The paper analyzed legally the following predicaments of legitimization of rural doctors in China:no legitimacy protection for their practice,vague criteria for practicing medicine,conflicts between legal regulations for medical practice and rural realities,and lack of a liability insurance system.The authors,based on an exploration of the legislative framework for rural doctors,made four corresponding legislative proposals as follows:to clearly define the legal scope and identity of rural doctors,to establish a qualification and employment system for rural doctors,to develop practicing norms applicable for rural doctors,and to set standards for their income.

Objective To investigate the effect of antioxidants including PDTC on pancreatic fibrosis of rats with chronic pancreatitis.Methods The rats were randomly divided into 5 groups including control group, CP group, PDTC treatment group, vitamin E treatment group and vitamin C treatment group.The CP model was in ducad by using intraperitoneal injection of DETC (750 mg/kg), twice a week.The control group received no treatment.After DETC injection, the treatment groups received an intraperitoneal injection of PDTC (100 mg/kg), vitamin E (15 mg/kg), vitamin C (15 mg/kg), respectively.Rats were sacrificed at 90 min, 24 h, 48 h, 72 h, 2 w, 3 w, 4 w, 6 w after first injection of DETC.Pancreatic tissue was taken for routine pathological examination.The activity of SOD, GSH-PX and MDA content were detected by spectrophotometric ratio method.α-SMA, desmin collagen Ⅰ, Ⅲ, TGF-β1, FN were detected by immunohistochemical assay.The expression of TGF-β1, FN mRNA was measured by RT-PCR.Results At 6w, the fibrosis and the parameters for damage of the pancreas in the three treatment groups were significantly better than that in CP group (P ＜0.01), the vacuolar degeneration index in vitamin E group and vitamin C group was also better than that in CP group (P ＜0.01).From the 2nd week, the activity of SOD, GSH PX in PDTC group, Vit C group and Vit E group was higher than that in CP group, while the MDA activity was lower than that in CP group, and the difference was statistically significant (P ＜ 0.01 or P ＜ 0.05).No significant difference was found among the three treatment groups.The mRNA levels of TGF-β1 and FN of the treatment groups were lower than those of CP group (P ＜0.05 or P ＜0.01), but higher than those of the control group (P ＜ 0.05).There was no significant difference among the three treatment groups (P ＞ 0.05).Conclusions PDTC and the other antioxidants can reduce oxygen free radicals by increasing the activity of SOD,suppressing the activation of PSCs, reducing the secretion of TGF-β1, Collagen Ⅰ , Ⅲ, FN and eventually inhibit the progress of pancreatic fibrosis.

Objective Based on the theory of preventive treatment and constitution regulation, to observe the clinical efficacy of point application for stable chronic obstructive pulmonary disease (COPD).Methods Seventy-nine qi deficiency or yang deficiency patients in stable COPD were randomized into a treatment group of 40 cases and a control group of 39 cases. The control group received conventional medication and the treatment group additionally received point application in canicular days or the three-nine days in winter. The frequency of catching cold, the situation of acute exacerbation of chronic obstructive pulmonary disease (AECOPD), constitution types and scores of traditional Chinese medicine (TCM), pulmonary function [the level of forced expiratory volume in one second (FEV1), the percentage of FEV1 in predicted value, FEV1’s ratio to forced vital capacity (FVC)], BODE index (body mass index, airflow obstruction, dyspnea and exercise capacity index) and therapeutic effects were recorded in the two groups before and after the treatment. The correlation analysis was performed between the scores of constitution and the frequency of catching cold, AECOPD, syndrome scores of TCM, BODE index.Results The cold times, AECOPD attack times, times of hospitalization for AECOPD and syndrome scores of TCM were significantly changed in the treatment group after treatment (P<0.01,P<0.05). There were statistically significant changes in the cold times, AECOPD attack times, and syndrome scores of TCM in the two groups after treatment (P<0.05). The total effective rate was 85.7% in the treatment group versus 70.3% in the control group, and the difference was statistically significant (P<0.05). The scores of the main abnormal constitution (qi deficiency or yang deficiency) and balanced constitution were significantly changed in the treatment group after treatment (P<0.01,P<0.05). There were statistically significant differences in the scores of the main abnormal constitution and balanced constitution between the two groups after treatment (P<0.01,P<0.05). There were no statistically significant changes in each item of pulmonary function in the two groups after treatment (P>0.05). After the treatment, the change of the BODE score in the treatment group was significantly different from that in the control group (P<0.01). A significant positive correlation was found between the main abnormal constitution scores and the cold times, AECOPD attack times, times of hospitalization for AECOPD, syndrome scores of TCM in the treatment group after treatment (P<0.05,P<0.01). A significant negative correlation was found between the balanced constitution scores and the cold times, AECOPD attack times, times of hospitalization for AECOPD, syndrome scores of TCM in the treatment group after treatment (P<0.05,P<0.01).Conclusion Point application is an effective method for stable COPD.

Objective To investigate the effects of interferon-α (IFN-α) and gefitinib on the proliferation and apoptosis of human colon cancer cell line HCT116. Methods Colon cancer cell line HCT116 was selected as research objective. The biological effects of IFN-α and gefitinib alone or combined on the cells were observed at different time point (after worked for 24, 48 and 72 hours). The proliferation inhibition of the medicine on the HCT116 cells was measured by methyl thiazolyl tetrazolium (MTT) assay. Morphologic changes were observed under optical microscope. Apoptosis was measured by flow cytometry (FCM). The results were analyzed with SPSS 13.0 software, two groups compare was tested by t test, and single factor variance analysis was for multiple group data compare. Results IFN-α and gefitinib alone or combined could significantly inhibit the proliferation of HCT116 cells (P<0.05), and there was a time and dose-dependent manner between the degree of inhibition and the working time and concentration of the medicine. With the work of the medicine, apoptosis morphologic changes were observed in the cells. And FCM result indicated that the apoptosis rate significantly increased. After treated with IFN-α and gefitinib alone or combined for 72h, the cell apoptosis rate were 15.6%±0.6%, 13.6%±0.4% and 31.2%±0.3% respectively, which was obviously higher than control group (6.8%±0.3%, P<0.05). Conclusion Both IFN-α and gefitinib were able to inhibit the proliferation and induce apoptosis of HCT116 cells moreover, and a synergistic effect was observed while combine used there two medicines.

Objective To investigate the pathologic changes in the pancreas of rats after intraperitoneal injection of DETC,a kind of superoxide dismutase (SOD) inhibitor,and to compared that with another model of chronic pancreatitis by pancreatic duct injection of TNBS.Methods The rats were randomly divided into DETC group,DETC control group,TNBS group,TNBS control group,normal control group.Rats in DETC group received an intra-peritoneal injection of DETC twice a week,and rats in DETC control group received an intra-peritoneal injection of same amount of normal saline.Rats in TNBS group was injected with 2％ TNBS ethanol phosphate buffer into the pancreatic duct,while rats in TNBS control group was treated with injection of same amount of ethanol phosphate buffer,and rats in normal control group received no treatment.The rats were sacrificed after 2 w,4 w,6 w and 8 w.The serum levels of amylase were determined,and pathological and ultrastructure changes of the pancreas were measured.The levels of SOD,GSH-PX activity and MDA content were detected.The expressions of α-SMA,Desmin,Collagen Ⅰ,Collagen Ⅲ,TGF-β1,FN in tissue were detected by immunohistochemical assay.The TGF-β1 mRNA expression was detected by RTPCR.Results No rat died in DETC group.The mortality rate of TNBS group was 15％.The serum levels of amylase were not statistically different between the 2 groups.The fibrosis scores of rat in DETC group at 4 w was 3.4 ± 1.l,which was significantly higher than that in TNBS group (3.0 ± 1.3,t =3.462,P ＜ 0.05).At 6 w,the damage scores of rat in DETC group was 9.1 ± 1.8,which was significantly higher than that in TNBS group (8.4 ± 1.8,t =2.943,P ＜ 0.05).Scores of vacuolar degeneration and fatty infiltration of rat in DETC group were higher than those in TNBS group,but the difference between the two groups was not statistically significant.Two weeks later,ultrastructure changes of pancreas could be observed,and large amounts of regenerative or mature collagen could be seen at 4 w.The SOD activity of DETC group was significantly decreased when compared with those in TNBS group (t =5.468,P ＜ 0.01).The GSH-PX activity of DETC group at 2 w,6w was significantly decreased when compared with those in TNBS group (t =6.497,10.125,P＜0.01).While the activity of MDA at 6 w,8 w was significantly increased when compared with those in TNBS group (t =3.350,5.407,P ＜0.05).The differences at other time points were not statistically significant.The expressions of (a)-SMA,Desmin,Collagen Ⅰ,Collagen Ⅲ,TGF-β1,FN,and TGF-β1 mRNA were not statistically significant between the 2 groups.Conclusions Sustained suppression of SOD activity can successfully induce chronic pancreatitis.Fatty infiltration and fibrosis in pancreas in DETC group occurs earlier with more severe presentation than that in TNBS group.Intraperitoneal injection of DETC is easy with low mortality rate,which is an ideal method for chronic pancreatitis model induction.

Objective To evaluate the diagnostic value of endoscopic ultrasound guided fine needle aspiration (EUS-FNA) in combination with flow cytometry (FCM) in lymphoma.Methods From January 2011 to December 2011,the cases of suspicious lymphoma with EUS-FNA examination at Shanghai Ruijin Hospital were retrospectively analyzed.The final diagnosis was according to pathological diagnosis of specimen from the surgery and follow up results.The sensitivity,specificity and accuracy of EUS-FNA combined with FCM in lymphoma diagnosis were initially analyzed.Results A total of 14 suspicious lymphoma patients were collected,eight cases were diagnosed as lymphoma by pathological examination of specimen from the surgery or.tissue from aspiration,four cases were non-lymphoma lesions and two cases still had no final diagnosis.The sensitivity and specificity of FCM alone in lymphoma diagnosis were 4/8 and 4/4 respectively.Six cases of lymphoma were detected by EUS-FNA with FCM.The sensitivity,specificity and accuracy of EUS FNA combined with FCM were 6/8,6/6 and 10/12 respectively.Conclusion EUS-FNA combined with FCM has better diagnostic value in lymphoma,especially for gastrointestinal lymphoma and those surrounding deep lesions.

Background Malignant osteopetrosis is an extremely rare dense bone disease,and sometimes features ocular disease and cranial nerve palsy.This disease received high attention because of its poor prognosis.And whether the eye manifestation improved after treatment is a problem for concern.Objective This study was to clarify the clinical manifestation,treatment and prognosis of malignant osteopetrosis associated with ocular disease.Methods A retrospective study was adopted.Two children with malignant osteopetrosis associated with eye symptoms were collected from Beijing Children Hospital.The systemic and ocular medical examinations were performed on the patients,including physical examination,hematology laboratory examination,abdominal B ultrasound and bone X ray radiography,external ocular examination,flash visual evoked potential (F-VEP) and CT of orbit.Bone marrow hematopoietic stem cell transplantation was employed and 5-year following-up was cinducted on the chidren.Results The children showed increased bone density,systemic bone sclerosis,basilar thickening,abnormalities of hematology indexes,anemia,hepatosplenomegaly,optical canal stenosis and abnormality of F-VEP P2 wave.In addition,optical disc pale,facial paralysis and paralytic esotropia were seen in a female child and alternating strabismus was found in another boy.After successful treatment,the systemic symptoms remitted in both children,but the eye findings remained unchanged in the female child during the follow-up duration.However,the strabismus diminished in the male child.The optical bone canal widening to 1.9 mm 1 year and 3.2 mm 5 years after treatment in the female child.Conclusions Strabismus and eye disease are the signs of malignant infantile osteopetrosis and reflections of the impairment of the central nervous system.The pathogenetic mechanism of malignant osteopetrosisrelated eye disease is below understanding now.Early bone marrow hematopoietic stem cell transplantation for malignant osteopetrosis can offer the best chance of long-term survival and improve the prognosis of eye diseases.

The aim of this project is to establish a fibroblast growth factor-21 (FGF-21) signaling pathway targeted cell model, for screening a class of FGF-21 receptor agonists as anti-diabetic candidates. FGF-21 requires beta klotho transmembrane proteins as co-receptor for the activation of tyrosine kinase FGF receptor (FGFR) signaling, thereby activating a series of intracellular signaling pathways and regulating gene transcription for glucose metabolism. Firstly a recombinant plasmid expressing co-receptor beta klotho and EGFP reporter genes was constructed. After introducing the recombinant plasmid into package cells, the cell culture supernatant was used to infect 3T3-L1 cells, which were then screened for stably expressing beta klotho gene. Administration of FGF-21 increased the expression of GLUT1 and stimulated GLUT1-mediated glucose uptake. This novel cell model can be conveniently used in high-throughput drug screening of FGF-21 or FGF-21 analogues.

Objective To construct and identify lentiviral vector pGC-FU-CXCR4 gene. Methods CXCR4 gene amplification was used by real-time polymerase chain reaction. The target gene fragments with the digested plasmids were exchange. Then the lentiviral vector pGC-FU-CXCR4 was constructed successfully. Use the constructed lentiviral vector to infect the competent escherichia coli cells. Polymerase chain reaction analysis was used to identify the cultural clones and DNA sequencing and comparative analysis were used to positive fragments. The successfully constructed plasmids had the same sequence with the target gene. Results Polymerase chain reaction tests showed that am-plified target genes were inserted in pGC-FU vectors. The electrophoresis results,digestion showed that the reconstructed plasmid was consist-ent with the theoretical fragment and the sequence result of the positive fragments were exactly the same with the target gene. Conclusion Lentiviral vectors of CXCR4 gene over-expression were successfully constructed.

of NB4 cells at S phase was lower than those in control group (P <0.05),and the number of NB4 cells at G1 phase was increased (P <0.05).The expression levels of Bcl-2 in ICA groups were significantly lower than that in control group (P < 0.05 ), while the expression levels of Bax were higher than that in control group (P < 0.05 ). Conclusion ICA can induce the apoptosis of NB4 cells via inhibiting the expression level of Bcl-2 and up-regulating the expression level of Bax.