AIM: In this study, CD147 antibody was used to carry out targeted modification of nanoparticles for protein kinase Cε (PKCε)-siRNA gene therapy to target lung cancer cells.The inhibitory effects of the nanoparticles on the proliferation and invasion of the lung cancer cells were observed.METHODS: The magnetic nanoparticles targeting CD147 protein were assembled as gene vector.The expression of CD147 in the lung cancer cells was observed under laser scanning confocal microscope.The cells were divided into CP group, CN group and LP group as the experimental groups. Targeted nanoparticles were used as CA group.Non-transfected cells were used as control group.The cell transfection was carried out with 250 ng plasmids/well in 6-well plate.The effect of nanocontrast agent on the cell endocytosis was observed under laser scanning confocal microscope.The mRNA expression of PKCε was detected by RT-qPCR.The protein expres-sion of Ki67, MMP3, PKCε, Wnt1 and GAPDH was determined by Western blot.The cell proliferation ability was detec-ted with colony formation assay.The cell invasion ability was detected by Transwell method.RESULTS: The expression of
CD147 protein in the human lung cancer A549 cells was confirmed by immunofluorescence staining.The endocytosis of siRNA into the A549 cells in CP group was observed with the highest efficiency as compared with CN group and LP group. The relative mRNA expression of PKCε in the A549 cells of CP group, CN group, LP group and CA group were (9.76 ± 0.18)%, (98.51 ±0.32)%, (99.17 ±0.16)% and (99.68 ±0.11)%, respectively.The difference between CP group and control group was statistically significant (P <0.05).No significant difference among CN group, LP group and control group was observed.The protein expression of PKCε, Ki-67, MMP3 and Wnt1 in CP group was significantly reduced, and the protein expression levels among CN group, LP group and control group had no significant difference.The colony number in CP group was significantly smaller than that in control group (P <0.05).The effective colony numbers in CN group, LP group and CA group had no significant difference as compared with control group.The number of the invading cells in CP group was significantly less than that in control group (P <0.05).The numbers of the invading cells in CN group, LP group and CA group had no significant difference as compared with control group.CONCLUSION: Nanogene vector targe-ting CD147 can carry PKCε-siRNA to conduct gene therapy efficiently on the lung cancer cells to achieve effective inhibito-ry effects on the proliferation and invasion of the lung cancer cells.

The cloning and identification of frc gene from Oxalobacter formigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacter formigenes was extracted. frc gene fragment was amplified by polymerase chain reaction (PCR) and linked with pEGFP-C1. The recombinant plasmid was designated pEGFP-frc and was identified by restriction-enzyme digestion and sequencing. Human embryo kidney 293 cells were transfected with pEGFP-frc, then RT-PCR and Western blotting were performed to detect the expression of frc gene. The length of frc gene was found to be 1287 bp, and the homology of nucleotides and amino-acid residue with the sequence in GenBank was 95.88% and 99.07%. Bright green fluorescent light could be observed in 293 cells transfected with the pEGFP-frc. frc mRNA and fusion protein FCoAT-EGFP were detected in the cells. It is concluded that frc gene cloned from the Oxalobacter formigenes in the intestines of Chinese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.

Objective To set up the electrochemical determination method for puerarin in Radix Puerariae.Methods The electrochemical behavior and content determination of puerarin were investigated by using coulomb array detector.Results In phosphate buffer with PH 4.32,puerarin showed good linearity in the range of 0.16 ng～ 162 ng.The regression equation was A=1444.7C+ 88.635,r=0.9998(n=7),the average recovery was 96.88 %,and RSD was 0.08 %.Conclusion This method is simple and rapid,and the result is accurate and reliable.It can be used for the determination of puerarin content and biological samples.

Objective This paper aims to find a best position of ephedrine hydrochloride nasal drops by comparing earache occurred in hyperbaric oxygen(HBO )therapy while using ephedrine hydrochloride nasal drops in two positions: laying in lateral position of 45°with head leaning back; laying down flat, so as to supply reference for prevention of earache. Methods We selected 300 cases of HBO patients who were firstly treated by HBO and randomly divided into the observation group and the control group with 150 cases in each group. All these patients were treated by large medical hyperbaric oxygen tank. On the first to third day, ephedrine hydrochloride nasal drops were put into the patients before they were sent into the oxygen tank. Patients in the observation group lay in lateral position of 45°with their head leaning back, and those in the control group were in horizontal position. We put three drops of 1% of ephedrine hydrochloride nasal drops respectively into their nasal cavity. When they came out of the oxygen tank, we asked them if they felt earache and checked their tympanic membrane so as to make a comparison. Results It was found that earache, rate of tympanic membrane injury and the extent of such injury in the observation group was obviously less than that in the control group, there was difference in statistics between these two groups. Conclusions Before HBO treatment, ephedrine hydrochloride nasal drops that put in lateral position of 45°with patients head leaning back can reduce the possibilities of earache occurred in HBO. It was also observed that such position was an ideal one which had anatomical characteristics of Eustachian tube and was safer than laying down flat, so such position is more practical.

The ongoing health reform encourages pharmacists of public hospitals to change their traditional role positioning to satisfy patients’urgent needs in medical dosage guidance.This calls for greater capacity of hospital pharmacy services.Pharmaceutical consultation constitutes a major means of quality service of pharmacists for the people.The paper introduced the standard of establishing the pharmaceutical consultation center and the content of assessment system which are jointly worked out by Beijing Municipal Hospital Authority and the hospitals under the administration of the Beijing Municipal Government,and share the practice & effect in assessing and improving their performance.

Objective To explore the correlation of metabolic syndrome (MS ) with low triiodothyronine(T3)syndrome(LT3S ,or It is known as ESS :euthyroid sick syndrome) ,subclinical hypothyroidism (SCH )and a high serum total homocysteine (tHcy )level (hyperhomocysteinemia )in elderly patients. Methods We retrospectively analyzed thyroid function ,serum glucose ,lipids and homocysteine(Hcy)level in 83 MS patients(MS group) ,30 patients with low T3 syndrome(low T3 group) ,41 patients with SCH (SCH group)and 80 healthy individuals (control group)aged over 60 years from January 2015 to March 2017 in outpatient and inpatient departments of our hospital. The correlations of thyroid function and serum Hcy level with MS components were analyzed by multiple Logistic regression model. Results The incidence of low T3 syndrome and SCH in MS patients was 37.3％ (31/83)and 49.4％ (41/83) ,respectively.Low T3 group versus control group showed the lower levels in free triiodothyronine ,FT4 ,folic acid ,and Hcy ,with statistically significant differences (all P<0.05).SCH group versus control group showed a lower levels of FT 4 ,of Hcy and of folic acid ,but an increased TSH level ,with statistically significant differences (all P<0.05).Patients with TSH >5.5 mU/L had decreased FT3 which occurred later than the rise of TSH (P<0.05) .Univariate correlation analysis showed that FT3 was negatively correlated with Hcy (P< 0.01) ,and TSH was negatively correlated with high-density lipoprotein cholesterol(P<0.01).Multivariate linear regression analysis found linear correlations of TSH with triglyceride ,high-density lipoprotein cholesterol ,and Hcy(all P < 0.05).Logistic regression analysis showed that FT3 ,Hcy and TSH are highly associated with hypertension after adjustment for gender and body mass index (all P< 0.05).Decreased FT3(P<0.01) ,elevated Hcy(P<0.05) ,and increased TSH(P<0.05)were independent risk factors for MS in elderly patients. Conclusions The incidences of low T3 syndrome and SCH are increased with aging. FT3 ,TSH ,and Hcy are independent risk factors for MS in the elderly.

Objective To evaluate the radiosensitization effect of apatinib on esophageal cancer cell line Kyse-150, and to investigate the underlying mechanism. Methods Cells were divided into four groups:control group, apatinib treatment group, X-ray radiation group, and the combination group treated with X-rays plus apatinib. The effect of apatinib with different concentrations on the cell proliferative and radiosensitivity were evaluated by CCK-8 kit and colony formation assay. Flow cytometry method was adopted to detect the effect of apatinib on cell cycle progress and apoptosis induction. Results Apatinib inhibited the proliferation of Kyse-150 cells in time-and dose-dependent manners (r=0. 89-0. 96, P<0. 05). With the increase of apatinib concentration, D0, Dq and SF2 value of Kyse-150 cells decreased and SERD0 value increased. Compared with control group, apatinib alone group, and radiation alone group, the cell apoptosis rate significantly increased in the combination group (t=12. 36, 5. 99, 15. 47,P<0. 05). Compared with control group, the percentages of cells in G2/M phase were all significantly increased in apatinib group, radiation group and combination group (t=8. 81, 39. 69, 20. 61,P<0. 05). Compared with radiation alone group and control group, the percentage of cells in S phase significantly increased in apatinib alone group and combination group(t = 6.06, 3.82,8.81,6.24,P < 0.05). Conclusions Apatinib can increase radiosensitivity of esophageal cancer cell line Kyse-150 possibly by inhibiting cell proliferation, inducing cell apoptosis and causing redistribution of cell cycle.

Objective To evaluate the radiosensitivity effects of apatinib on the esophageal cancer cell line ECA-109 and its cancer stem-like cells,and to investigate the underlying mechanism.Methods A serum-free medium (SFM) was used to culture esophageal cancer stem cell line ECA-109 and enrich the esophageal stem-like spheres.ECA-109 and its stem-like cells were divided into control group,drug treatment group,radiation group and drug plus radiation group.Cell proliferations of ECA-109 and its stem-like cells were detected with CCK-8 method.The concentration of vascular endothelial growth factor (VEGF) in the cell culture medium was determined by enzyme linked immunosorbent assay(ELISA).Cell cycle and apoptosis were detected by flow cytometry method.The expressions of CHK2 and P-STAT3 proteins were detected by Western blot assay.Results With the administration with apatinib for 24,48 and 72 h,the half of the inhibitory concentration (IC50) of ECA-109 stem-like cells was significantly higher than that of the parent cells (t =8.17,9.29,18.85,P ＜ 0.05) in a time dependent manner (parental cells:r2 =0.94-0.97,P ＜0.05;stem-like cells:r2 =0.94-0.98,P ＜0.05).After administration with different concentrations of apatinib (parental cells:10 and 20 μmol/L;stem-like cells:30 and 40 μmol/L) combined with different dose of X-rays (6 and 8 Gy),the proliferations of ECA-109 and its stem-like cells were significantly (t =5.20-39.68,P ＜ 0.05) inhibited compared with radiation alone group.VEGF secretion from both ECA-109 cells and its stem like cells were significantly decreased in different manner (t =7.45,P ＜ 0.05).Compared with control group,the cell apoptosis rate and the percentages of cells in G2/M phase were significantly increased in drug plus radiation group (t =8.83,11.59,P ＜ 0.05),and the expressions of CHK2 and P-STAT3 were decreased in drug group (t =3.36,4.10,P ＜ 0.05).Compared with radiation group,the expressions of CHK2 and P-STAT3 were decreased in drug plus radiation group (t =9.05,2.36,P ＜ 0.05).Conclusions Apatinib enhanced the radiosensitivity of ECA-109 cells and its stem-like cells,which was much more effective on ECA-109 cells and may be related to the radiation-induced inhibition of VEGF signal pathway that can further inhibit cell proliferation,promote cell apoptosis and induce cell cycle redistribution.The higher intrinsic level of VEGF protein may contribute to radioresistance of ECA-109 stem-like cells.

The cloning and identification of frc gene from Oxalobacter formigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacter formigenes was extracted. frc gene fragment was amplified by polymerase chain reaction (PCR) and linked with pEGFP-C1. The recombinant plasmid was designated pEGFP-frc and was identified by restriction-enzyme digestion and sequencing. Human embryo kidney 293 cells were transfected with pEGFP-frc, then RT-PCR and Western blotting were performed to detect the expression offrc gene. The length of frc gene was found to be 1287 bp, and the homology of nucleotides and amino-acid residue with the sequence in GenBank was 95.88% and 99.07%. Bright green fluorescent light could be observed in 293 cells transfected with the pEGFP-frc. frc mRNA and fusion protein FCoAT-EGFP were detected in the cells. It is concluded that frc gene cloned from the Oxalobacterformigenes in the intestines of Chinese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.

Objective:To analyze the relationship between allergen reactivity, atopic disease history and clinical features in patients with chronic inducible urticaria (CIndU) .Methods:A retrospective analysis was conducted on clinical data and follow-up results from 168 patients with CIndU in the Department of Dermatology, Southwest Hospital of Army Medical University from June 2014 to June 2015. Associations were analyzed between allergen reactivity, atopic disease history and clinical characteristics (including patient global assessment [PGA] scores, pruritus intensity, dermatology life quality index [DLQI], proportions of cases with complicated angioedema, natural course, etc.) in patients with different CIndU subtypes. Chi-square test, Mann-Whitney U test and Kaplan-Meier survival analysis were used for statistical analysis. Results:Among the 168 patients with CIndU, 117 were diagnosed with symptomatic dermographism (SD) , 32 with cold contact urticaria (CCU) , 5 with heat contact urticaria (HCU) , and 14 with cholinergic urticaria (CholU) ; there were 46 (39.3%) , 14 (43.8%) , 3, and 9 patients with positive skin prick test (SPT) among the patients with SD, CCU, HCU and CholU respectively, and no significant difference was observed in the positive rate of SPT among patients with different CIndU subtypes ( χ2 = 3.86, P = 0.283) . The SPT-positive CIndU patients showed significantly increased PGA scores, pruritus scores, DLQI scores and proportions of cases with complicated angioedema compared with the SPT-negative patients (all P<0.05) ; the CIndU patients with a personal or family history of atopic diseases also showed significantly increased PGA and DLQI scores compared with those without (both P < 0.05) . For different CIndU subtypes, the pruritus scores, PGA scores, DLQI scores, and proportions of cases with complicated angioedema were significantly higher in the SPT-positive SD patients than in the SPT-negative SD patients (all P < 0.05) ; the DLQI scores were significantly higher in the SPT-positive CholU patients than in the SPT-negative CholU patients ( Z = -2.28, P = 0.019) ; the pruritus scores were significantly higher in the CCU patients with a personal or family history of atopic diseases than in those without ( Z =-2.41, P = 0.022) . Conclusion:The allergen reactivity and atopic disease history of CIndU patients were associated with disease severity, pruritus intensity, quality of life, and the proportion of cases with complicated angioedema, but their relationship with the natural course of CIndU needs to be confirmed by further studies.