OBJECTIVE:To provide reference for the instructions of domestic Clindamycin phosphate injection. METHODS:The contents in the instructions of domestic Clindamycin phosphate injection approved the listing from Jan. 2006 to Dec. 2015 were statistically investigated according to Regulation for Drug Package Inserts and Label Management and Regulatory Guidelines for Chemicals and Biologicals,and the items with great differences in different approval number were analyzed comparatively. RE-SULTS:The labeling rate of clinical experiment in the collected 25 instructions of domestic Clindamycin phosphate injection was 0,the labeling rate of validity was 44%,the labeling rate of use for elderly patients was 68%,the labeling rate of overdosage was 76%,and the labeling rate of others was 100%. The items with great differences were mainly special populations,usage and dos-age,adverse reactions,validity,storage conditions and precautions. CONCLUSIONS:There are great differences in instructions of domestic Clindamycin phosphate injection,some items are inaccurate,incomplete and lack of scientificity,which may result in ir-rational drug use in patients and increase the risk of medication. It is suggested that the domestic enterprises should modify and per-fect the contents of the instructions of domestic Clindamycin phosphate injections referring to the instructions of foreign brand name drug;and the drug regulatory department should strengthen the monitoring of drug instructions.

Objective To investigate the effect of MDR1 and MDR3 gene silence by shRNA of human breast carcinoma cell line MCF-7/Adr,and explore the role of MDR1 and MDR3 in adriamycin-resistance of breast carcinoma cells. Methods shRNA plasmid vector specifically targeting MDR1 and MDR3 gene was transfected into cells. The control group was transfected with empty vector. The concentration of adriamycin was detected by the flow cytometry (FCM). Cell apoptosis was analysed by FITC-Annexin-V/PI double staining. Cell viability and the IC50 of adriamycin on MCF-7/Adr cells were determined by MTT method. MDR1 and MDR3 mRNA were assessed by RT-PCR. P-gp expression was detectedby immunochemistry. Results After treatment with ABCB1 and ABCB4 shRNA plasmid vector, the apoptosis of MCF-7/Adr cells was (30.21±1.65)%and (22.07±2.17)% respectively. Compared with untransfecedgroup and empty vector transfection group the difference was significant(P＜0.01). MDR1 and MDR3 shRNAcould increase cellular adriamycin accumulation of MCF-7/Adr cells. MCF-7/Adr cells viability and the IC50were significantly decreased after transfection. Compared with untransfeced group and empty vector transfectiongroup, the mRNA level of MDR1 and MDR3 in MCF-7/Adr cells were decreased by (89.5±0.8)%and(85.1±1.2)%, the reduction of MDR1 and MDR3 mRNA was in a time-dependent manner. Immunochemistry proved that the expression of p-gp was significantly inhibited. Compared with untransfeced group and empty vector transfection group the difference was significant (P＜0.05). Conclusion The shRNA can effectively and specifically silence the expression of MDR1 and MDR3 gene, reverse the adriamycin-resistance mediated by P-gp in MCF-7/Adr cells. The reversal effect of adriamycin-resistance by shRNA of MDR1 is more effective than that of MDR3.

Animals ; Calmodulin ; metabolism ; Epimedium ; Flavonoids ; pharmacology ; Gene Expression ; Hypothalamo-Hypophyseal System ; drug effects ; metabolism ; Male ; Pituitary-Adrenal System ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To investigate the spatio-temporal expression of P2X3 receptor ( P2X3R) in rats with diabetic mechanical allodynia ( DMA ) .Methods DMA model in rats was induced by intraperitoneal injection of streptozocin ( STZ) .The von Frey filaments were applied to identify the changes of the paw withdrawal threshold ( PWT) in DMA rats.Immunofluorescence was employed to detect the spatio-temporal expression of P2X3R in the spinal dorsal horn (SDH), dorsal root ganglion (DRG) and hind paw skin on different time points after intraperitoneal injection of STZ , respectively.The protein expression of P2X3R in SDH and DRG was further semi-quantitatively analyzed by Western blotting.Results Compared with control group , intraperitoneal injection STZ induced significant mechanical allodynia indicated by the reduced PWT from 7 days, and which reached the peak on 14d and maintained to 28days (P<0.05). The expression of P2X3R in DRG neurons was significantly increased on 14 days and 21 days (P<0.05), while that in SDH and skin was markedly increased on 21 days and 28 days, compared with control group (P<0.05).Conclusion With the progress of DMA, the expression of P2X3R was significantly increased in the SDH, DRG and skin, which was almost parallel with the mechanical allodynia , but the changes in SDH and skin were 1 week later than that in DRG .These results suggest that P2X3R may play a key role in the maintenance of the DMA .

OBJECTIVE:To establish a RP-HPLC method for the determination of chlorogenic acid in Qingyan granules.METHODS:Waters Breeze System was used with a Hypersil BDS C18 column(5?m,4.6mm?150mm).The mobile phase was composed of methanol and 0.4%(V/V)H3PO4 water solution(20∶80,V/V),flow rate:1.0ml/min;wavelength of UV detector:327nm,column temperature 20℃.RESULTS:The calibration curve of chlorogenic acid was linear within the range of (174～1 392)ng(r=0.9 999).The average recovery was 100.10%,RSD=1.60%(n=5).CONCLUSION:This method is simple,sensitive,as well as reliable,and is available to quality control.

OBJECTIVE:To establish a method for the determination of dissolution of Xiaocaihu pill,and compare the differ-ence of preparation from different manufacturers. METHODS:Using 0.1 mol/L HCl as dissolution medium,rotating basket method was used to determine the dissolution of preparations. HPLC was adopted to determine the content of baicalin:column was TSKgel ODS C18 with mobile phase of methanol-water-phosphoric acid (65∶35∶0.7,V/V/V) at a flow rate of 1 ml/min,detection wave-length was 280 nm,column temperature was 30 ℃,and injection volume was 5 μl. RESULTS:The linear range of baicalin was 0.488-124.8 mg/L (r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2.0%;recovery was 100.14%-104.78%(RSD=1.58%,n=9). The average t50(50% dissolution time)of baicalin was 85.81 min. CONCLUSIONS:The method is simple with good precision,stability and reproducibility,and can be used for the dissolution determination of Xiaocaihu pill. Xiaocaihu pill from different manufacturers shows great differences,both preparation formulation and clinical use should attach importance to the dissolution of solid preparations.

Objective To evaluate the application value of 18 F‐FDG PET/CT in patients with fever of unknown origin (FUO) to improve the etiological diagnosis accuracy of PET/CT in patients with FUO and provide better service for clinical prac‐tice .Methods Clinical data and imaging results of 144 cases of FUO patients diagnosed through 18 F‐FDG PET/CT examination in Wuhan union hospital from December 2009 to December 2011 were retrospectively analyzed .The final diagnoses were based on sur‐gical pathology ,aspiration cytology ,or discharge diagnosis combined with more than 12 months of follow‐up results .All the PET/CT images were analyzed and the clinical application value of PET/CT in FUO patients were evaluated through the diagnose rate , diagnose efficiency and clinical staging of malignant tumor .Results Among the 144 patients ,there were 130 cases ultimately had clear diagnosis ,which including 41 cases of malignant tumor ,89 cases of benign lesions and 14 cases the cause of unknown .PET/CT detected abnormal lesions for 97 .6% (40/41) of patients with malignant lesions and 88 .7% (79/89) of patients with benign le‐sions ,and the diagnostic sensitivity ,specificity ,accuracy ,positive predictive value ,and negative predictive value were :97 .6% (40/41) ,86 .5% (77/89) ,90% (117/130) ,76 .9% (40/52) and 98 .7% (77/78) .PET/CT detected the primary lesions for the first time in 46 .3% (19/41) patients with malignant tumor ,and detected more lesions than the traditional imaging methods in 92 .7% (38/41) of these patients .Conclusion 18 F‐FDG PET/CT imaging can provided important reference value for the detection of primary lesions ,the qualitative diagnosis of benign and malignant lesions and clinical staging of malignant lesions .

Objective To verify the performance of quantitative detection of interleukin-6 by using IMMUNITE1000 chemilumi-nescence analyzer.Methods According to the requirements of International Organization for Standardization(ISO)1 5 189,serum specimen were collected and levels of IL-6 were detected.The precision,accuracy,analytical measurement range,reportable range amd normal reference range of quantitative detection of interleukin-6 by using IMMUNITE1000 chemiluminescence analyzer were verified,and its performance was evaluated.Results The coefficient variation(CV)of between-day precision of high and low value was 6.42% and 1.97% respectively,and that of within-run precision was 3.40% and 3.82% respectively.Compared the test re-sults with the target values,the bias % was 0.91%.The regression equation:Y =0.986X - 7.1 (r 2 = 0.999,P < 0.05 ).With 27 times diluted,the recovery rate was from 97% to 100%,and the clinical reportable range was 2 to 27 000 pg/mL.The 95% refer-ence interval ranged from 0 to 5.3 pg/mL.Conclusion The performance of this system meets the manufacturer′s declaration,and could satisfy the quality requirements of clinical laboratory.

Objective To analyze the epidemiological characteristics of Mycobacterium tuberculosis by enterbaeterial repetitive intergenic consensus-polymerase chain reaction(ERIC-PCR)DNA fingerprint. Methods Mycobacterium tuberculosis positive sputum samples between September 2003 to May 2006 were collected and cultured.Chromosomal DNA were extracted and ERIC-PCR DNA fingerprinting was analyzed by software,such as RAPD PHYLIP and Treeview.Results A total of 42 different fingerprints were detected.Phylogenetic analysis showed that they could be classified into three clusters,the clustering rate was 72.6%.The characteristics of ERIC-PCR fingerprint patterns were related to age,drug resistance,and type of resistance.Conclusions ERIC-PCR DNA fingerprinting technique used in this study is good for epidemiological studies with its strong discrimination,simplicity and rapidness.A high level of recent transmission is found in our city.

Objective To evaluate the changes in natural Killer (NK) cell subsets definetd by CD11b and CD27 in the spleen of septic mice.Methods A total of 168 pathogen-free healthy male C57BL/6 mice,weighing 20-30 g,aged 8-10 weeks,were divided into 2 groups (n=84 cach) using a random number table:sham operation group (Sham group) and sepsis group (Sep group).Sepsis was induced by cecal ligation and puncture in chloral hydrate-anesthetized mice.Thirty mice in each group were selected to assess the survival within 4 days after operation,and the survival rate was calculated.At 2,4,6,24,48 and 72 h after operation (T1-6),6 mice were randomly selected,and blood samples were taken from the eyeballs for determination of serum tumor necrosis factor-alpha and interleukin-10 concentrations by enzyme-linked immunosorbent assay.Six mice were randomly selected at T4-6 and sacrificed,and the spleens were removed for measurement of the percentage of NK cells and their subsets by flow cytometry.Results Compared with Sham group,the survival rate was significantly decreased at different time points,the serum necrosis factor-alpha concentration at T1-3,6 and serum interleukin-10 concentration at T3-6 were increased,the percentage of NK cells was decreased at T4-6,the percentage of CD27-CD11b+NK cells was decreased and the percentage of CD27+CD11b+ and CD27+CD11b-NK cells was increased at T5,the percentage of CD27-CD11b-NK ceils was decreased at T4,5,and the percentage of CD27-CD11b-NK cells was increased at T6 in Sep group (P＜0.05).Conclusion Changes in NK cell subsets defined by CD11b and CD27 in the spleen play an important role in the development of sepsis in mice.