Taking the national key scientific and technological project in 11th Five-year and 10th Five-year Plan as its main clue,the innovative Chinese materia medica(CMM)research course has been reviewed in this paper.Based on the enlightenment from the first herbal medicine VeregenTM,which has been approved by FDA,together with the study and comprehension of many great achievements of new drug research,the strategy of innovative CMM research and development(R&D)and its practical route is going to be put forward in this article.With the innovative Compound Prescription of CMM research as the main innovative approach,innovation can be realized by the following three aspects:New substances,including new compound,new effective fraction/group or new components and their combinations;new indication of known compounds or extracts;the secondary development of high-quality Chinese patent medicine.Besides,in order to support the R & D of original drugs with Chinese characteristics,the international experiences in target-dependent screening and evaluation and optimization of leading compounds based on clinical application should be referenced,and the mechanism and targets of traditional Chinese medicine by genetic and molecular level be clarified.

0.05).The optimal combination was PCR-CTPP for MDR1 C3435T and PCR-SSP for G2677T/A.Conclusions PCR-CTPP and PCR-SSP are simple,accurate,rapid and economical methods for detection of SNP of MDR1 C3435T and G2677T/A,and can be applied in clinical research.

Objective To evaluate the role of acute kidney injury (AKI) in predicting the early (30-day) and late (30-day to 5-year) mortality of acute myocardial infarction (AMI) patients during hospitalization.Methods A total of 1371 adult patients diagnosed with AMI in the First People's Hospital of Changzhou from January 2008 to December 2012 were analyzed retrospectively with collecting their relevant clinical data from the hospital's database.AKI was categorized according to the 2012 KDIGO AKI criteria.To compare between death group and non-death group in AMI patients during 30-day and 30-day to 5-year.Different AKI stages of patients were compared,and their all-cause mortality were analyzed by Kaplan-Meier.Using multivariate COX regression analysis with two models to assess the factors for AMI patients in 30-day to 5-year.Results The prevalence of AKI after AMI in death group was higher than that in non-death group (the 30-day prevalence was 72.7％ vs 27.4％,P ＜ 0.001;the 5-year prevalence was 36.3％ vs 26.2％,P=0.013).In both early (30-day) and late (30-day to 5-year) follow up,the KDIGO grading distribution of AKI was different between death group and non-death group (P ＜ 0.001 in 30-day follow up and P=0.002 in 30-day to 5-year follow up).Among the 1371 AMI patients,410 (29.9％) developed AKI during the hospital stay.The 30-day and 30-day to 5-year mortality rates were 5.6％ (77/1371) and 11.3％ (146/1294) respectively.All-cause mortality and cardiovascular mortality were significantly higher in patients with AKI-Ⅰ stage,AKI-Ⅱ stage and AKI-Ⅲ stage than those with non-AKI (all P ＜ 0.001),especially in patients with AKI-Ⅲ stage.Further stroke history (HR=3.122,P=0.012),AKI severity (AKI-Ⅰ stage HR=3.034,P=0.028;AKI-Ⅱ stage HR=7.832,P＜0.001;AKI-Ⅲ stage HR=9.919,P＜0.001),and β-blocker therapy (HR=0.591,P=0.040) were independent predictors of 30-day mortality,while aging (HR=1.061,P ＜ 0.001),albumin (HR=0.943,P=0.023),AKI-Ⅲ stage (HR=3.944,P=0.007),β-blocker therapy (HR=0.660,P=0.041) and percutaneous coronary intervention (HR=0.256,P ＜ 0.001) were independent predictors of 30-day to 5-year mortality.Both at early (30-day) and late (30-day to 5-year) follow-up,AKI with or without baseline renal dysfunction were independent predictors of death in patients with AMI (all P ＜ 0.05).Conclusions AKI strongly correlated with short-and long-term allcause mortality of AMI patients,regardless of the baseline renal impairment.Specifically,the more severe AKI,the higher short-term mortality AMI patients have.

Objective To investigate the effect of albumin on expression of NLRP3 inflammasome and its downstream cytokines IL-1β and IL-18 in tubular epithelial cells.Methods Thirty mesangioproliferative glomerulonephritis (MsPGN) patients with different levels of proteinuria were selected, and their renal biopsy samples were stained by PAS and Masson to observe tubular epithelial cells injury and inflammatory cells infiltration.NLRP3, caspase-1, IL-1β and IL-18, as well as different inflammatory cells, were detected by immunohistostaining.In vitro, Western blotting and real-time PCR were employed to detect NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA in HK-2 cells stimulated by bovine serum albumin (BSA) (20 g/L).Results In MsPGN patients with high levels of proteinuria, there were obvious renal tubular epithelial cell injury and inflammatory cells infiltration (all P ＜ 0.05), and the expressions of NLRP3, caspase-1, IL-1β and IL-18 were up-regulated compared to patients with low levels of proteinuria (all P ＜ 0.05).Furthermore, IL-1β and IL-18expressions were positively correlated with the degree of proteinuria (r=0.836, P ＜ 0.05;r=0.901, P ＜0.05).NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA were significantly increased in HK-2cells stimulated by BSA compared to the control group (all P ＜ 0.05).Conclusions Albumin is able to induce NLRP3 inflammasome activation in tubular epithelial cells, which may be the mechanism of tubulointerstitial injury and inflammation caused by proteinuria.

Objective To establish a universal stem loop primer (USLP) based real-time PCR method to scan mature miR profile and quantify it's expression.Methods The common universal stem-loop primer pairs were re-designed; 8 random nucleotides were introduced at 3 ' end for reverse transcription of the mature miR,establishing a miR scanning and quantifying system based on SYBR Green Ⅰ PCR (improved USLP method).10-fold gradient diluted standard miRNA-155 cDNA ( 1 ～ 109 copies/μ1) were utilized to evaluate the sensitivity of this method.The specificity was verified by melting curve assay; the precision was assessed by intra-assay coefficient of variation (ICV) of threshold cycle (Ct value) through 20 repeated detections of the standard miR-155 cDNA (2 × 105,2 × 106,2 × 107 copies/μl) ; cost of the primers and time were evaluated,compared with that of the conventional USLP method.Peripheral blood samples were cultured with phytohaemagglutinin (PHA) for0 h,16 h,24 h,48 h and 72 h,and 87 candidate miR that may be associated with human immunity from PubMed data were scanned and quantified from the cultured T cells.Results The sensitivity of the improved USLP method was 103 copies/μl of standard miR-155 cDNA.Melting curve assay showed a single melting peak at 80 ℃,suggesting the excellent PCR specificity of miR-155.Precision of our method quantifying miR-155 was acceptable (ICV ＜ 2.5％ ).Compared with the traditional stem loop primers,our method saved 75％ cost of primers ( 1 917 bp vs 7 851 bp) and 60％ test time of reverse transcription (85 min vs 205 min).By our method,85 of the 87 miR expression in T cells had no significant difference after the PHA stimulation; the expression of miR-150 (72 h) decreased by 10 times and that of miR-155 (48 h) increased 8 times after culture with PHA (Z =-2.032,P =0.042;Z =- 2.023,P =0.043,respectively ).Conclusions The improved USLP method is fast,precise,sensitive,and cost-effective.It could be used for miR profile scanning and quantifying in T cells.

Objective To investigate the expression of monocyte-macrophage-related factors and interstitial fibrosis in kidney tissues of rats with ureter obstruction and recanalization .Methods Forty-eight male Spragur-Dawley rats were divided randomly into the obstructive group:sham (n=6), unilateral ureteral obstruction(UUO)3 days (n=6), UUO 7 days (n=6), and UUO 14 days (n=6) and recanalization group:bilateral ureteral obstruction(RBUO)0 day (n=6), 3 days after RBUO (n=6), 7 days after RBUO (n=6), and 14 days after RBUO (n=6).The kidneys were excised on day 3, 7, and 14, and the deposition of collagen fibers in kidney was detected with HE and Masson staining . Immunohistochemical analysis was performed to evaluate the protein expressions of monocyte chemoattractant protein -1 (MCP-1), macrophage colony-stimulating factor (M-CSF) and activated-macrophage marker CD68.Real-time PCR was used to detect the mRNA expressions of MCP-1 and M-CSF.TGF-β1 levels were determined by ELISA .Results Fibrosis observed with HE and Masson staining was obviously increased in kidney tissue of UUO rats , and aggravated as time prolonged, but alleviated in rats with recanalization .TGF-β1 levels were increased obviously in the UUO group , but decreased in rats with recanalization compared with those in BUO rats .In UUO rats, mRNA and protein expression levels of MCP-1 and M-CSF were increased .MCP-1 and M-CSF expression was gradually decreased in rats with recanalization compared with those in BUO rats .The dynamic change in expression of MCP-1 and M-CSF in both UUO rats and recanalization rats was consistent with the change in expression of CD 68. Conclusion Dynamic change in expression of MCP-1 and M-CSF in kidney tissues reflects change of activated and accumulated monocyte -macrophages , which may be one of the major mechanisms contributing to fibrosis induced by ureter obstruction .Renal fibrosis is alleviated by down-regulated expression of monocyte-macrophages factors with recanalization operation .

Aim To explore clinicalpathological features and the pathogenesis of lymphocytic mastitis, an uncommon breast benign disease. Methods The clinical pathological characteristics of 7 patients with lymphocytic mastitis were studied retrospectively. All cases were evaluated by using paraffin-embedded sections and by immunohistochemical staining with mouse anti-CD20, -CD45RO, and -CD68 antibodies. T and B lymphocytes infiltrated in the lobules of mammary gland were quantitatively analyzed according to stereoscope. The glucose regulation protein 94 (grp 94)/glycoprotein 96(gp 96), a member of heat shock protein family was also investigated in these cases by using immunohistochemical staining. Results It was showed that 4 cases were women suffering from insulin dependent diabetic mellitus (IDDM). One case was a woman without diabetic history. The history of the other two cases was not clear. The histopathological features of all 7 cases were lobulitis, perilobulitis and catheter ductitis with infiltration of lymphocytes accompanied with atrophy of lobule in mammary glands and homologous fibrosis of stroma. The result of immunohistochemical staining showed that most of infiltrated lymphocytes were B lymphocytes, while the small proportion of the cells were T lymphocytes, and the difference was significant(P〈 0.01). There was the expression of grp 94 in the cytoplasm of epithelium cells of lobules and ducts in normal mammary glands. A proportion of lymphocytes infiltrated in lobules and perilobules expressed grp 94. Some infiltrated cells expressed CD68. Conclusion A portion of lymphocytic mastitis is related closely to insulin dependent diabetic mellitus. Both humoral immunity and cellular immunity are probably involved in the pathogenesis of lymphocytic mastitis. Because of its unique pathologic and clinical features, lymphocytic mastitis should be defined as an independent mastitis and distinguished from other chronic inflammatory and fibrosing conditions in breast.