Objective Construction of human IL‐4 recombinant expression vector and then conduct the expression ,purification and identification of human recombinant IL‐4 .Methods the open reading frame of IL‐4 was amplified by nest PCR with total RNA from PBMC of healthy volunteer .And then the amplified IL‐4 was inserted into pET101/D‐TOPO ,transformed into BL21 ,ex‐pressed ,purified and indentified .Results The size of amplified open reading frame of IL‐4 was about 460 bp and the sequence was correct .After transformed into BL21 ,the IL‐4 clone with higher expression level was selected by selection of different clones insert‐ed with IL‐4 and the size of expressed ,purified IL‐4 was about 28 × 103 .Western blot results showed that the size of single band was identical with the expected protein .Conclusion Human IL‐4 recombinant protein was got successfully .

AIM: In order to search carrier material and better tissue culture method, the morphology and structure of the cultured cat corneal endothelium was observed. METHODS: The cat corneal endothelial cells were seeded on the Descemet's membrane of the dehydrated swine corneal stroma, and then cultured in the medium with epidermal growth factor and laminin for 7 days. The morphology and structure of reconstructed tissue was tested by the inverted microscope and the scanning electron microscope. RESULTS: As shown by the morphological observations, the cultured cat corneal endothelial cells formed an integrated membrane, and attached to the Descemet's membrane, which was similar to the nature tissue. The cells connected tightly to each other, and some of them arranged in hexagon approximately. CONCLUSION: The cat corneal endothelial cells could be rebuilt on the carrier of the dehydrated swine corneal stroma successfully, which is similar to the nature tissue.

0.05). (2) In the total 12 rabbits, all the cornea grafts survived without rejection reaction, corneal haze or corneal neovascularization. The fresh grafts got transparent after 2 months, and the dehydrated grafts got transparent after 6 months. Histological study demonstrated both fresh and dehydrated stroma grafts had fused with rabbits'corneal stroma very well without lymphocytes infiltrating. (3) As shown in histological observations, the reconstructed cat corneal endothelial tissue was similar to the nature tissue. Cultured cat endothelial cells connected tightly to each other and attached to the Descemet's membrane closely. CONCLUSION: Swine corneal stroma has low immunogenicity and satisfying biocompatibility,it is an ideal biological carrier for cornea reconstruction in vitro.

Objective:Expression and purification of the α-HL of Staphylococcus aureus as antigen for making full human anti-α-HL antibody later ,providing of new treatment for Staphylococcus aureus infection.Methods:The total RNA of Staphylococcus aureus was extracted and the cDNA of α-HL was amplified by RT-PCR.The DNA of α-HL and pCold-TF plasmid was digested and ligated by T4 ligase and then transformed into E.coli TOPO 10.The recombinant plasmid α-HL/pCold-TF which verified by sequencing was trans-formed into E.coli BL21 for expression.The expression products was identified by SDS-PAGE and Western blot.Results: The size of amplified cDNA of α-HL was about 900 bp and the expressed soluble fusion protein of α-HL was about 90 kD(including the molecular chaperone in the vector ) after inducing expression for 24 h at 15℃.The Western blot results showed that the expressed protein was the fusion protein of α-HL.The purified α-HL was injected into BABL/c mice for making antiserum.The results showed that the antiserum had good binding activity with Staphylococcus aureus and the titer was greater than 10 000 times.Conclusion: The α-HL of Staphylococcus aureus was successfully cloned and the soluble fusion protein of α-HL was successfully expressed.

Objective:To develop non-immunized human phage display library.Methods:The total RNA of lymphocyte cells from peripheral blood of healthy voluntee was isolated and cDNA was synthesized,and the genes of heavy variable chain (VH) and light variable chain (V? and V?) were amplified by direct PCR and half-nested PCR.By overlapping extension PCR,the genes of VH and VL (V? and V?) were linked.The linked genes of single chain Fv fragment (scFv) were ligated with the vector pCANTAB-5E and then cloned into TG1 for the scFv library construction.Results:By direct PCR and half-nested PCR,42 VH fragments,16 V? and 18 V? fragments were obtained.The size of linked scFv library genes was 750 bp and the volume of constructed scFv library was 1.35?108.The results of BstN Ⅰ analysis of scFv genes from the phage library showed that fingerprint map of the selected scFvs was different.Conclusion:The developed phage library is diversity and can be used for selecting humanized scFv.

Objective:To observe the clinical effect of tuina plus foot bath with Chinese medicine for patients with diabetic foot (DF) in early stage.Methods:A total of 70 patients with early-stage DF were randomly allocated by the random number table into two groups,with 35 cases in each group.Patients in the control group received conventional medication,while patients in the observation group received tuina plus foot bath with Chinese medicine on the basis of conventional medication.The clinical efficacy was compared after 2 courses of treatment.Results:After treatment,intra-group comparisons of ankle-brachial index (ABI) showed statistical significance in both groups (both P＜0.05).The curative rate was 83.3％ in the observation group,with the total effective rate of 96.7％,versus 29.4％ and 76.5％ in the control group,respectively,and the between-group comparisons showed statistical significance (both P＜0.05),indicating a better effect in the observation group.Conclusion:Tuina plus foot bath with Chinese medicine has a good therapeutic effect for DF patients in early stage.

Objective To evaluate the diagnostic value of endoscopic ultrasonography (EUS), and explore the efficacy of endoscopic treatment in patients with esophageal submucosal tumor. Methods Sixty-eight patients with esophageal submucosal tumor were selected, and the tumor was derived from the muscularis mucosa and submucosa according to the common endoscope and endoscopic ultrasonography detection. Endoscopic mucosal resection (EMR) was applied to remove submucosal tumor with diameter less than 1.0 cm, endoscopic piecemeal mucosal resection (EPMR) or endoscopic submucosal dissection (ESD) was applied to remove submucosal tumor with diameter 1.1 - 1.5 cm, and ESD was applied to remove submucosal tumor bigger than 1.5 cm. Samples were examined by pathology after treatment. Results Tumors in all the patients were completely removed, and the tumor diameter was 0.6-2.3 cm. Forty-one cases were treated with EMR, 9 cases were treated with EPMR and 18 cases were treated with ESD. Four patients had intra-operative bleeding that was stopped by electrocoagulation hemostasis. No perforation occurred in all cases. Postoperative pathology revealed 43 cases had leiomyoma, 23 cases had interstitialoma, and 2 cases had lipoma. Patients were reviewed by gastroscope 3 months after operation. The white scars formed in all patients, and there was no residue or recurrence. Conclusions Different origin layers and property of esophageal submucosal tumor can be diagnosed accurately by EUS, and endoscopic therapy (EMR, EPMR and ESD) is an effective treatment for submucosal tumor from muscularis mucosa and submucosa. Endoscopic therapy is safe and effective. It provides sufficient pathological information.

Objective:To review and analyze the prescriptions for Chinese herbal pieces in an integrative traditional Chinese and western medicine hospital to explore the trend and problems in clinical application for Chinese herbal medicines and provide reference for dispensing in pharmacy of traditional medicine and rational drug use. Methods: Totally 1200 prescriptions for Chinese herbal pieces in our hospital were surveyed from January to December in 2014. The irrational use was analyzed, and reasonable suggestions were provided for the clinical application. Results:The problems in the prescriptions for Chinese herbal pieces included uncompleted diagnosis,incompatible diagnosis and medication,nonstandard Chinese name,unclear footnote,irrational usage or dosage and incompati-bility. The prescriptions including above 15 varieties of Chinese herbal pieces in one accounted for 65. 70%, and those with average dose above 250 g accounted for 40. 25%. Conclusion:Regular inspection and quality supervision on the prescription for Chinese herb-al pieces should be strengthened to improve the quality of prescriptions and guarantee the safety and economy of the medication.

BACKGROUNDThe self-consciousness and practicality of preferentially prescribed essential medicines (EMs) are not high enough in county hospitals. The purposes of this study were to use the information-motivation-behavioral skills (IMB) model to identify the predictors of essential medicines prescribing behavior (EMPB) among doctors and to examine the association between demographic variables, IMB, and EMPB.

METHODSA cross-sectional study was carried out to assess predictive relationships among demographic variables and IMB model variables using an anonymous questionnaire administered in nine county hospitals of Anhui province. A structural equation model was constructed for the IMB model to test the instruments using analysis of moment structures 17.0.

RESULTSA total of 732 participants completed the survey. The average age of the participants was 37.7 ± 8.9 years old (range: 22-67 years old). The correct rate of information was 90.64%. The average scores of the motivation and behavioral skills were 45.46 ± 7.34 (hundred mark system: 75.77) and 19.92 ± 3.44 (hundred mark system: 79.68), respectively. Approximately half (50.8%) of respondents reported that the proportion of EM prescription was below 60%. The final revised model indicated a good fit to the data (χ2 /df = 4.146, goodness of fit index = 0.948, comparative fit index = 0.938, root mean square error of approximation = 0.066). More work experience (β = 0.153, P < 0.001) and behavioral skills (β = 0.449, P < 0.001) predicted more EMPB. Higher income predicted less information (β = -0.197, P < 0.001) and motivation (β = -0.204, P < 0.001). Behavioral skills were positively predicted by information (β = 0.135, P < 0.001) and motivation (β = 0.742, P < 0.001).

CONCLUSIONThe present study predicted some factors of EMPB, and specified the relationships among the model variables. The utilization rate of EM was not high enough. Motivation and behavior skills were crucial factors affecting EMPB. The influence of demographic variables, such as income and work experience, on EMPB should be fully appreciated. Comprehensive intervention measures should be implemented from multiple perspectives.

Adult ; Aged ; China ; Cross-Sectional Studies ; Female ; Hospitals, County ; statistics & numerical data ; Humans ; Male ; Middle Aged ; Practice Patterns, Physicians' ; statistics & numerical data

Sturge-Weber syndrome (SWS) is a neurocutaneous syndrome, characterized by the association of facial port-wine hemangiomas in the trigeminal nerve distribution area, with vascular malformation(s) of the brain (leptomeningeal angioma) with or without glaucoma. Herein, we reported Sturge-Weber syndrome in a 50-year-old man, who presented port-wine hemangiomas and epilepsy. In this case, the patient's epilepsy episodes from his first year of life had been ignored and separated from the entity of SWS by his physicians, which led to delayed treatment. This case illustrates the importance of careful examination of patients of any age with hemangiomas in the trigeminal nerve with concomitant episodes of epilepsy. In such cases, there should be yearly neuroimaging screenings to guaranteed early interdisciplinary interventions from the time of definite diagnosis.
Brain ; Epilepsy ; Glaucoma ; Hemangioma ; Humans ; Mass Screening ; Middle Aged ; Neurocutaneous Syndromes ; Neuroimaging ; Sturge-Weber Syndrome ; Trigeminal Nerve