1.Clinical application and evaluation of 4 kinds of siphilis serological detection method
Jing YIN ; Guihong XU ; Yan LIU
International Journal of Laboratory Medicine 2014;(23):3244-3245,3248
Objective To compare the value of four kinds of commonly used serological detection method for detecting Trepone-ma Pallidum,i.e.,Treponema Pallidum enzyme-linked immunosorbent assay(TP-ELISA),Treponema pallidum particle agglutina-tion test(TPPA),Treponema Pallidum rapid plasma reagin test(RPR)and Treponema Pallidum antibody detection(TP-AD,emul-sion method).Methods 5 870 specimens from outpatients and inpatients were screened by TP-ELISA.121 cases of detected posi-tive specimen were simultaneously detected by TP-AD,TPPA and RPR.Then the detection results were performed the comparative analysis.Results Among 5 870 specimens,121 cases were positive by ELISA,the detection rate was 2.06%.Among 121 positive cases,119 cases were positive by TPPA,the conformity degree was 98.34%,49 cases were positive by RPR,the conformity degree was 40.41%,113 cases were positive by TP-AD,the conformity degree was 93.38%.With the TPPA results as the standard,there was no statistically significant difference between TPPA and TP-AD(P >0.05),but there was statistically significant difference be-tween TPPA and RPR(P <0.01).Conclusion The four kinds of method have their applicability.ELISA d has good specificity and high sensitivity,and is simple to operate and suitable for the physical examination of large amount of pregnant women,parturients and normal people.TPPA has good specificity and high accuracy,is suitable for definite diagnosis.RPR is suitable for the monito-ring and the curative effect observation in the patients with the active stage of siphilis.Compared with ELISA,TP-AD has slightly less sensitivity,but good specificity and can be used for screening without specific instrument.
2.Effects of total flavonoids of litchi and rosiglitazone on expression of PPAR-γ and CTGF in HSC-T6 rat hepatic stellate cells
Rui LIU ; Guihong CHEN ; Lingli XU ; Weisheng LUO
The Journal of Practical Medicine 2016;32(3):344-347
Objective To investigate effects of total flavonoids of litchi (TFL) on the proliferation of rat hepatic stellate cells (HSC-T6) in comparison with western medicine rosiglitazone, and to explore the mechanism of anti hepatic fibrosis of TFL. Methods Effect of TFL on proliferation of HSC-T6 was examined by MTT. The expression of peroxisome proliferator-activated receptor-γ (PPAR-γ) mRNA, connective tissue growth factor (CTGF) mRNA in HSC-T6 cells exposured were examined by real-time quantitative PCR. Effects on HSC-T6 CTGF protein from TFL and rosiglitazone were detected by Western bloting. Results The expression of PPAR-γ mRNA was upregulated and the expression of CTGF mRNA and protein was downregulated after exposure to TFL and rosiglitazone for 72 hours. And the effect of TFL increased with the increase of concentration. Conclusion TFL can inhibit the proliferation of HSC-T6 and antagonizing liver fibrosis. This mechanism may be associated with the upregulation of PPAR-γ expression and the downregulation of CTGF expression.
3.The expression of the pancreatic duodenal homeobox-1(PDX-1) in pancreas of experimental type 2 diabetic rat and the effect of rosiglitazone on its expression
Caimian XU ; Yuna ZHANG ; Weihua ZHOU ; Guihong LIU ; Tienian ZHU
Chinese Journal of Diabetes 2009;17(11):868-870
Objective To observe PDX-1 expression in the pancreas of type 2 diabetic rat model and effects of rosiglitazone (RGZ) on it. Methods Type 2 diabetic rat model was induced by long-term feeding with high-fat foods followed by a single intraperitoneal injection of low dose of streptozotocin (STZ). The morphological changes of pancreas were examined by microscopy and immunohistochemistry. The mRNA levels of PDX-1 and insulin were determined by RT-PCR. PDX-1 protein expression was detected with Western blot. Results The percentage of insulin-positive cells(12.75±2.18),the average optical densities of PDX-1-positive area(0.240±0.051),PDX-1 mRNA level (0.153±0.071)and PDX-1 protein level(0.253±0.028) were significantly lower in untreated diabetic rats than in the controls(42.61±2.68,0.648±0.087,0.49±0.032,0.720±0.036 respectively)(all P<0.01),and in RGZ-treated versus untreated group those parameters increased significantly (all P<0.05~0.01). Conclusions Rosiglitazone could protect islet cell function,and improve PDX-1 expressions in both mRNA and protein levels.
4.Reconstruction of anti-hepatocellular carcinoma scFv pharmacy and its killing effect on hepatocardnoma cell in vitro
Guihong ZHANG ; Yanfang LIU ; Yong FU ; Haiyang HU ; Dawei CHEN
Cancer Research and Clinic 2008;20(12):802-804
objective To observe whether the killing effect on HCC SMMC-7721 cell of the antihepatocellular carcinoma scFv reconstructed by pharmacy was enhanced or not.Methods Prokarycytic expression vector containing PET32a-RC-RNase was induced to express by IPTG.The inclusion body purified and Western-blotting was used.PC.CHOL and CHS was added in chloroform.Dry membrane was formed after chloroform was removed.RC-RNase protein solution was added to dissolute the membrane.Then pass the solution over a Sephadex G-50 column after ultrasound and filtrated to detect the encapsulation efficiency of the liposome.The solution reacted in EDC.SSNHS and MES for 30 minutes.Then add hdscFv to the solution in 4 ℃ over night.MTT method was used to detect the killing effect on HCC cell of immunoliposome RC-RNase,immunotoxin RC-RNase and liposome RC-RNase in vitro.Resuits The killing effect on HCC cell of immunoliposome RC-RNase is the best.but that of Iiposome RC-RNase is the worst.The respective JC50 are:3.28μg/ml,22.44μg/ml and 98.26μg/ml.Conclusion The anti-hepatocellular carcinoma scFv relomtructed by pharmacy can promote the killing effect on HCC cell and may have potential in the treatment of hepatocarcinoma.
5.Study on the cognition status of implementing case-based payment policy among healthcare providers:A case in Chengdu, Sichuan Province
Guihong TAN ; Xiaohui REN ; Zhijun LIU ; Ningxiu LI ; Jing WANG
Chinese Journal of Health Policy 2014;(4):38-43
Objective:To understand the implementation status of case-based payment among healthcare provid-ers and their cognition on the matter. Methods:Semi-structured interview was conducted on 30 purposely selected staff from 9 hospitals in Chengdu. Results:After one-year implementation of case-based payment, hospitals at different level carried out the policy vigorously:executed corresponding expense control measures and management. Neverthe-less, the proportion of cases that were paid with case-based payment was low, moreover the inclusion criteria for case was of disunity and the formulation of the expense standard was ambiguous to some extent. Conclusion:Certain achievements were accompanied with problems, so it is essential to refine reimbursement standards, improve case-based payment, make clear the inclusion criteria for case and extend the covering range of case-based payment in Chengdu. In addition, the medical insurance agency should strengthen the supervision of healthcare providers,and guide them to set up effective incentive mechanism.
6.Immunotherapy experimental study of γδT cells from human peripheral blood on human liver cell nude mouse xenograft in vivo
Li XIA ; Yanfei CHI ; Xiaofeng LI ; Juan WANG ; Bailin LIU ; Guihong HAN ; Yanhang LIU ; Yuxia LIU
Chinese Journal of Immunology 2017;33(5):702-706
Objective:To study the immunotherapy effects of different doses of human peripheral blood γδT cells on human hepatoma cells (SMMC-7721) xenograft model.Methods: (1)The nude mouse model of liver cancer was established by inoculated BALB/c mouse subcutaneous with human hepatoma cell line (SMMC-7721).(2)The mononuclear cells in healthy human were extracted from peripheral blood,and specific amplification γδT cells in vitro.(3) The nude mouse model divided into 5 groups by random.The positive control group was 5-Fu,negative control group was normal saline(NS).The treatment group was injected different doses of γδT cells(1×105,5×105 and 25×105)by nude mice tail vein.The positive control group injected 5-Fu by enterocoelia,negative control group injected NS by tail veins.The inhibition effect of different dose γδT cells on tumor was observed,including weight,food intake and growth conditions,etc.and the changes of tumor volume (TV),relative tumor volume (RTV)and relative tumor appreciation rate[T/C(%)] were compared with positive control group and negative control group.Results: Different dose of γδT cells had different degree of inhibition on nude mouse xenograft growth.RTV compared with saline negative control group was statistically significant (P<0.05).Compared with the positive control group of 5-Fu,the TV growth was significantly lower than the 5-Fu,degree of inhibition was similar in RTV each dose group,and all slightly higher than the 5-Fu positive control group.The each dose group of T/C (%)was slightly lower than the relative tumor proliferation rate of the control group of 5-Fu,but had no significant difference.Conclusion: The γδT cells from peripheral blood had significant inhibitory effect on nude mice transplanted liver tumor and it may be used as a new treatment for liver cancer immunotherapy provide experimental data.
7.Improvement of the cholinergic function by melatonin in amnesic rats induced by amyloid ?-peptide 25~35
Yuxian SHEN ; Wei WEI ; Guihong ZHANG ; Chao LIU ; Lihua LIU ; Shuyun XU ;
Chinese Pharmacological Bulletin 1987;0(03):-
AIM To investigate whether melatonin improve the learning and memory dysfunction in the amnesic rats induced by amyloid ? peptide 25~35 (A? 25~35 ) via cholinergic nervous system or not.Methods The amnesic model in adult rats was induced by injection of A? 25~35 into hippocampus; Morris water maze was used to determine the effects of A? 25~35 and melatonin on the learning and memory. The activity of the choline acetyltransferase and acetylcholinesterase were determined by immunohistochemistry and spectrophotometry respectively. RESULTS Injection of A? 25~35 20 ?ginto the adult rats hippocampus induced learning and memory dysfunction, and a decrease in the number of ChAT immunoreactive neurons in hippocampus. Melatonin (0 1, 1, and 10 mg?kg -1 , ig?10 d) improved the A? 25~35 treated rats cognitive function and increased the number of ChAT immunoreactive neurons in hippocampus. CONCLUSION Improvement of the cholinergic dysfunction by melatonin in adult rats induced by amyloid ? peptide 25~35 may be via cholinergic nervous system.
8.Age features of pure leukoaraiosis in non-demented outpatients
Xueying YU ; Guihong WANG ; Xiaoting GUAN ; Yun LIU ; Suxiang WANG ; Zhizhong LIU ; Yongjun WANG
Chinese Journal of Internal Medicine 2010;49(6):488-490
Objective To investigate the aging features of pure leukoaraiosis (LA) in nondemented outpatients. Methods The outpatients with age older than 40 years, without taking cholesterol lowering and B vitamin medications and with mini-mental state examination more than 24 scores were selected from July 2008 to December. 2009 in Beijing Tiantan Hospital. LA was defined with MRI. Patients were classified into two groups i. e. LA group consisting of 138 patients with leukoaraiosis but without lacunar lesions and cortical infarcts and a control group consisting of 124 patients without any lesion in brain. Age and other vascular risk factors were also investigated. Results Age of the patients in the LA group was significantly higher than that in the control group (P< 0.001). Multivariable logistic regression analysis showed that age was independently associated with pure LA ( OR 1.080, 95% CI 1.042-1.120), after adjusting sex, vascular risk factors and presence of atherosclerosis in cervical arteries. If age-stratification was further considered, logistic regression analysis showed that OR (95% CI) for LA was 2.693 (95% CI 1.103-6.575) in a 60-69 year group and 13. 527(95%CI 3.319-55.131) in a≥70 year group as compared with a 40-49 year group. Conclusion Age is a determining risk factor for pure LA and patients with age older than 60 years are at high risk of LA.
9.An experimental study on dexamethasone combined with puerarin in treatment of acute paraquat-poisoned rats
Guihong CHEN ; Shiping LIU ; Xiaoping CAO ; Wu ZHONG ; Muhu CHEN ; Wen ZHANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2016;23(5):499-503
ObjectiveTo explore the therapeutic effect and mechanism of dexamethasone combined with puerarin for treatment of paraquat (PQ) poisoned rats.Methods Thirty Sprague Dawley (SD) rats were divided by random number table into 5 groups: control, model, dexamethasone, puerarin and combined groups, 6 rats in each group. The PQ poisoned rat model was established by intraperitoneal injection of PQ 25 mg/kg (1 mL), while in the rat of the control group, the same volume of normal saline was injected intraperitoneally. After 2 hours, in the rat of dexamethasone group, 20 mg/kg dexamethasone in 1.5 mL was injected intraperitoneally, and in the rat of puerarin group, 100 mg/kg puerarin in 1.5 mL was injected intraperitoneally. In the rat of combined group, 20 mg/kg dexamethasone combined with 100 mg/kg puerarin in a total amount of 1.5 mL was injected intraperitoneally. In the control group and model group, 1.5 mL of normal saline was injected intraperitoneally. The above mentioned treatments were repeated once more 26 hours later. In 26-50 hours after modeling, urine was collected, and urine N-acetyl-beta-D-amino glycosidase enzymes (NAG) was tested; at the 50th, abdominal aortic blood was collected to test oxygen partial pressure (PaO2). The lung index, kidney index, and the levels of malonaldehyde (MDA), superoxide dismutase (SOD) and myeloperoxidase (MPO) were measured in the left lung and kidney tissue homogenates separately; the right lung and kidney were
harvested to observe pathological changes under light microscope. Another 30 SD rats were treated the same as above measures but no sacrifice to observe 30-day survival rate in each group.Results The lung index, kidney index, NAG in urine, MDA and MPO levels in lung and kidney tissue homogenates in model group were significantly higher than those in the control group [lung index: 9.80±1.83 vs. 4.97±1.14, kidney index: 9.40±1.32 vs. 7.01±0.32, NAG (U·kg-1·day-1): 1.93±0.18 vs. 0.41±0.03, MDA of lung (nmol/mg): 1.04±0.15 vs. 0.28±0.10, MDA of kidney (nmol/mg): 1.39±0.16 vs. 0.66±0.13, MPO of lung (U/g): 1.14±0.08 vs. 0.81±0.06, MPO of kidney (U/g): 0.88±0.08 vs. 0.52±0.12]; while PaO2 [mmHg (1 mmHg = 0.133 kPa): 59.83±4.40 vs. 97.00±2.83] and SOD (U/mg): lung was 27.38±3.48 vs. 86.88±5.88; kidney was 24.18±3.74 vs. 57.86±6.14) were obviously lower than those in control group (allP < 0.05). After drug treatment was given, lung index (7.21±1.87), urine NAG (1.01±0.21) and MDA (lung was 0.49±0.09, kidney was 0.85±0.08), MPO (lung was 0.97±0.07, kidney was 0.68±0.10) in the puerarin group were significantly lower than those in model group, while PaO2 (82.17±5.38), SOD (lung was 68.99±6.51, kidney was 37.90±3.62) were remarkably higher than those in model group (allP < 0.05). The improvement in the indexes related to the kidney injury was not obvious in dexamethasone group and in the dexamethasone combined with puerarin group, while the lung and kidney pathological changes were lesser in extent in each of the above two treatment groups than those in model group. The 30-day survival rate in model group was significantly lower than that in control group (0% vs. 100.0%);the 30-day survival rates in control group, dexamethasone group, puerarin group and combined group were remarkably higher than those in model group (100.0%, 16.7%, 50.0% and 33.3% vs. 0%, allP < 0.05). But there were no statistical significant differences in the survival rates among those treatment groups (P > 0.05).Conclusions Dexamethasone can improve the prognosis of rats with acute paraquat intoxication, it can provide lung protection markedly, but cannot provide significant protective effect on kidney; puerarin has therapeutic effect on rats with acute PQ poisoning, it can provide not only lung protection but also kidney protection. The effect of treatment with dexamethasone combined with puerarin is not superior to that by using dexamethasone or puerarin alone.
10.The inhibitory effect of isoquercitrin onRaf/MEK/ERK signaling pathway in HepG2 cells
Guojun JIANG ; Tianxu LIU ; Guihong HUANG ; Ting WU ; Liqun TAO ; Zhaoming ZHU
Chinese Pharmacological Bulletin 2017;33(10):1382-1387
Aim To study the inhibitory effect of isoquercitrin on Raf/MEK/ERK signaling pathway in HepG2 cells.Methods MTT was used to detect the proliferation of human liver cancer HepG2 cells after the treatment of isoquercitrin.The morphology and growth of cells were observed under inverted microscope after the different concentrations of isoquercitrin(0, 40, 80, 160, 320 μmol·L-1) to treat HepG2 cells for 24 and 48 h.Cell cycle was assessed by flow cytometry.Ras, Raf, MEK, ERK expression was assayed by Western blot, and mRNA expression was detected by quantitative fluorescence PCR.Results Isoquercitrin could inhibit the growth of HepG2 cells in a concentration-and time-dependent manner.Typical morphological changes of apoptosis were observed by inverted microscopy after HepG2 cells were treated with different concentrations of of isoquercitrin for 24 h or 48 h.The cell cycle assay showed that with the increasing concentration of isoquerditrin, the number of cells that was arrested in G1 phase gradually increased.Compared with the blank group, the expressions of Ras, Raf, MEK, ERK mRNA were down-regulated, and related proteins expression were also down-regulated(P<0.05), and these results had statistical significance.Conclusion Isoquercitrin can induce the apoptosis of HepG2 cells, which may be related to the Raf/MEK/ERK signaling pathway.