OBJECTIVE: To establish RP-HPLC method for the determination of fluconazole in human plasma.METHODS: After liquid-liquid extraction,the plasma sample was analyzed on Diamonsil C18 column with column temperature set at room temperature.The mobile phrase consisted of methanol-0.07% aqueous solution of triethylamine(50∶ 50) with a flow rate of 1mL? min-1.The UV detection wavelength was set at 210nm,and phenacetin was used as the internal standard.RESULTS: The linear range of fluconazole was 0.15～ 10.0mg? L-1(r=0.999 2),with lower quantification limit at 0.15mg? L-1.The intra-day RSD was between 3.37% and 7.48% and the inter-day RSD was between 7.03% and 12.05%;the method recovery was 103.64%(RSD=5.04%),and the average extraction recovery was between 83.81%(RSD=5.77%).CONCLUSION: This method was sensitive,accurate,rapid and reproducible,and suitable for the content determination of fluconazole in plasma.

Objective To develop a novel approach of transfecting the exogenous gene safely and effectively mediated by receptor according to the cavitation effect of ultrasound-mediated microbubble destruction. Methods C-myc antisense oligonucleotide(ASODN) and galactosylation polylysine(G-PLL) which was fluoresce in-labeled specificity ligand and SonoVue microbbule were coupled by electrostatic interaction. SMMC-7721 cell with c-myc ASODN were transfected in vitro by ultrasound-mediated microbubble destruction to investigate the effect of c-myc ASODN on it. The c-myc mRNA gene expression of liver cancer SMMC-7721 cells was detected. Results Cultured with 10% SonoVue microbubble and under ultrasound at 1.5 MHz,1.0 W/cm2, 10% duty cycle,60 s respectively,the SMMC-7721 cells had not been changed significantly. Reverse transcription polymerase chain reaction(RT-PCR) assay showed that c-myc expression was decreased in transfected cells(P<0.01). Conclusions Receptor-mediated targeted microbubble destruction by ultrasound can enhance the exogenous gene transfection and expression. It is a promising method in gene therapy of liver cancer.

Objective MiRNA-10b is an important member of the MiRNA family , which has been proven that miRNA-10b can promote the growth and invasion of a variety of tumor cells .The aim of this study was to to investigate the effect and mechanism of miR-NA-10b on proliferation and invasion of low metastasis of lung cancer cell line (95-C). Methods The recombinant of miRNA-10b was transfected into 95-C by lipofectin method .The experiment set up 3 groups:blank control group , negative control group and miRNA-10b expression plasmid transfected group .MiRNA-10b expression level and KLF4mRNA expression level were detected by real-time fluores-cence quantitative PCR ( RTFQ-PCR) .The cell proliferation was detected by cell proliferation assay .The invasive ability of cell was de-tected by Transwell experiment .The expression of KLF4 protein was assessed by Western blot . Results At 48 hours after transfection, compared with blank control group (1.01 ±0.08) and negative control group (0.86 ±0.07), the miRNA-10b expression level in miRNA-10b expression plasmid transfected group (1.61 ±0.12) increased significantly (P<0.05) and there was no statistical difference between blank control group and negative control group (P>0.05).From the growth curve, the cell proliferation rate was obviously increased in miRNA-10b expression plasmid transfected group ([188.0 ±15.1]/HP) compared with the other two groups ([151.0 ±11.3]/HP), ([136.0 ±10.8]/HP) (P <0.05) and there was no statistical difference between these two groups ( P >0.05 ).Transwell result showed more cells transferred to the other side of Transwell compared with the other two groups ( P <0.05 ) and there was no statistical difference between these two groups (P >0.05).The expression of KLF4 protein decreased in miRNA-10b expression plasmid transfected group compared with the other two groups ( P<0.05).KLF4mRNA expression decreased, but the difference had no statistical significance (P>0.05). Conclusion MiRNA-10b might promote the pro-liferation and invasion of 95-C through down-regulation of KLF4 protein expression .

Objective To investigate the urinary albumin excretion of the diabetes patients and application value in the monitoring of early impairment in kidney. Methods The random urine samples from diabetes patients and controls within three days were collectod. The changes of urinary albumin excretion within day and between days were analyzed. 24-hour urine albumin was used as a standard to evaluate early kidney damage. The correlations between results of random urine albumin at the different time points and different periods were comparod. The sensitivity and specificity of random urine albumin at the different time points and different periods was evaluated and compared to deduce the best diagnostic porformance of the random urine albumin. Results There are greater variations of the levels of urinary albumin of patients with diabetes and control. After the correction with urine creatinine and urine volume the variations can be reduced (CV:49%±23% and 64%±30%). Urinary albumin excretion rate change rhythmically within the 24 h in healthy and diabetes patients. We found the best correlation between overnight ratio of urinary concentrations of albumin and creatinine (ACR) and 24-hour urinary albumin (R2 = 0.976). It was superior to urina sanguinis (R~2 = 0.900), postprandial urine (R~2 = 0.584) and random urine (R~2 =0.791). When 24 h urinary albumin was taken as the standard, receiver operating characteristic (ROC) curve analysis showed there was significant difference between male and female(male 12.8 μg/mg urine creatinine vs female 27.0 μg/mg urine creatinine),and the the cut-off value of ACR was 27.7μg/mg urine creatinine. When the smallest available negative likelihood ratio (0.011) and the greatest positive likelihood ratio (481.000) were obtained,the concentration of 13.0 μg/mg creatinine and 87.4 μg/mg creatinine were set as the cut-off value of ACR. Conclusions The correction with urinary creatinine can reduce the variation between-days compared with urine volume, but still can not completely eliminate the variability. The ACR of overnight urine has the best correlation with the 24 h urinary albumin and can replace 24 h urinary albumin. Random urine as the most convenient collecting urine samples can also replace 24-hour urinary albumin, but the gender discrepancy need to be considered. When the concentration of 13.0 μg/mg and 87.4 μg/mg was set as a random ACR exclusion value and the confirmative value, it can basically rule out and confirm the existence of microalbuminuria.

Urinary albumin used as an important indicator in diagnosis of renal disease , but there are still many problems in the clinical application .The urine specimen type , threshold value , reference measurement procedures and some more aspect were summarized in this article .

Objective To study the efficacy of medication management, symptom management and community reentry skills-training modules to prevent rural patients with schizophrenia from relapse and increase their social function.Methods 89 subjects were randomly assigned to the skills training group (45 cases) and the control group (44 cases). Both groups received the same treatments, but the skills training courses were given to the skills training group for twelve weeks when the subjects were addmitted into the trial and at their one-year follow-up timepoint respectively. And a two-year follow-up was carried out. All subjects were evaluated with PANSS, SDSS, relapse and employeement.Results 89 subjects had completed the study. The skills training group demonstrated clinical Results significantly superior to those of the control group on overall improvement according to PANSS and SDSS. Substantially, statistically significant advantages of the skills training group were related to 1) the rate of relapse (11.1% vs 31.8%), 2) the rate of employment (37.8% vs 15.9%).Conclusion Medication management, symptom management and community-re-entry skills-training modules are effective in reducing patients' psychiatric symptoms, preventing them from relapse and increasing their social function.

Objective:To study the effects of miR-20a on the osteogenic differentiation potential of inflammatory periodontal liga-ment cells(IPDLSCs).Methods:Cells were isolated and cultured from the healthy and inflammatory periodontal ligament samples (HPDLSCs and IPDLSCs)respectively.miR-20a expression was analyzed by qRT-PCR.Alizarin red staining,Western blot and PCR were used to evaluate the osteogenic differentiation potential of IPDLSCs after transient transinfection of miR-20a mimics or inhib-itor.Results:miR-20a expression in IPDLSCs was lower than that in HPDLSCs,and the osteogenic differentiation potential of IP-DLSCs were promoted by miR-20a mimics,and reduced by miR-20a inhibitor.Conclusion:The miR-20a in IPDLSCs was down reg-ulated.miR-20a can promote the osteogenic differentiation potential of IPDLSCs.

We report a case of a 44-year-old female patient with acute liver failure and hepatic encephalopathy. The patient received artificial liver treatment and underwent allogeneic orthotopic liver transplantation at 14 days after admission. Laboratory examination reported a small number of green cytoplasmic inclusions in neutrophils on the peripheral blood smear at 68 days after admission. The patient eventually died of liver failure at 71 days after admission. Green inclusions are bright green or blue-green inclusions presented in the cytoplasm of neutrophils in Wright-Giemsa stained peripheral blood smears, and is associated with liver failure and high short-term mortality.

Objective To observe the distribution of different types of Helicobacter pylori (H.pylori)infection in non-atrophic gastritis,atrophic gastritis and intestinal metaplasia.Methods From January 2015 to January 2017,457 hospitalized patients with gastritis were collected.All the inpatients received 14C-urea breath test(14C-UBT), H.pylori antibody typing detection, anesthesia gastroendoscopy and pathological analysis of biopsies.According to the results of H.pylori antibody typing,all patients were divided into typeⅠ H.pylori infection group,typeⅡ H.pylori infection group and H.pylori negative group.The relation among different types of H.pylori infection,age and pathological type were analyzed.Analysis of variance and chi-square test were performed for statistical analysis.Results The mean age of patients in typeⅠH.pylori infection group(n=135),typeⅡ H.pylori infection group(n=98)and H.pylori negative group(n=224)in patients with non-atrophic gastritis was(47.5 ± 9.0),(49.0 ± 9.0)and(52.0 ± 11.0)years,respectively, which were lower than those of patients with atrophy gastritis((56.8 ± 10.3),(57.5 ± 12.4)and(62.6 ± 10.4) years,respectively)and patients with intestinal metaplasia((59.2 ± 11.1),(57.5 ± 12.6)and(57.8 ± 10.0)years,respectively),and the differences were statistically significant(F=17.90,6.82 and 18.30,all P<0.01).The proportion of patients with non-atrophic gastritis in H.pylori negative group,typeⅡ H.pylori infection group and typeⅠ H.pylori infection group decreased in turn(56.3%,126/224;40.8%,40/98 and 34.8%,47/135, respectively),and the differences were statistically significant(χ2 =18.44,P<0.01).The proportion of patients with intestinal metaplasia in H.pylori negative group,type Ⅱ H.pylori infection group and type Ⅰ H.pylori infection group increased in turn(23.2%,52/224;26.5%,26/98 and 42.2%,57/135),and the difference was statistically significant(χ2 =11.44,P=0.003).There was no significant difference in the proportion of patients with atrophic gastritis among H.pylori negative group,type Ⅱ H.pylori infection group and type Ⅰ H.pylori infection group(20.5%,46/224;32.7%,32/98 and 23.0%,31/135;χ2 = 5.60,P=0.061).Conclusions Gastric atrophy is easily induced in patients with H.pylori infection.In patients without symptoms,H.pylori should be eradicated before gastric atrophy,which can provide the greatest benefit to the patients.Different types of H.pylori infection have different role in the progress of atrophic gastritis.TypeⅠ H.pylori infection accelerate the progress of atrophic gastritis,especially from atrophic gastritis to intestinal metaplasia,which should be actively eradicated.

Objective To understand the status of infection and epidemic trend of Dengue fever and Chikungunya in Guangdong.Methods Retrospective survey and literature review were used to obtain data on the incidence,etiology of Dengue and Chikungunya.Serological survey was conducted to detect the specific-antibodies in healthy individuals for both Dengue virus (DENV) and Chikungunya virus (CHIKV).Results Three epidemics of Dengue fever were observed during 1990-2012,with the annual incidence rates as 9.75/100 000 in 1995,1.76/100 000 in 2002 and 1.25/100 000 in 2006,respectively.The predominant epidemic strains appeared to be DENV-2 and DENV-4 during 1990-1994.Since 1995,DENV-1 had become the predominant transmission strain which lasted for almost 13 years.Co-existence of multiple serotypes of DENV started in 2009.Of the 7 718 sera from healthy population during 2003-2012,180 specimens were detected positive for specific DENV-IgG antibody,ended with a sero-prevalence rate of 2.33％.All 2 132 sera in 2012 were detected negative for CHIKV-IgG antibody.Conclusion The overall exposure level to Dengue was considered to be low in Guangdong province.However,the predominant transmission mode caused by DENV-1 had been gradually changed into the co-existence of multiple serotypes with the endemic signs appeared in some part of the areas.Chikungunya was a newly emerging disease in Guangdong since local people were lack of basic immunity barrier.Surveillance and control programs thus seemed important.