0.3 cm.Clinical m anifestations were asymptomatic in 10cases,backache in 5 cases and fever in 2 cases.All patients underwent exams of CBC,urinalysis,liver and kidney function,coagulation function,ECG,chest X-ray as well as cleaning midstream urine cultivation after admission.Results Mean ± SD operation time among all cases was (74.2 ± 23.6) min,the amount of bleeding during surgery was 5-15 (mean:8) ml,duration of hospitalization was 3-12 (mean:4.5)days,ureteral stents are kept for an average time of 4 weeks after surgery.3 patients developed fever after operation,1 developed urosepsis;all these patients recovered after active anti-infection treatment.No post-operative complications such as fever,renal colic,hematuresis,ureteral perforation,active bleeding as well as sepsis were found among the other patients.The preoperative and postoperative serum creatinine had statistical difference (P ＜0.05).14 patients were proved to be stone-free by KUB or CTU.The stone-free rate after a single operation was 82.3％ (14/17).Residual stone were found in the 3 patients,who subsequently underwent a second flexible ureteroscopy and holmium laser lithotripsy.The stone-free rate after a second operation was 94.1％.Conclusions Flexible ureteroscopy and holmium laser lithotripsy is a safe and effective method for the treatment of nephrolithiasis in patients with a solitary kidney with a lower procedure-related injury rate,smaller amount of bleeding,a lower complication rate and a shorter time required to recover after surgery.