AIM: To explore the feasibility of using vector-based small interfereing RNA(siRNA) to inhibit the expression of MDR1 mRNA and P-glycoprotein and to reverse the multidrug resistance of drug-resistant ovarian cancer cell line.METHODS: An adriamycin-resistant human ovarian cancer cell subline OVCAR/AR was established by stepwise inducement.Another mutidrug-resistant human ovarian cancer cell subline OVCAR/MDR was established by transfecting multidrug resistant gene 1(MDR1) into ovarian carcinoma cell line OVCAR-3.Transfection of MDR1 mRNA specific siRNA expressing plasmids(pSN/mdr1a and pSN/mdr1b) into OVCAR/AR and OVCAR/MDR cells was performed using liposome transfection reagents.MDR1 mRNA expression level was quantified using real time reverse transcription polymerase chain reaction(real time RT-PCR).Flow cytometry(FCM) was performed to assess the expression of p-glycoprotein(P-gp).Multidrug resistant to anticancer agents was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide(MTT) assay.RESULTS: Basal MDR1 mRNA expression level in drug-resistant cell line OVCAR/MDR was higher than that in OVCAR/AR cell line,and was both higher than that in its parent cell line OVCAR-3.The expression of MDR1 mRNA and P-gp protein in both OVCAR/AR and OVCAR/MDR cells was inhibited dramatically by transfecting with pSN/mdr1a and pSN/mdr1b.The reversal rate of chemoresistance to adriamycin in OVCAR/AR and OVCAR/MDR transfected with pSN/mdr1a and pSN/mdr1b was 79.5% and 93.9%,compared with control group.CONCLUSION: MDR1 expression in the drug-resistant cell lines is partially inhibited by treatment with vector-based MDR1 specific small interference RNAs at the mRNA and protein level,which increases the chemotherapy sensitivity of these drug resistant ovarian carcinoma cell sublines.

Objective:To investigate the influence of fetal liver AFT024 cells on the transfection efficiency of multidrug resistant gene 1(MDR1)and the in vitro expansion of CD34+ cells derived from umbilical cord blood.Methods:CD34+ cells were isolated from human umbilical cord blood by MACS CD34 Progenitor Cell Isolation Kit and co-cultured with AFT024 cells(AFT024 group)or cultured alone(control group)for 7 days.During the subsequent 14 days,retrovirus carrying MDR1 gene was supplemented twice a week to transfect CD34+ cells.On the 7th,14th and 21st day after culture,the number of total nucleated cells(TNC)was counted,the ratio of CD34+ cells was assayed by flow cytometry(FCM)and the number of CD34+ cells was calculated,and colony-forming cells(CFC)were counted by methylcellulose cultures.RT-PCR method was used to detect the level of MDR1 mRNA in the transfected cells.The expression and function of P-glycoprotein(P-gp)were evaluated by FCM assay and Rhodamine-123 efflux assay,respectively.The gene transfection efficiency was calculated by drug-resistant colony-forming cells assay.Results:(1)The MDR1 mRNA level in AFT024 group than that in control group.The gene transfection efficiency in AFT024 group was significantly higher than that in control group(46.0% vs 15.2%,P0.05).On the 14th day,the expansion fold of TNCs in control group was significantly higher than that in AFT024 group(P0.05).The expansion folds of CD34+ cells and CFCs in the AFT024 group were significantly higher than that of the control group(P

Aliquat 336 functionalized chelating adsorbent derived from chitosan for enrichment and separation of Y(Ⅲ) were investigated by static adsorption method. The adsorption of Y(Ⅲ) was greatly influenced by the pH of solution, and reached maximum at 20 ℃ using 90 mg/L Y(Ⅲ) at pH 4. 9, and the adsorption of Y(Ⅲ) followed a pseudo-second-order kinetics model and the Langmuir isotherm model. The reduction of Y(Ⅲ) adsorption with the increasing of temperature meant that the adsorption process was exothermic. XPS analysis demonstrated that both cations and anions of the adsorbent were involved in adsorption process, thereby resulting in an improved adsorption of Y(Ⅲ). The adsorbent was thus efficient for enrichment and separation of rare earths from waste rare earth phosphor.

OBJECTIVE:To systematically review the efficacy and safety of cinacalcet in the treatment of hemodialysis pa-tients with secondary hyperparathyroidism,and provide evidence-based reference for the clinical treatment. METHODS:Retrieved from Medline,Cochrane Library,EMBase and CBM,randomized controlled trials(RCT)about cinacalcet in the treatment of he-modialysis patients with secondary hyperparathyroidism (SHPT) were collected. Meta-analysis was performed by using Rev Man 5.3.5 software after data extract and quality evaluation by Cochrane systematic Rev Man 5.3.5. RESULTS:Totally 7 RCTs were en-rolled,involving 1 987 patients. Results of Meta-analysis showed cinacalcet can significantly reduce the rate of surgical parathyroid-ectomy[RR=0.23,95%CI(0.06,0.89),P=0.03],incidence of fracture[RR=0.26,95%CI(0.12,0.60),P=0.002] and increase the incidences of hypocalcemia[RR=9.81,95%CI(3.92,4.59),P<0.001],nausea[RR=1.97,95%CI(1.58,2.46),P<0.001] and vomit-ing[RR=1.91,95%CI(1.50,2.42),P<0.001],while it showed no significant effect on the the incidence of all-cause mortality and cardiovascular death. CONCLUSIONS:The clinical efficacy of cinacalcet in the treatment of hemodialysis patients with secondary hyperparathyroidism is good,but there are common adverse reactions such as nausea and vomiting,hypocalcemia.

Objective: To investigate the changes and clinical significance of vascular endothelial (VE)-cadherin and procalcitonin (PCT) in serum and cerebrospinal fluid (CSF) of children with viral encephalitis or bacterial meningitis(BM). Methods: A total of 42 cases of children with viral encephalitis(viral encephalitis group), 36 cases of children with BM(BM group), and 20 cases of children with non-nervous system injury(control group) were selected from September 2016 to June 2018 at the Third Hospital of Zhengzhou University.The serum and CSF levels of VE-cadherin and PCT levels of the 3 groups were detected by using enzyme-linked immunosorbent assay. Results: The levels of VE-cadherin in the serum of viral encephalitis group, BM group and control group at the acute phase were (5.60±1.17) mg/L, (7.08±1.01) mg/L and (2.52±0.68) mg/L respectively, and the levels of VE-cadherin in CSF of viral encephalitis group, BM group and control group were (6.00±1.09) mg/L, (6.97±1.11) mg/L and(1.93±0.88) mg/L, respectively.The levels of PCT in the serum of viral encephalitis group, BM group and control group at the acute phase were (0.26±0.11) μg/L, (0.82±0.17) μg/L and (0.27±0.13) μg/L, respectively, and the levels of PCT in the CSF of viral encephalitis group, BM group and control group were (0.25±0.11) μg/L, (0.72±0.14) μg/L, (0.28±0.17) μg/L, respectively.As a result, the levels of VE-cadherin and PCT in the serum and CSF of BM group showed significant increase, compared with viral encephalitis group and control group in the acute phase(F=124.94, 163.21, 151.62, 127.37, all P<0.01). The levels of VE-cadherin in the serum and CSF of viral encephalitis group were also significantly higher than that of control group (all P<0.01), but there was no difference between viral encephalitis group and control group about the levels of PCT in the serum and CSF (all P>0.05). The levels of VE-cadherin in the serum of viral encephalitis group and BM group after treatment were (2.34±0.81) mg/L and (2.67±1.29) mg/L, and were(2.55±0.92) mg/L and(2.39±0.74) mg/L in the CSF.The levels of PCT in the serum of viral encephalitis group and BM group after treatment were (0.25±0.11) μg/L, (0.30±0.17) μg/L, and the levels of PCT in the CSF of viral encephalitis group and BM group were(0.22±0.10) μg/L and (0.27±0.12) μg/L.After effective treatment, the levels of VE-cadherin, PCT in serum and CSF of BM group and viral encephalitis group were almost equal, and the difference was not statistically significant(F=1.83, 0.76, 2.72, 3.89, all P>0.05). In the receiver operating characteristic (ROC) curve, the area under the ROC curve of VE-cadherin in serum and CSF was 0.896 and 0.912, and was 0.670 and 0.668 of PCT respectively. Conclusions VE-cadherin may be involved in the early stage of intracranial infection, and it may be helpful in differentiation of virus or bacterial infection with PCT.VE-cadherin has a good diagnostic value for intracranial infection.

OBJECTIVETo investigate the impact of long-time and short-time blood pressure variability (BPV) on glomerular filtration rate (eGFR) in elderly population.

METHODSA total of 2 464 participants aged of ≥60 years old without history of stroke and coronary heart disease were selected with random sampling method from the individuals underwent health check up in Tangshan Kailuan Hospital, Kailuan Linxi Hospital, Kailuan Zhaogezhuang Hospital between 2006 and 2013. The study participants were asked to join a face-to-face interview every two years. Long-time BPV was defined as the standard deviation of all SBP values at the baseline visit and following visits, short-time BPV was defined as the standard deviation of day time blood pressure and night time blood pressure which was derived from 24 hours ambulatory blood pressure monitoring. Multivariate linear regression models were used to test the impact of long- and short-time BPV on eGFR.

RESULTSThe study included 3 participants groups including the long-time SBPV group (2 279 participants), the short-time SBPV group (1 636 participants) and the long- plus short-time SBPV group (1 632 participants). Participants were further sub grouped by median value (NO.1median value). eGFR in the long-time SBPV NO.1 and NO.2 group was 83.19 and 81.49 ml·min(-1)·1.73 m(-2) respectively, in the short time SBPV NO.1 and NO.2 group was 83.53 and 80.81ml·min(-1)·1.73 m(-2) of the day time, and was 83.20 and 81.14 ml·min(-1)·1.73 m(-2) of the night time, respectively. eGFR in the long- plus short-time SBPV NO.1 and NO.2 group was 83.21 and 81.08 ml·min(-1)·1.73 m(-2) of the long-time SBPV, and was 83.53 and 80.75 ml·min(-1)·1.73 m(-2) of the day time, and was 83.18 and 81.11 ml·min(-1)·1.73 m(-2) of the night time. Significant linear relationship was found between higher day time SBPV and lower eGFR (P<0.05). After adjusting for confounding parameters including age, sex, body mass index, 1 mmHg(1 mmHg=0.133 kPa) of day time SBPV increase was related 0.21 ml·min(-1)·1.73 m(-2) eGFR decrease (P<0.05).

CONCLUSIONThe higher day time SBPV is associated with reduced eGFR in the elderly population.Clinical Trail Registry Chinese Clinical Trial Registry, ChiCTR-TNC-11001489.

Aged ; Blood Pressure ; Blood Pressure Monitoring, Ambulatory ; Coronary Disease ; Glomerular Filtration Rate ; Humans ; Time Factors

Objective The monitoring of the feeding situation of active pharmaceutical ingredient in Shendan-Chenpi tablet by HPLC-MS/MS,and the content of sodium taurocholate and hesperidin.Methods Using ZORBAX Eclipse Plus C18(4.6 mm×250 mm, 5 μm)Column temperature 40 ℃, Mobile phase with acetonitrile-10 mmol/sodium acetate solution, using gradient elution program. Active ingredients were separated by HPLC, and the Electrospray Ionization Mass (ESI) source was applied and operated in the negative ion mode, and reactions ion monitoring mode(MRM) for quantitative analysis were selected. Results The proprietary Chinese medicine is judged by the prescription feeding process, through analysis and contrast of the medicinal materials, reference substance of primary mass spectrogram, secondary mass spectrogram of peak. The aurocholic acid sodium and hesperidin had a good linear relationship in 0.242×10-2-1.45×10-2μg(r=0.996 0), 0.688×10-2-10.30×10-2μg(r=0.999 2), and the precision test were 0.78% and 1.56%, and the recovery rate were 102%-103%,96%-109%. Conclusions The method was simple, accurate and reliable, high sensitive and fast. The comprehensive monitoring was applied to the Shedan-Chenpi tablet in feeding analysis and quality.

Objective The HPLC-MS/MS is used to comprehensively monitor the feeding conditions of raw materials in Yuanhu analgesic capsules, and the content of the index components can be detected at the same time. Methods The Inertsil ODS-3 (4.6 mm×250 mm, 5 μm) was used with the column temperature 40 ℃, Flow phase: 0.1% formic acid-acetonitrile; the gradient elution program, active ingredients were separated by HPLC, and the Electrospray Ionization Mass (ESI) source was applied and operated in the negative ion mode, and reactions ion monitoring mode (MRM) for quantitative analysis were selected. Results Through analysis and contrast of the medicinal materials, reference substance of primary mass spectrogram showed the same characteristic peak, and the proprietary Chinese medicine can be judged by prescription feeding process. The tetrahydroxene and imperatorin had a good linear relationship in 5.08×10-5-30.45×10-5μg (r=0.999 4), 5.02×10-5-30.09×10-5μg (r=0.999 2). Precision test were 0.99% and 1.14%, the recovery rate were 97.02%-99.66%, 97.62%-99.94%. Conclusions The method is simple, accurate and reliable, high sensitive and fast. It is suitable to monitor the feeding condition and quality of yuanhu analgesic capsules.

Objective: To investigate the changes and clinical significance of Caveolin-1, matrix metalloproteinase-9(MMP-9) and interleukin-1β(IL-1β)in cerebrospinal fluid of children with bacterial meningitis or viral encephalitis. Methods: Thirty-six cases of children with bacterial meningitis, 42 cases of children with viral encephalitis, and 20 cases of children with non-nervous system infection were selected from September 2016 to June 2018 at the Third Affiliated Hospital of Zhengzhou University.The levels of Caveolin-1, MMP-9 and IL-1β in cerebrospinal fluid were detected by using enzyme linked immunosorbent assay(ELISA). Results: Cerebrospinal fluid Caveolin-1, MMP-9 , IL-1β levels in the acute phase of bacterial meningitis were(49.06±8.96) ng/L, (134.79±18.88) μg/L, (100.02±14.67) μg/L, respectively, and (29.13±7.25) ng/L, (18.69±7.23) μg/L, (47.57±8.95) μg/L in recovery phase, which were higher than those of the controls[(11.18±2.24) ng/L, (11.53±3.54) μg/L, (39.75±7.08) μg/L)], and the differences were significant (all P<0.05). Cerebrospinal fluid Caveolin-1, MMP-9, IL-1β levels in the acute phase of viral encephalitis were (42.71±10.48) ng/L, (62.78±17.39) μg/L, (57.97±11.28) μg/L, respectively, and (29.13±7.25) ng/L, (18.69±7.23) μg/L, (47.57±8.95) μg/L in recovery phase, which were higher than those of controls, and the differences were significant (all P<0.05). The levels of Caveolin-1, MMP-9 and IL-1β in cerebrospinal fluid of bacterial meningitis group and viral encephalitis group were significantly higher than those of convalescent group (all P<0.05). The levels of Caveolin-1, MMP-9, IL-1β in cerebrospinal fluid of bacterial meningitis group were significantly higher than those in viral encephalitis group (all P<0.05) in the acute phase, and no significant difference was found in the recovery phase(all P>0.05). Cerebrospinal fluid Caveolin-1, MMP-9, IL-1β showed no significant difference among children with different severity of intracranial infection.Correlation analysis showed that there was a positive correlation between Caveolin-1, MMP-9 and IL-1 β levels in cerebrospinal fluid of acute in bacterial meningitis group and viral encephalitis group(Caveolin-1 and MMP-9: R²=0.239, P<0.05; MMP-9 and IL-1β: R²=0.766, P<0.01; Caveolin-1 and IL-1β: R²=0.245, P<0.05). Conclusions Caveolin-1, MMP-9 and IL-1 β involved in the pathogenesis of intracranial infection in children, and the effects of different pathogens on intracranial infection were different.

BACKGROUNDLymphocyte function and homeostasis is associated with immune defence to infection. Apoptosis of lymphocytes might be a considerably important component which has an impact on immunity to infections in people with hyperglycemia. The aim of this study was to explore the mitochondrial apoptosis pathway of lymphocyte in diabetic patients.

METHODSSixty patients with type 2 diabetes mellitus and fifty healthy volunteers were included in this study. Annexin V and propidiumiodide (PI) were joined in the isolated lymphocytes and the rate of lymphocyte apoptosis was calculated with flow cytometry. Observation of the lymphocytes was done using transmission electron microscopy; mitochondria had been extracted and then mitochondrial membrane potential (MMP) was detected to assess mitochondrial function; the mRNA level of Bcl-2, cytochrome c (Cyt-C), caspase-9 and caspase-3 were analyzed by real-time reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSApoptosis rate of lymphocyte was significantly higher in diabetic group than that in normal control group (P < 0.05). Transmission electron microscopy showed lymphocyte shrinkage and breakage, chromatin condensation and less mitochondria; a fall in MMP levels was also evident; Bcl-2 concentration was reduced and the expressions of caspase-9, caspase-3 and Cyt-C were elevated (P < 0.05) in diabetic patients.

CONCLUSIONSThe rate of lymphocyte apoptosis was significantly higher in type 2 diabetic patients than that in normal population. Mitochondrial apoptosis pathway may play a very important role in decreasing function of lymphocyte in diabetes.

Aged ; Aged, 80 and over ; Apoptosis ; physiology ; Caspase 3 ; genetics ; Caspase 9 ; genetics ; Diabetes Mellitus, Type 2 ; metabolism ; Female ; Humans ; Lymphocytes ; cytology ; metabolism ; ultrastructure ; Male ; Microscopy, Electron, Transmission ; Mitochondria ; metabolism ; Real-Time Polymerase Chain Reaction ; bcl-2-Associated X Protein ; genetics