Objective:To study the contents and determinate method of Sanggenon C of Cortex Mori in different sources.Methods:Sanggenon C was determined by HPLC. The mobile phase was methanol water(75∶25). Detection wavelength was at 280nm.Results:The average recovery was 96.94% with RSD = 1.72% and the linear range was 0.32～4.80?g. The highest content of Sanggenon C was 0.55% and the lowest content was 0.02% from different producing area. Conclusion: The method is accurate and reliable with good reproducibility.

Objective To study the impacts of B7-H3 molecule on the proliferation of T lymphocytes in different activation conditions and on the secretion of relevant cytokines.Methods Peripheral blood samples were collected from healthy subjects to separate peripheral blood mononuclear cells (PBMC) by Ficoll density gradient centrifugation.T lymphocytes were isolated from some of the PBMCs and purified with T Cell Enrichment Kit.PBMC and purified T lymphocytes were activated by anti-CD3/CD28 monoclonal antibodies (McAb) in vitro.Flow cytometry analysis was used to detect the expression of CD28 and CTLA-4 on T lymphocytes at different time points for further analyzing the activation states of T lymphocytes.On this basis, human B7-H3-Fc fusion protein was added into the mixed co-cultivation system on days 0, 1, 2, 3 and 4, and Hu-Fc fusion protein was used as isotype control.CCK-8 method was performed to detect the proliferation of T lymphocytes in each group.ELISA method was used to detect the secretion of cytokines (IL-2, IL-10 and IFN-γ) and to analyze the immune responses induced by stimulating T lymphocytes at different states of activation with B7-H3.Results B7-H3 molecule significantly inhibited the quiescent T lymphocytes from secreting IL-2 and IL-10, but had no significant impact on IFN-γ secretion.Moreover, it significantly promoted the activated T lymphocytes to secret IL-2 and IFN-γ, but had no obvious impact on IL-10 secretion.Results of the cell proliferation assay showed that B7-H3 molecule inhibited the in vitro proliferation of T lymphocytes in the PBMC, but had no obvious impact on purified T lymphocytes.ConclusionThe regulatory effects of B7-H3 molecule on the immune functions of T lymphocytes vary with the activation states of T lymphocytes.

Objective Verification of the serum proteomic fingerprints of NSCLC (non-small cell lung cancer) patients related to their advantage in Gefitinib therapy. Methods 27 NSCLC patients who have been treated with Gefitinib for more than 1 month and attained certain efficacy were taken for examination. They were divided into 3 groups, CR+PR, SD and PD, according to the evaluation criteria of solid tumor curative efficacy. All patients received SELDI (SELDI-TOF-MS surface-enhanced laser desorption/ionization time-of-flight mass spectrometry technique) inspection before treatment with Gefitinib. The shifting procedure was based on the abundance of the fingerprint :"M/Z 8693 50H+". According to the abundance of this fingerprint founded, the patients were divided into 4 populations: 1. The advantage population (10 patients, abundance 10 %);2. The disadvantage population (7 patients, abundance≥30 %);3. Suspected population A (5 patients, abundance 11%-15 %);4. Suspected population B (5 patients, abundance 16～ 29%). The ratio of patients in each of the 3 groups (CR + PR, SD and PD) into each of the above 4 populations were analyzed. Results It showed that: 1. In the advantage population, the ratio dispersed from CR + PR group is 9/10, those from SD group is 1/10 and from PD is 0.2. In the disadvantage population, the ratio dispersed from CR + PR group is 1/7, those from SD group is 1/7 and from PD is 5/7. In the suspected A group, the ratio from CR + PR group is 3/5, those from SD group is 1/5 and from PD is 1/5. 4. In the suspected B group, the ratio from CR + PR group is 3/5, those from SD group is 1/5 and from PD is 1/5. Conclusion This fingerprint "M/Z 8693 50H+" with abundance 10% after SELDI inspection is the index choosing advantage patients suitable to receive Gefitinib therapy.

Objective To investigate changes ane clinical significance of renal pathomorphology ane transforming growth factor-β1(TGF-β1 ),connective tissue growth factor(CTGF)of rats with eifferent stages of eiabetes mellitus(DM). Methods Thirty-six SD rats were raneomly eivieee into control ane DM group. Rats in DM group were given streptozotocin(STZ)at a eose of 65 mg/ kg boey weight ane rats in control group were given same amount citric acie/ soeium citrate buffer solution. Rats were killee at 4th,12th ane 24th weeks after DM moeels establishment. The renal hyeroxyproline concentration(HPC)was measuree. Masson′s trichrome was performee to observe perivascular collagen area(PVCA). Glomerular collagen eeposition score(GCDS),tubular interstitial eisease score(TIDS)in kieney interstitial tissue were measuree. Immunohistochemical staining were usee to measure the expression of type Ⅰ ane Ⅲ collagen,TGF-β1 ,CTGF. Renal path morphology change were observee with light microscope ane transmission electron microscope. Results (1)HPC levels of rats in DM group at 4th,12th,24th weeks were(216. 50 ± 10. 05)μg/ g,(267. 67 ± 11. 29)μg/ g,(340. 67 ± 39. 22)μg/ g,higher than that of control group((123. 00 ± 6. 00)μg/ g,(158. 50 ± 8. 89)μg/ g,(201. 00 ± 7. 69)μg/ g). The PVCA score in DM group were higher than that of control group( 4 weeks:(0. 45 ± 0. 07)vs (0. 96 ± 0. 11);12 weeks:(0. 60 ± 0. 06)vs(1. 65 ± 0. 18);24 weeks:(0. 72 ± 0. 07)vs(2. 63 ± 0. 0. 40);P< 0. 05). Meanwhile the scores of GCDS ane TILS in DM group)at 4th,12th,24th weeks increasee comparee with control group(4 weeks:((0. 34 ± 0. 03)vs(0. 14 ± 0. 01))× 103 / μm2 );12 weeks:((0. 49 ± 0. 03)vs (0. 26 ± 0. 04))× 103 / μm2;24 weeks:((0. 62 ± 0. 06)vs(0. 33 ± 0. 03))× 103 / μm2 )). The expression of CollagenⅠane Collagen Ⅲ in DM group were higher than that of control at 4,12,24 weeks after DM moeel establishment(Collagen Ⅰ:4 weeks:((4. 56 ± 0. 75)vs(1. 34 ± 0. 34))× 103 / μm2;12 weeks:((6. 07 ± 0. 74)vs(2. 17 ± 0. 12))× 103 / μm2;24 weeks:((7. 22 ± 0. 19)vs(3. 00 ± 0. 33))× 103 / μm2;CollagenⅢ:4 weeks:((3. 44 ± 1. 41)vs(1. 58 ± 0. 24))× 103 / μm2;12 weeks:((5. 60 ± 1. 34)vs(2. 95 ± 0. 41)) × 103 / μm2;24 weeks:((6. 22 ± 0. 18)vs(3. 27 ± 0. 27))× 103 / μm2 )),ane the eifferences between the groups all significant(P < 0. 05). Diffuse thickening of glomerular capillary basement membrane ane interstitial collagen hyperplasia in eiabetic rats were observee by electron microscope.(2)TGF-β1 expression in DM group were(2. 16 ± 0. 50)× 103 / μm2 ,(5. 31 ± 0. 25)× 103 / μm2 ,(6. 44 ± 0. 74)× 103 / μm2 respectively at 4th, 12th,24th weeks after DM moeel establishment,higher than that of control group((1. 18 ± 0. 15)× 103 / μm2 , (2. 99 ± 0. 25)× 103 / μm2 ,(3. 28 ± 0. 66)× 103 / μm2 )ane the eifferences were significant(P < 0. 05). The same trene was also seen in terms of CTGF expression((6. 02 ± 1. 39)vs(2. 32 ± 0. 87))× 103 / μm2 ,((6. 51 ± 0. 51)vs(3. 96 ± 0. 92))× 103 / μm2 ,((7. 89 ± 1. 46)vs(4. 58 ± 1. 02))× 103 / μm2;P < 0. 05) Conclusion Early eiabetic rats show renal fibrosis ane it may be relatee with overexpression of CTGF ane TGF-β1.

Objective To valuate the left ventricular function and myocardial fibrosis using Dopple tissue imaging(DTI) and integrated backscatter(IBS) in diabetic cardiomyopathy(DCM) rats. Methods Diabetes mellitus(DM) in healthy male SD rats was induced by single injection of streptozotocin (STZ,Sigma) into intraperitoneal at a dose of 65mg/kg body weight. Left ventricular fractional shortening (LVFS),ratio of peak early to late diastolic mitral inflow velocity(E/A) by color Doppler flow imaging-pulsed wave (CDFI-PW), ratio of peak early to late diastolic mitral annulus velocity (Ea/Aa) by DTI,myocardial calibrated IBS (IB%), cyclic variation of IBS(CVIB) by IBS, were determined in rats of 4,12 and 24 weeks after DM was induced. Heart weight index(HWI), Masson's trichrome staining, expression of connective tissue growth factor(CTGF) and activity of sarcoplasmic reticulum Ca2+ -ATPase(SERCA2a) in heart were measured, while the age-matched health rats served as control group. Results ① Diastolic dysfunction of left ventricular was found in diabetic rats detected by CDFI-PW(E/A<1) at the 4th week,and developed gradually. Pseudonormal filling(E/A>1) was found in major diabetic rats at the 24th week,which could be identified by DTI(Ea/Aa<1). The IB% of left ventricular posterior wall was significantly higher in diabetic rats than those in the control group( P<0.05), but the CVIB of the 24th week diabetic rats was lower(P<0.05). The LVFS had no great alteration in six groups(P>0.05).②Compared with the control group, the activity of SERCA2a was reduced, while HWI, collagen volume fraction(CVF),perivascular collagen area(PVCA) and the expression of connective tissue growth factor(CTGF) protein in heart were increased in diabetic rats (P<0.05), and correlation between IB% and HWI, CVF, PVCA,CTGF was significant (P<0.05). Conclusions DTI and IBS can assess early left ventrieular diastolic function and fibrosis of diabetic cardiomypathy in STZ-induced diabetic rats. DTI echoeardiography can identify "pseudonormal" mitral inflow patterns. DCM may be relatived with overexpression of CTGF and decreasion of SERCA2a activity.

After communication with professional course teachers, clinical experts and graduates by investigation, informal discussion and expert interviews, the course and teaching of rehabilitation therapy was reformed to make the students not only meet the skill requirement,but also acquire basic theory and sustainable develop in their career.

Objective To explore the relationship between left ventricular diastolic function and the dynamic changes of myocardial ultrastructure and argyrophilic fiber in diabetic rats on the different periods of lesions(week 4,12 and 24).Methods The diabetes mellitus(DM)in healthy male Sprague-Dawley rats was induced by a single injection of streptozotocin(STZ,Sigma)into intraperitoneal at a dose of 65 mg/kg body weight.The left ventricular diastolic function was measured by Color Doppler Flow Imaging-Pulsed Wave(CDFIPW)and Doppler Tissue Imaging(DTI)echocardiography.Heart tissue at the apex was obtained rapidly for transmission electron microscope study.Argyrophilic staining was used in the study of argyrophilic fiber volume fraction(APFVF)in heart interstitial tissue.Results Diastolic dysfunction of left ventricle was detected in STZinduced diabetic rats by CDFI-PW(E/A ＜ 1)at week 4,and progressed gradually.Pseudonormal filling (E/A ＞ 1)was found in diabetic rats at week 24,which could be identified by DTI(Ea/Aa ＜ 1).Diastolic function of normal rats was not impaired(E/A ＞ 1 and Ea/Aa ＞ 1).Transmission electron microscopy revealed a spectrum of subcellular remodeling in myocardium which was characterized by myofibril content decrease,disorganization,mitochondrial degeneration,sarcoplasmic reticulum,structural disorder.Compared with the control group,APFVF in myocardium was increased significantly in diabetic rats(P ＜ 0.05).Conclusion The diastolic dysfunction in STZ-induced diabetic rats correlates with damage of ultrastructure and increase of myocardial argyrophilic fiber.

Objective To discuss the clinical application of percutaneous transhepatic removal of gall-stone technique in treating common bile duct (CBD) stones.Methods Between January 2013 and January 2015,a total of 25 patients with CBD stones underwent lithotomy procedure via percutaneous transhepatic route.First,under ultrasound or fluoroscopy guidance percutaneous transhepatic cholangiography (PTC) was performed with subsequent placement of an 8 F rsheath;then,a balloon of 8-12 mm diameter was employed to dilate the papilla;mechanical lithotripsy was adopted when the stone size exceeded 12 mm;finally,through guide-wire exchange technique the stone-retrieval balloon was used to push the stones into the intestinal tract through the sphincter of duodenal papilla.Results The reasons to receive percutaneous transhepatic removal of gall-stone technique in the 25 patients included previous gastrointestinal surgery (n=18),endoscopic treatment failure (n=3),unwilling to receive endoscopic treatment (n=3),and other reasons (n=1).Successful removal of stones was accomplished in all 25 patients.After the treatment,complications occurred in 3 patients (12％),including fever (n=2) and liver abscess formation (n=1).The patients were followed up for 0.5-3 years;two patients died of tumor recurrence and metastasis,and one patient developed recurrence of common bile duct stones.No reflux cholangitis occurred.Conclusion For the treatment of CBD stones,percutaneous transhepatic removal of gall-stone technique carries higher technical success rate with lower incidence of complications,therefore,this technique can be used for the patients who are not suitable for endoscopic treatment or in whom endoscopic treatment failed.

BACKGROUNDTo establish a cohort for the study of risk factors of lung cancer, and to support the study of early biomarkers and prevention of lung cancer.

METHODSDesigned a special population-based prospectively dynamic cohort among radon- and arsenic-exposed tin miners aged 40 or more years old with at least 10 years of occupational exposure in Yunnan Province, P.R.China. The mass screenings with sputum cytology and chest X-ray were conducted annually. The baseline information was collected for assessing demographic characteristics and risk factors. The multiple sputum specimens, chest radiographs and numerous biologic specimens have been collected and stored.

RESULTSFrom 1992-1999, 9143 miners have been enrolled and 460 new cases of lung cancer have been found. There had 47655 person-time chest radiographs and 46625 person-time sputum cytology among the cohort in 8 years. The relative risks of age-adjusted exposure to chronic bronchitis, silicosis, and tobacco were 1.73, 1.46, and 1.32 respectively.

CONCLUSIONSA cohort of unique occupationally-exposed tin miners with an extensive biologic specimen repository and data bank has been successfully established. Although occupational exposures are the predominant risk factors among the high risk miners, lung cancer risk is also associated with chronic obstructive lung disease (chronic bronchitis and silicosis) and a number of measures of exposure to tobacco smoke, including early age of first use, duration, and cumulative exposure.

Objective To observe the expression of triggering receptor expressed on myeloid cells (TREM),Caspase-3 and interleukin (IL)-6 in optic nerve tissue of ischemic optic neuropathy (ION).Methods Twenty Sprague-Dawley rats were randomly divided into control group and model group,10 rats in each group.The permanent ligation of bilateral internal carotid arteries (BICA) was performed for 14 days to establish subacute ION model as model group.The control group were separated BICA without ligation.The expressions of TREM-1,TREM-2,Caspase-3 and IL-6 in rat retina were detected by reverse transcription PCR and Western blot,respectively.Results Compared with the control group,the expressions of TREM-1,Caspase-3,IL-6mRNA (t=6.058,7.86,6.055) and protein (t=9.671,9.524,14.501) in the optic nerve tissue of the model group were increased,while the expression ofTREM-2 mRNA and protein (t=9.283) was decreased,and the difference was statistically significant (P＜0.05).Conclusion In ischemic optic nerve tissue,TREM-1 mRNA and protein were significantly expressed,the expressions of TREM-2 mRNA and protein decreased significantly.