This article aims at exploring ways of integrating subject of pathobiology under the guidance of scientific outlook on development and improving and enriching pathobiology inte-gration and construction through curriculum systems recombination and integration,teaching con-tents optimization to reflect the latest research progress,network education platform establish-ment,comprehensive and exploratory laboratory course teaching system construction,new syl-labus and teaching material making up.

To fully incarnate"specialization"peculiarities of microbiology teaching courses by means of optimizing microbiology teaching contents and curriculum systems,and making up a new syllabus suitable to different specialties will lay a very solid foundation for cultivating medical talents with specialty traits.

Objective To study the clinical efficacy of intra-arterial thrombolysis with recombinant tissue plasminogen activator (rt-PA) for the treatment of ischemic cerebrovascular disease caused by cerebral thrombosis. Methods A total of 245 patients accepted by our hospital during May 2013 and July 2015 were divided into the observation group (n=148) and the control group (n=97). All patients were given conventional process for controling blood pressure and blood lipids. Patients in observation group received intra-arterial thrombolysis with rt-PA, while patients in control group accepted conventional treatment. At the time of admission, the demographic characteristic, vascular influencing factors, baseline clinical findings, laboratory findings and neurological deficits were collected. The improvement of neurological function was evaluated by the modified Rankin scale 3 months after treatment. The levels of fibrinogen (FIB), D-Dimer, activated partial thromboplastin time (APTT) and thrombin time (PT) were measured before and 24 h after the treatment. Results There were no significant differences in demographic characteristic and general clinical data between the two groups ( P>0.05). The proportion of patients with improved neurological function was significantly higher in observation group than that of the control group (83.11％vs. 53.61％, P<0.05). There were no significant difference in coagulation index and fibrinolysis index before treatment between the two groups (P>0.05). Twenty-four hours after the treatment, the levels of FIB, D-Dimer, APTT and PT were significantly improved in the observation group compared with those before treatment. The level of FIB was significantly decreased, D-Dimer was significantly increased, APTT and PT were significantly prolonged in observation group compared with those of control group (P<0.05). Conclusion The rt-PA can effectively dissolve thrombosis and correct the coagulation system and fibrinolytic system.

Pathogen biology is a main course of curricula in medical university.In developing quality courses,we optimized Pathogen biology course content,created the PBL teaching mode suitable for this course,set up network teaching platform,enhanced teachers' qualification,constructed comprehensive and exploratory experiment teaching system,wrote new teaching syllabus and other measures,made the Pathogen biology course construction more reasonable and perfect.

Objective; To clone, sequence and analyze the coding sequences of the Mr 15000 and Mr 9000 active segments of natural granulysin derived from human CTLs activated by allogenic antigen ,in order to establish the basis for further purifying and investigating the immune - impairing mechanism of granulysin. Methods; The coding sequences of the Mr 15000 and Mr 9000 granulysin gene were amplified from the total RNA of activated CTLs of healthy person after reverse transcription by Nested - PCR, inserted into pET32a ( + ) vectors and then transformed into E. Coli TOP10, respectively. The recons were identified by PCR, endonuclease digestion and sequencing. Results: The whole coding sequences of the Mr 15000 and Mr 9000 active segments were successfully cloned as expected. The accurate pET32a - Mr 15000 and pET32a - Mr 9000 recons were obtained through Colony - PCR, endonuclease digestion and sequencing. There lied polymorphism on the 119 th amino acid of the product encoded by GLS gene. Conclusion; The coding sequences of the Mr 15000 and Mr 9000 active segments of human granulysin can be obtained and cloned by the methods mentioned above and can be used for subsequent research.

Objective To explore the causes of subclinical hypothyrodism in children and the effects of the interventional therapy with thyroixine on the course of it.Methods Two hundreds children with subclinical hypothyroidism were measured for thyroglobulin antibody(TGAb),thyroid microsomal antibody(TMAb) in the blood serum,examined by colord Dopplor ultrasonic,examined by fine needle aspiraton cytology of the throid and measured the rate of 131I absorbed by thyroid in order to find out the causes of the disease.Two hundreds cases were randomly divided into two groups on the base of the cause of diseases,treatment group 100 cases and control group 100 cases.The treatment group were treated by throxine 25-75 ?g/d and the therapeutic dosage were chosen with the normal value of free triiodothyronine(FT3),free thyroxine(FT4)and high sensitive thyrotropin(sTSH) in the blood serum .After one year thyroxine therapy were stopped.Thyroid function was examined 6 months later after stopping the thyroxine.Results Among all of the causes of subclinical hypothyroidism in children,Hasgumoto′s thyroiditis accounts for 56%,simple goiter accounts for 26%,antithyroid drug accounts for 6%,the lack of thyroxine substitution therapy on the hypothyroidism accounts for 5% and undefined causes accounts for 7% .The thyroid function could keep normal for 1 year with an alternative therapy with thyroxine on subclinical hypothyroidism in children.Half a year later after stopping thyroxine,the thyroid function turned normal in most of the children.There were obvious differences in the ratio of cure and the ratio of effectiveness between treatment group and control group (t=20.2,3.2 Pa

Objective To investigate the effect of insulin on D_5 dopamine receptor expression and function in renal proximal tubule (RPT).Methods Immortalized RPT cells and D_5 receptor transfected HEK293 (HEK-D_5) cells were used in the study to investigate the effect of insulin on D_5 receptor expression and function,and those effects were compared in RPT cells from Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). The function of D_5 receptor was determined by measurement of the Na~+-K~+-ATPase activity in HEK-D_5 cells. Results Insulin increased D_5 receptor protein expression in a concentration and time-dependent manner in WKY RPT cells,but not in SHR.The basal level of D_5 receptor expression was higher in WKY cells than that in SHR cells. Stimulation with fenoldopam(D_1-like dopamine receptor agonist) inhibited the Na~+-K~+-ATPase activity;pretreat- ment with insulin increased the inhibitory effect of fenoldopam on Na~+-K~+-ATPase activity in HEK-D_5 cells. Conclusion The abnormal regulation of insulin on D_5 receptor expression and function might be involved in the path- ogenesis of essential hypertension.

Eukaryotic expression vectors carrying the small hairpin RNA (shRNA) for TRPC6 mRNA were constructed, and the effects of knocking-down TRPC6 on puromycin aminonucleoside (PAN)-induced apoptosis of mouse podocytes were observed. Two eukaryotic expression vectors containing small hairpin structure targeting TRPC6 named pGCsi-TRPC6A and pGCsi-TRPC6B were designed and synthesized. The plasmids were transfected into conditionally immortalized murine podocyte cell line by liposome. The changes in the TRPC6 mRNA and protein expression were observed by RT-PCR and Western blot after 48 h. Cultured podocytes were divided into four groups: control group, PAN treatment group, PAN treatment+shRNA transfection group, and PAN treatment+ negative control group. The expression of Bax and Bcl-2 mRNA and proteins was detected by RT-PCR and Western-blot respectively. The apoptotic rate of podocytes was measured by flow cytometry. The results showed that the expression of TRPC6 mRNA and protein was decreased in the podocytes when transfected with pGCsi-TRPC6A, and pGCsi-TRPC6B. The expression of Bax was increased, and that of Bcl-2 was decreased at protein and mRNA levels in the podocytes after treated with PAN for 48 h. These changes was attenuated by knocking-down TRPC6. Knocking-down TRPC6 could effectively decrease the PAN-induced apoptosis of podocytes. It was concluded that TRPC6 may play an important role in the PAN-induced apoptosis of podocytes. Knocking-down TRPC6 gene could effectively prevent the podocytes from apoptosis induced by PAN.

Objective To compare the safety and effectiveness between treatments with autologous platelet gel (APG) versus standard care for treating refractory diabetic dermal ulcers.Methods The 46 patients with proved nonhealing diabetic dermal ulcers were enrolled. Eligible for the study were patients with grade II/III ulcers according to Wagner, lasting for at least 2 weeks and with no signs of infection at recruitment.Patients were given their informed consent document and randomly assigned to two groups: standard care (ST, n=23) or standard care plus topic application of APG (APG, n=23) for twelve weeks.The treatment of blood glucose, blood pressure and lipids was optimized and the empiric antibiotic treatment was further adjusted according to the results of culture and sensitivity testing in all patients. APG treatment consisted of wound dressing with APG, followed by topical washing and cleaning. The APG was then covered with vaseline gauze for 72 hours, after which the ulcers were treated by standard care. Participants were seen thrice a week, twice a week, or at weekly intervals. Twelve weeks observation was set as the end point.Results The would of APG group were improved in 22 patients with ulcers healed completely and 1 case with would area reduced. In the ST group, 13 ulcers were healed, 6 worsened and 4 with would area reduced. The cumulative rate of ulcer healing was 95.7% in the APG group versus 56.5% in the ST group (P=0.002). The total effective rate in APG vs ST group was 100.0% vs 73.9% (P=0.009). By Kaplan-Meier analysis,the time-to-healing of ulcer and time-to-lutation of sinus were significantly different between two groups (log-rank, P=0.006, 0.000, respectively). No treatment-related adverse events were observed. Conclusions Treatment with APG in addition to standard care results in a significantly faster and better healing for a refractory diabetic dermal ulcer and does not raise any safety concerns. So APG treatment can be a valuable and effective aid in the management of diabetic foot ulcers.

Objective:To investigate the antigenicity of ClpC 2 and the feasibility of polyclonal antibodies of ClpC 2 as detected antibody.Methods:rClpC2 was induced with IPTG.The rClpC2 was identified by SDS-PAGE and Western blot ,and purified by affinity chromatography ,with which rabbit were immunized and the specificity of rabbit antiserum was detected by Western blot , the titer of rabbit antiserum against ClpC2 was detected by double immunodiffusion and indirect enzyme-linked immunosorbent assay (ELISA).The antigenicity of the rClpC2 was detected by ELISA.The polyclonal antibodies of ClpC 2 were prepared to detect the ClpC 2 in clinical serum of TB patients by ELISA.Results:SDS-PAGE showed specific protein band with a relative molecular mass of 46 kD.The rClpC2 could bind with the antibody in the blood serum of the mouse immuned by MTB.By Bandscan analysis rClpC 2 accounted for about 58.7%of the total bacteria protein ,the purity of rClpC2 was 88.5% after purification.The ClpC2 of BCG could bind with the rabbit antiserum.The titer of antiserum were 1∶32 and 1∶320 000 by double immunodiffusion and ELISA detected respectively.ELISA results showed that clinical serum positive rate of rClpC 2 antigen was 46%in TB patients,the sensitivity of this protein was 46%,and the spe-cificity of this protein was 90%.ELISA results showed that the sensitivity of rabbit antiserum against ClpC 2 was 40%, and the specificity was 90%.Conclusion: Successfully expressed and purified rClpC 2 and high titer polyclonal antibody were successfully prepared,and these results will provide basements for further study on the biological functions of ClpC 2 and its candidate potentiality as serological diagnosis and drug-target and biological functions of antiserum against ClpC 2.