The major challenge in the development of recombinant biologics lies in generating and isolating rare high-producing stable single clone in a short period of time. The selection marker is an essential component of the plasmid vector, it plays an important part in the generation and screening of producing cell lines. Engineering the selection marker to enhance the stringency of selection for high producing cells is one of the most effective approaches to improve the cell line development process. Here, using Chinese hamaster overy (CHO) cells as an example, we introduce the application of selection marker for generation of recombinant biologics producing mammalian cell lines, methods of engineering the selection markers to enhance the selection stringency, and propose considerations on cell substrate stability and selection marker safety, in order to provide references for high-efficiency development of recombinant biologics.

OBJECTIVE To investigate the clinical effect of Ashi Dispel-itch solution on treatment of diabetic-candidal vaginitis.METHODS Totally 322 patients were randomly divided into 2 groups,one group was treated with Ashi Dispel-itch solution by fumigating and soaking ill section while the control group with Jieyin solution.The curative effect was divided into 4 grades according to symptoms,body appearance,level of pH,etc.RESULTS The total effective rate due to Ashi Dispel-itch solution was 93.5% and was significantly higher than that of Jieyin solution(P

Adult ; Alkanesulfonic Acids ; adverse effects ; Humans ; Male ; Pemphigoid, Bullous ; chemically induced

Angiolymphoid Hyperplasia with Eosinophilia ; pathology ; Forehead ; pathology ; Humans ; Male ; Middle Aged ; Parotid Gland ; pathology

OBJECTIVE:To study the anti-tumor effects of Vinblastine (VLB) hydrophilic group modified cationic liposomes in tumor-bearing mice. METHODS:Tumor-bearing model were induced by inoculating yellow ascites of S180 ascites tumor mice. Tumor-bearing mice were randomly divided into model group,VLB sulfate injection group,VLB liposomes group,VLB hydrophil-ic group modified liposomes group,VLB cationic liposomes group and VLB hydrophilic group modified cationic liposomes group, i.e. group A,B,C,D,E and F,with 18 mice in each group. Group A was given normal saline intravenously via mice tail,other groups were given VLB 1.5 mg/kg every 2 days for consecutive 5 times. The anti-tumor effects of different VLB preparations were compared,using living conditions,survival time,tumor volume and weight,and tissue pathological section as indexes. RE-SULTS:Compared with group A,B,C,D and E,the mice of group F were more active,and had longer survival time,smaller tumor volume and lighter tumor weight,with statistical significance(P<0.05). The tissue pathological section of mice in group F indicated that coagulation necrosis,disintegration,and dissolution of tumor cell nucleus. CONCLUSIONS:VLB hydrophilic group modified cationic liposomes have obvious anti-tumor effect,which are better than other VLB preparations.

Objective To analyze the amount and allocation of pediatric healthcare resources in Chengdu,and to recommend on local pediatric healthcare resources shortages.Methods Pediatric healthcare resources data of Chengdu came from pediatric relevant data reported regularly at yearend by the counties and districts to healthcare administration of the city.Data from such reports were subject to statistics with various indexes and descriptive analysis.Analyzed in focus was the distribution in 2015 of pediatric healthcare resources of the city among medical institutions of various types,levels and properties,as well as causes for such shortage.Results Tertiary hospitals hold 62.3% of the pediatric beds and 64.2% of pediatricians,and provide around 70% of the medical workload for pediatric outpatients and inpatients,upon the majority of pediatricians with master degree and above,and senior ones of/above associate chief physician titles;Tertiary hospitals have 58.7% of the pediatric beds and 49.7% of pediatricians,yet the outpatients served by specialized hospitals were 5% above tertiary hospitals.Conclusions The imbalance and shortage in total of pediatric resources in Chengdu result from stable source of manpower supply,high professional risk exposure,low income,and long training duration among others.Such measures as a better pediatrician development system,greater incentives for pediatric development,and enhanced development pediatric service consortiums,as well as greater support for private specialized pediatric hospitals.Those measures combined can effectively alleviate the shortage of pediatric resources.

Objective To study the protective effect of minocycline against ischemia-reperfusion injury after liver transplantation and its molecular mechanism after circulatory death in rats.Methods The rat donation after circulatory death (DCD) liver transplantation model was established by using magnetic-ring method.The donor and recipient were male SD rats.The rats were divided into sham operation group (SHAM group),liver transplantation group,minocycline group (MIN group),atractyloside + minocycline group (ATR + MIN group),24 rats in each group.In the MIN group,10 mg/kg minocycline was injected through the dorsum vein of the penis after reperfusion.The ATR + MIN group was injected with 2 mg/kg atractyloside.The open status of mitochondrial permeability transition pore (mPTP) was detected at 2,6,and 24 h after operation.Western blotting and immunohistochemistry were used to detect the expression of caspase3 and cyt c.Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were determined.Liver tissues were stained with hematoxylin-eosin (HE) and pathological changes were evaluated by Suzuki's standard.The survival of each group was calculated.Results As compared with liver transplantation group and ATR+ MIN group,the mPTP opening of MIN group decreased (P＜0.05),the expression of caspase3 and cyt c and the serum ALT and AST levels decreased significantly (P＜0.05),liver tissues injury was alleviated (P＜0.05),and the survival rate increased significantly after 30 days (P ＜0.05).There was no significant difference between liver transplantation group and ATR + MIN group (P ＞ 0.05).Conclusion Minocycline reduces ischemia-reperfusion injury in DCD liver transplantation in rats probably by inhibiting the mPTP opening,and preventing cyt c release and caspase3 activation to reduce hepatocyte apoptosis.This effect can be blocked by mPTP opener.

Objective To explore the clinical and pathologic features of Langerhans cell histiocytosis(LCH).Method Thirty-eight cases of LCH from both in hospital and outpatient department in dermatology and pediatry department from Jan.1991 to Jun.2006 were analyzed about the features of clinic,pathology and immunohistochemistry.Results The mean age of onset was 2.18 years old.The male /female ratio was 1.71.Skin lesions occurred in 78.9% of the patients.Among them,68.5% were as the first manifestation.The eruptions mainly distributed on trunk,90% of them presented as hemorrhagic maculopapules,nevertheless,3.3% of the eruptions showed as crust with hilar depression,which was similar to pityriasis lichenoides etvarioliformis acuta.Fever,hepatomegalia and splenomegia occurred in patients at a rate of 60.5%,68.4%,55.3% respectively.Thirty-one point six percent of the patients had got lymphadenectasis,the neck and inguinal lymph nodes were the common site to be affected.Ossature involvement occurred in 31.6% of the patients,which 8.3% got multiple injuries,howe-ver,91.7% got a solitary bone involved.Skull was the main site to be injured,else were lumbar,humerus,hipbone,an so on.Respiratory tract,auditory canal,mucosa were also the sites involved in this disease,but the incidence rate was lower than 10%,respectively.The laboratory data showed that 81.3% of the patients were anaemia,60.7% with abnormal subgroup of T-cell,and 32.1% positive for EBV-IgG.The skin histopathology data of 26/30 cases revealed that lichenoid infiltrates of Langerhans cells confined to the upper dermis.Cytologic features were cells with abundant eosinophilic cytoplasms,and longitudinally grooved or reniform nuclei.Lymphocytes and a few eosinophils also could be seen.Four cases of thirty showed that the proliferative Langerhans cells were with pale cytoplasms,besides,there were numerous eosinophils,and sometimes a few multinucleate cells were scattered.The immunity test of 20 cases of thirty displayed that CD1a(+)S100(+)KP-1(-).Biopsy of lymphaden and tumor of the skull of the rest 8 patients were all diagnosed as eosinophilic granuloma through both hematoxylin and eosin-stained section and immune marks.Conclusions Multiple systems can be involved in LCH.Hemorrhagic maculopapules,fever and splenohepatomegalia are common presentations in this disease.The morphous of nucles of histiocytes is particular,and to diagnose definitely,both CD1a(+) and S-100(+) are needed.

Objective To assess the safety, tolerability and efficacy of preoperative low dose intravenous amiodarone in the prevention of atrial fibrillation (AF) after off-pump coronary artery bypass grafting (OPCAB). Methods Two hundred patients with coronary atheroselerotic heart disease underwent selected OPCAB and were randomly divided into two groups: control group (100 cases) and experimental group (100 cases). Patients in control group were given conventional medicines and placebo, and patients in experimental group were treated with low dose intravenous amiodarone daily for 4 days before surgery. They were given conventional medicines after surgery. Results After the off-pump coronary artery bypass grafting, the incidence of AF, ventricular rate and the duration of AF in the experimental group were lower than those in control group [15% vs. 41%, χ2=16.766, P=0.000; (126.0±20.8) times/ min vs. (150.0±25.6) times/ min, t=0.478, P =0. 017; (8. 0±8. 6) h vs. (12.0±9.6) h, t=0. 439, P=0. 019]. No significant difference were found in the incidence of side effect between low close amiodarone group and the control group. Conclusions Prophylactic application of low dose intravenous arniodarone before off-pump coronary artery bypass grafting can safely and effectively reduce the postoperative ventricular rate and the incidence of AF, shorten the duration of AF, promote the rehabilitation and slightly improve the clinical symptoms.

Objective:To set up the HPLC fingerprints of Scutellaria baicalensis collected from different regions and develop a HPLC method for the quantitative analysis of the chief Flavonoids in Scutellaria baicalensis.Methods:The HPLC separation was performed on SHIMADZU VP-ODS column(250mm?4.6mm,5?m),gradient eluted using 0.1% citric acid-acetonitrile solvent system and detected at 276nm and 200nm to 400nm with a DAD detector.The flow rate was 1.0ml/min.The fingerprints of different samples were compared with similarity evaluation software.Results The HPLC fingerprints of Scutellaria baicalensis was set up,in the fingerprints,17common peaks of flavonoids were confi rmed.But the contents of the chief Flavonoids had a big difference,in which the baicalin had the biggest content(18.32%) in cottage Scutellaria baicalensis of Yiyuan County;the baicalein and wogonin had the biggest content(2.31%&0.73%) in wild Scutellaria baicalensis of Northeastern China.Conclusion:This method showed high precision,good repeatability,stability and all of the content were separated well,so it can be useful as the means to assess the quality of Scutellaria baicalensis.