Objective:To establish a detection method for residual toluene in diclofenac sodium raw material by GC. Methods:An HP-5 column (0. 53 mm × 30 m, 2. 65 μm) with nitrogen as the carrier gas was employed for the separation. The temperature of the column, injection port and FID detector was set at 50℃, 200℃ and 250℃, respectively. An internal standard method with isooctane as the internal standard was used to assay the residual amount after the sample was dissolved in DMSO. Results:The linear calibration curve was obtained over the range of 9. 27-111. 19μg·ml-1(r=0. 999 4). The mean recovery was 97. 27%(RSD=2. 80%,n=9). The LOD and LOQ was 0. 59μg·ml-1 and 1. 46μg·ml-1 , respectively. The residual contents of toluene in 3 batches of the products all met the relevant regulations. Conclusion:The established method is reliable and accurate, and can be used in the quality control of diclofenac sodium.

AIM: A methond was established for the determination of emodin and chrysophanol in Zhenhuang Powder (Margarita, Radix et Rhizoma Rhei, artificial Calculus Bovis, etc.). METHODS: RP HPLC was used for samples separated on a shim pack VP ODS column with the mobile phase of methanol-0.1% H 3PO 4(85∶15) and the detection wavelength was set at 430nm. RESULTS: The calibration curves were linear in the range of 0.01181～4.724?g( r =0.9994) for emodin and in the range of 0.01300～5.200?g( r =0.9999) for chrysophanol, respectively. The average recoveries were 99.0%( RSD =1.23%, n =5) and 100.5% ( RSD =1.06%, n =5), respectively. CONCLUSION: The method is simple, realizable and reproducible. It can be used effectively for the quality control of Zhenhuang Powder.

Objective:To observe the effects of Chinese herb baicalin on proliferation and cell cycle of periodontal ligament cells(PDLC) by stimulation of lipopolysaccharide(LPS) in vitro.Methods:Human PDLC was cultured in vitro.MTT colormetic method was used to evaluate the effects of Chinese herb baicalin on proliferation of PDLC by stimulation of LPS.Flow cytometry was used to analyze the possible DNA index and cellular cycle changes of PDLC.Results:PDLC showed a significant growth inhibition after being treaded with 100 ?g/ml LPS 3 h by MTT methods,and proliferation ability of PDLC improved greatly after adding baicalin.FCM results showed that the cell number increased in G1 phase,while the cell number decreased in S phase at 12 h after treated with 100 ?g/ml LPS,but the cell number decreased in G1 phase,and increased in S phase after treated with Chinese baicalin.Conclusion:Chinese herb baicalin can significantly promote the proliferation of PDLC.It can make some static cell in G1 phase enter S phrase and improve the growth of human PDLC.

Objective To investigate the influence of health care workers’(HCWs)hand hygiene intervention in healthcare-associated infection(HAI)in an intensive care unit(ICU).Methods Intervention measures of hand hygiene were adopted,effective supervision and management systems were established,hand hygiene compliance and HAI rate be-fore intervention(January-December 2012)and after intervention (the first stage:January-June 2013;the second stage:Ju-ly-December 2013)were compared.Results A total of 4 066 patients were surveyed,HCWs’hand hygiene compliance rate before intervention was 50.03%,the first and second stage after intervention was 61.80% and 64.57% respectively,there was increasing trend (rs=1.00,P<0.001).HAI before intervention was 5.48%,the first and second stage after inter-vention was 3.86% and 3.30% respectively,there was decreasing trend (rs=-1.00,P<0.001).Significant decreasing trends were found in rates of catheter-related bloodstream infection(CRBSI),catheter-associated urinary tract infection (CAUTI)and ventilator-associated pneumonia(VAP)(rs=-1.00,P<0.001).There were negative correlations between hand hygiene compliance rate and the infection rate of ICU,infection rate of CRBSI,CAUTI and VAP (P<0.05 ). Conclusion The improvement of hand hygiene compliance can effectively reduce the incidence of HAI in ICU ward.

Objective To measure the erythrocyte dielectric response variation of patients with KaschinBeck disease,to explore the pathogenesis of Kaschin-Beck disease and seek more effective means for evaluating the effectiveness of early diagnosis and control measures against Kaschin-Beck disease.Methods According to the principle of dielectrophoresis and cell dielectric response phenomena,dielectrophoretic pool was manufactured and dielectrophoretic detection and separation system was established.Patients with Kaschin-Beck disease (438) were selected in Kaschin-Beck disease areas of Aba County,Sichuan.At the same time,healthy persons (480) in KaschinBeck disease areas and non-Kaschin-Beck disease areas in Aba County were selected.Anticoagulant blood samples from patients with Kaschin-Beck disease and healthy persons were collected,erythrocytes were separated,cell suspension (10s cells/ml) was prepared and 200 μl cell suspension was added to the dielectrophoretic pool to measure erythrocyte dielectric response rate.Results Erythrocyte dielectric response rates were significantly different between patients living in Kaschin-Beck disease areas with Kaschin-Beck disease [(75.87 ± 5.89)％] and healthy persons living in Kaschin-Beck disease areas and non-Kaschin-Beck disease areas [(92.43 ± 4.45)％,(92.81 ±5.01)％,F =1.843,P ＜ 0.01).Erythrocyte dielectric response rate was significantly reduced in patients with Kaschin-Beck disease compared with that of healthy persons (all P ＜ 0.01);erythrocyte dielectric response rate of adult patients with Kaschin-Beck disease was lower [(69.57 ± 6.87)％] than that of pediatric patients with Kaschin-Beck disease [(82.17 ± 4.91)％,P ＜ 0.01].Erythrocyte dielectric response rates were significantly different in patients with different degrees of Kaschin-Beck disease (F =1.647,P ＜ 0.01).Erythrocyte dielectric response rate of patients with Kaschin-Beck disease was negatively correlated with prevalence of the disease (r =-0.87,P ＜ 0.01).Conclusions Erythrocytes of patients with Kaschin-Beck disease have some degree of pathological damage.The more severe the disease,the more serious the damage of red blood cell.The change of erythrocyte dielectric response properties may be used as an index to judge the prevalence of Kaschin-Beck disease and for early diagnosis.

Objective To understand the occurrence and risk factors of healthcare-associated infection(HAI) in a neonatal intensive care unit(NICU).Methods Neonates who were admitted to the NICU of a hospital from January 2012 to December 2014 were investigated retrospectively,risk factors for HAI were performed univariate analysis.Results A total of 760 neonates were included in the investigation,198 neonates developed 259 times of HAI,incidence of HAI was 26.05％,case incidence of HAI was 34.08％,incidence of HAI per 1 000 patient days was 9.50‰;the main infection site was lower respiratory tract (n =92,35.52％);among 259 cases of HAI,172 strains of pathogenic bacteria were isolated,the major pathogen was Acinetobacter spp.(n =40,23.26％);incidence of HAI was high in neonates with gestational age ≤32 weeks,birth weight≤1 500 g,length of hospital stay ≥10 days,duration of antimicrobial use≥10 days,mechanical ventilation,deep venous catheterization,and feeding intolerance,difference was statistically significant (all P＜0.001).Conclusion Incidence of HAI in NICU is high,effective prevention and control measures should be formulated according to its risk factors,so as to reduce the occurrence of HAI in neonates.

Objective To assess the relationship between the HLA gene sharing of couples and gestational diabetes mellitus (GDM). Methods Thirty GDM women and their husbands served as study group, 40 normal pregnant women and their husbands were selected as control group. All pregnant women were primipara with single fetus. 5 ml cubital venous blood were taken. Genomic DNA was extracted using phenol chloroform method. HLA-DRB alleles type were determined with PCR-SSP. Results The sharing of HLA-DRB1 between couples in GDM group were 33.33%, significantly higher than that of normal pregnancy(12.50%)(P0.05). Conclusions The sharing of HLA-DRB1 between couples may inerease the risk of GDM, It suggested that placenta- and-fetus unit may play roles in GDM.

Objective: Adopting quantified index to observe and analyze the effect of acute and chronic pain through double-wavelength diode laser. Methods: Using diode laser with the wavelength of 650nm / 810nm to irradiate the special tissues or aching points of patient, selecting different frequency, intensity and time to make treatment. Results: VAS scoring: P<0.05, the total efficient rate is 96%. Conclusions: Treatment with double-wavelength diode laser to various acute and chronic pain is just a definitely good curative effect, and it can release the effects of patient sleeping, daily life, communication with people and life interests caused by pain, and finally enhancing the quality of life.

Objective To study the antitumor activities and immunity regulation of Hedyotis diffusa Willd injection. Methods Inhibitory effects of Hedyotis diffusa Willd injection on cell proliferation was detected with MTT method. Then the inhibitive rate was calculated. The immunomdulatory effects of Hedyotis diffusa Willd injection were investigated. Results Hedyotis diffusa Willd injection inhibited the proliferation of A549, SGC-7901, HEP-G2, Helacells in the dose of 100～20 ?g/mL, and enhance the immunity regulation on S180 tumor transplanted in mice. Conclusion Hedyotis diffusa Willd injection has the antitumor activities and can enhance the immunity.

Objective:To efficiently amplify NK cells and determine their cytotoxic activity against a variety of tumor cell lines in vitro,thereby providing evidence for potential clinical application.Methods: PBMCs were isolated from adult peripheral blood and co-cultured with K562 cells that were genetically modified to express 4-1BBL,IL-15 and IL-21 on the surface for 15 days to effectively amplify NK cells.The total cell number and Purity of CD3-CD56+ cells were measured.Granzyme B and perforin expression of the amplified NK cells were detected by flow cytometry and real-time PCR.The anti-tumor effect on different cancer cells was evaluated.Results: This method obtained a more than 1.1×1010 CD3-CD56+ NK cells with 95% purity over a 15 day amplification procedure.The expanded NK cells could efficiently release granzyme B and Perforin.The cytotoxicity against different tumor cells was followed the order from strong to weak:gastric,pancreatic,cervical,ovarian and renal cancer cells,with the highest activity against gastric cancer cell line A549 (90% at E∶T=10∶1) (P<0.05).A time-dependent killing effect of activated NK cells on cervical,liver and pancreatic cancer cells was observed.Conclusion: This amplification procedure can consistently generate large amounts of pure NK cells with effective cytotoxic function against a variety of tumor cells.