1.Development of a time-resolved fluorescence immunoassay for rapid determinat-ion of levels of cardiac troponin Ⅰantigen in human serum
Jie ZHANG ; Lingling LU ; Xiaofu PAN ; Lilin ZOU ; Jimin GAO
Chinese Journal of Immunology 2014;(8):1093-1097
To establish a method to detect cardiac troponin I by using time-resolved fluoroimmunoassay ( TRFIA) and apply to the clinic.Methods:The assay were measured by TRIFA and double antibody sandwich method .Standard protocols were evaluated with the standard curve , the limit of detection , stability, precision and cross reaction .Healthy reference populations and clinical serum specimens were measured to established the reference interval and evaluated the perspective of the clinical application . Results:The standard curve was Y=7485 .878+1400.924 X with a correlation coefficient of 0.999.The limit of detection was 0.052 ng/ml.The intra-and inter-assay coefficients of variation ( CV) were all <10%.Reference values was <0.14 μg/L.The AUC of ROC curve was 0.971 while the sensitivity was 96.45%,the specificity was 91.43% and the accuracy was 95.69%, with 98.45% of positive predictive value and 82.05%of negative predictive value.The correlation coefficient was 0.993 between our proposed method and the commercially available CLIA kits.There was no significant difference in statistics compared with ECG , CK-MB and cTnT ( P>0.05 ).There was significant difference in statistics compared before and after treatment with AMI ( P<0.001 ) .Conclusion: The TRFIA method for detecting cTnI achieves clinical application standards and may be used for the diagnosis and serosurveillance of acute myocardial infarction patients.
2.Correlation of Active Ingredients in the Roots of Paridis rhizoma Genus Plants with Mycorrhizal Infection Rate and Soil Nutrients
Xingjiao PAN ; Jie ZHANG ; Fengzhong LU ; Bo DING ; Nong ZHOU
China Pharmacy 2016;27(22):3037-3040
OBJECTIVE:To investigate the correlation of total saponins and total polysaccharides content in the roots of Pari-dis rhizoma genus plants with Arbuscular mycorrhizal fungi(AMF)infection rate and rhizosphere soil nutrients. METHODS:Tak-ing 16 pieces of Paridis rhizoma herbs and rhizosphere soil in Three Gorges Reservoir Area as the research object,mycorrhizal in-fection rate and infection intensity,the contents of total saponins and total polysaccharides in roots and stem,and rhizosphere soil nutrient contents were all determined. SPSS18.0 software was adopted for data multiple comparison and correlation analysis. RE-SULTS:Paridis rhizoma genus plants roots in Three Gorges Reservoir Area could form a good symbiotic relationship with AMF. In-fection rate ranged from 85.19% to 99.80%,and infection intensity ranged from 16.55% to 72.27%. Paridis rhizoma from differ-ent origins and varieties had significant difference in the contents of total saponins and total polysaccharides (P<0.05). Soil pH was moderate,the average content of total nitrogen was in insufficient level,the average concentration of the remaining nutrients belonged to middle levels or above. Correlation analysis showed that the mycorrhizal infection rate and infection intensity of Paridis rhizoma were not significantly correlated with its quality. The total saponins were significantly negatively correlated with organic matter and available nitrogen. The total saponins and total polysaccharides were positively correlated with available phosphorus,po-tassium and pH value,but they were not significantly correlated with the other soil nutrients. CONCLUSIONS:Paridis rhizoma is selective to different soil nutrients. Rich soil available phosphorus and potassium supply are in favor of the synthesis and accumula-tion of total saponins and total polysaccharides in Paridis rhizoma roots.
3.Antitumor Activity of Dichloromethane Extract from Salvia plebeia and Induction of Apoptosis on K562 Cells
Jie REN ; Shasha PAN ; Xuzhang LU ; Min ZHOU ; Kun HU
Chinese Herbal Medicines 2011;03(1):36-40
Objective To study the antitumor activity of extract from Salvia plebeia and investigate whether the extract induce apoptosis of K562 cells. Methods The aqueous, petroleum ether, dichloromethane (CH2Cl2), ethyl acetate, and butanol extracts were prepared from the aerial parts of 5. plebeia. Taking fluorouracil as reference, the cytotoxic activities of these extracts on HeLa, A549, SGC-7901, HCT-116, K562, LoVo, DU-145, and HepG2 cells were evaluated. To clarify the apoptosis of K562 cells induced by CH2Cl2 extract, the methods of Hoechst 33258 staining, flow cytometry assay, and DNA ladder assay were investigated. Results The CH2Cl2 extract showed the most potent cytotoxic effect against K562 cells, with an IC50 < 15 μg/mL for 3 d treatment. The characteristic apoptotic symptoms such as DNA fragmentation and chromatin condensation were also observed in the K562 cells. Conclusion The CH2Cl2 extract from S. plebeia may inhibit the cancer cell proliferation by inducing cell apoptosis.
4.The adverse reaction of dexmedetomidine combined with dezocine in gynecological laparoscopic surgery after general anesthesia
Baiqiang PAN ; Wenjing LU ; Jie LIN ; Long JIANG ; Suya LIU
Chinese Journal of Primary Medicine and Pharmacy 2017;24(16):2490-2494
Objective To observe the adverse reaction of dexmedetomidine(Dex)combined with dezocine in gynecological laparoscopic surgery after general anesthesia.Methods 120 cases of gynecologic laparoscopic surgery,ASA Ⅰ-Ⅱ,according to random number table method were divided into three groups,40 cases in each group.A group was intravenously injected Dex 1μg/kg 10min before induction,intravenously injected dezocine 5mg+ ondansetron 4mg 30min before the end of the operation.B group was intravenously injected dezocine 5mg+ ondansetron 4mg 30min before the end of surgery.C group was intravenously injected saline 1mL+ ondansetron 4mg.Before induction(T0),5min before extubation(T1),extubation(T2),10min after extubation(T3),the MAP,HR,SpO2 of three groups were recorded.The postoperative recovery time,degree of agitation,extubation and Ramsay sedation score,VRS analgesia 24h score,postoperative adverse reaction were compared among three groups.Results There was no difference in the recovery time among the three groups(t=0.94,0.97,1.02,all P>0.05).Compared with T0,the MAP of the three groups at the time of T2 increased,but the MAP of B group and C group was significantly higher(t=4.65,5.201,all P<0.01),HR was significantly higher(t=2.382,2.915,all P<0.05).Compared with A group and B group,the agitation rate of C group was higher(u=5.54,3.47,all P<0.01).The ramsay sedation score of 1 in A group was lower than C group,the VRS score of painless rate in A,B group was higher than that of C group(all P<0.05).Postoperative 24h,there were 19,23,24 cases of the three groups with nausea and vomiting,and the incidence rates were 47.5%,57.5%,60%,respectively,the differences were not statistically significant among the three groups(all P>0.05).Conclusion General anesthesia in gynecological laparoscopic surgery using Dex combined with dezocine can make patients more stable circulation,improve patients'' recovery quatity,it is a safe and effective method for prevention of adverse reaction after general anesthesia.
5.Pondering on the Acceleration of High-Tech Medicine Industry Development
Rui PAN ; Hui HUANG ; Jie WANG ; Jianhua LU ; Jianguo WU
China Pharmacy 2001;0(07):-
OBLECTIVE:To promote a sustained and quick development of the high-tech medicine industry in our country.METHODS:The related literature was consulted and the current developmental status of the high-tech medicine in-dustry in our country was analyzed.RESULTS&CONCLUSIONS:The high-tech industry in our country can be pushed on only through rearrange industry mix at the right time,constructing a serial comprehensive development terrace,supporting es-pecially those medicine enterprises that with core technology and independent intellectual property rights,building a virtual strategic league together with the academy of higher learning and scientific research institutes.
6.Detection of early brain injury in a piglet model after deep hypothermic circulatory arrest with magnetic resonance imaging
Haipeng ZHAO ; Jie LU ; Yue TANG ; Jun ZHENG ; Xudong PAN ; Lizhong SUN
Chinese Journal of Thoracic and Cardiovascular Surgery 2013;29(8):470-474
Objective Optimal detection of deep hypothermic circulatory arrest (DHCA)-induced early brain injury is important but the effective technique is still not available in the present.The relationship between diffusion wcighted imaging (DW1) and histopathological changes in DHCA-induced piglet brain injury model were analyzed.Methods Eighteen pigs underwent deep hypothermic circulatory arrest and were divided into three groups:group A (n =6) served as control,only underwent anesthesia and thoracotomy,without extracorporeal circulation.Group B (n =6) served as underwent deep hypothermic circulatory arrest at 20 ℃ for 120 min,survived for 1 day.Group C (n =6) served as underwent deep hypothermic circulatory arrest at 20 ℃ for 120 min,survived for 2 days.Diffusion-weighted imaging and histopathology were used to study the brain injury.Results There were hematoxylin-eosin (+) and terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (+) cells in cerebral cortex and hippocampus.Hyperintensity was most evident in the cerebral cortex in group B and group C with diffusion-weighted imaging,but it was not seen in hippocampus in both groups.There were 5 and 6 piglets can be seen hyperintensity with diffusion-weighted imaging sequence in group B and group C respectively.Hyperintensity was also seen in T2WI sequence in 3 and 5 piglets in group B and group C respectively.Conclusion The study demonstrates the feasibility of diffusion-weighted imaging on evaluation of brain injury after deep hypothermic circulatory arrest and cerebral cortex was selectively vulnerable to cell injury.It is superior to conventional imaging.
7.Study on Compatibility of TCM Prescriptions for Treating Alzheimer Disease Based on Data ;Mining
Sen LIN ; Jie LU ; Kun SONG ; Zongfu CHEN ; Sen JIANG ; Chunliang PAN ; Xuebo LI ; Xianjun FU
Chinese Journal of Information on Traditional Chinese Medicine 2015;(5):41-44
Objective To study the compatibility of TCM prescriptions of TCM practitioners of all dynasties of Alzheimer disease (AD). Methods Amnesia, forgetting, dementia, and idiot were set as search words to retrieve relevant literature in Encyclopadia of Traditional Chinese Medicine. Prescription information was screened and standardized to build database. Frequency analysis and association rules were used to mine TCM prescriptions and compatibility rules. Results Totally 449 AD related prescriptions were selected, involving 682 Chinese medicinal herbs. The individual herb with the highest frequency was Ginseng Radix Rhizoma (192);the herbal pair with the highest frequency was Ginseng Radix Rhizoma-Polyhalae Radix (182);the herbal combination with 3 Chinese medicinal herbs with the highest frequency was Poria with Hostwood-Ginseng Radix Rhizoma-Polyhalae Radix (79);the herbal combination with 4 Chinese medicinal herbs with the highest frequency was Polyhalae Radix-Ginseng Radix Rhizoma-Poria with Hostwood-Glycyrrhizae Radix et Rhizoma (37). The results of association rules showed that Ginseng Radix Rhizoma-Polyhalae Radix, Ginseng Radix Rhizoma-Glycyrrhizae Radix et Rhizoma, and Ginseng Radix Rhizoma-Poria with Hostwood were commonly used compatibilities in AD related prescriptions. Conclusion Treatment of TCM practitioners in all dynasties for AD mainly chooses Chinese medicinal herbs with the efficacy of tonifying qi and soothing nerves. The compatibilities and combinations are reasonable and with certain representativeness.
9.Changes of portal pressure before and after interventional embolization with α-cyanoacrylate alkyl for portal hypertension and their clinical significance
Junyan DU ; Wei WU ; Lijie LU ; Chengan LI ; Li SONG ; Liming ZHU ; Jie PAN
Chinese Journal of Digestion 2009;29(2):82-85
Objective To study the changes of portal vein pressure before and after interventional embolization with α-cyanoacrylate alkyl and their clinical significance. Methods Thirty five patients with esophageal-gastric variceal bleeding were included. Esophageal-gastric fundal varices were embolized with e-cyanoacrylate alkyl in 35 patients, and 10 of them were treated with combination of partial splenic emblization. Portal pressure was measured directly before and after interventional embolization. The patients were followed up for 3-18 months. The changes of the liver functions and complications after the embolization were observed. Results All bleeding were controlled successfully. The main portal pressure in those who received variceal embolization only increased significantly from (32.54±5.23) cm H2O (1 cm H2O=0. 098 kPa) to (37.45±5. 11) cm H2O; superior mesenteric vein pressure increased from (31.46±4.35) cmH2O to (34.33±4.68)cm H2O; and the level of serum albumin raised from (30.45±5.78) g/L to (34.57±6.84) g/L 3 weeks after embolization. Whereas the main portal pressure in those who received both variceal and partial splenic embolizations, decreased significantly from (32. 68±4. 89) cm H2O to (28. 70±4. 58) cm H2O; superior mesenteric vein pressure decreased from (31.46±4.35) cm H2O to (28.03±4.12) cm H2O; and splenic vein pressure decreased from (32.89±4.79) cm H2O to (28.81±5. 12) cm H2O.Conclusions Esophageal-gastric variceal embolization is effective for varieeal bleeding. The increase of main portal pressure after embolization may be benefit for liver function, while the rise of superior mesenteric vein pressure may increase the risk of portal hypertensive gastrointestinal re-bleeding.Esophageal-gastric variceal embolization combined with partial splenic embolization may reduce the incidence of portal hypertension and gastrointestinal re-bleeding, but will increase the risk of ascites.
10.Effective expression and immunogenicity analysis of HIV-1 HXB2 subtype Tat protein deleted the cysteine-rich region in E. coli
Lu CHEN ; Songhua DENG ; Jie CAO ; Jun HE ; Qiuli CHEN ; Shaohua JIANG ; Wenting LIAO ; Wei PAN
Chinese Journal of Microbiology and Immunology 2008;28(5):404-410
Objective Deleting the cysteine-rich region (22-37 amino acids)of HIV-1 HXB2 Tat protein(whole length is 101 amino acids) to improve its stability and expression level in E.coli and to analyze the immunogenicity of Tat protein without the cystein-rich region [Tat(△C)protein]. Methods Tat DNA deleted the cysteine-rich region (64-111 nucleotides), named as Tat(△C)DNA, was obtained in vitro by PCR and cloned into pET-32a vector. pET-32a-Tat(△C)plasmid and the pET-32a-Tat plasmid were established and transformed into E.coli BL21(DE3) strains respectively to express and purify the protein. Three rabbits were vaccinated with pET-32a-Tat(△C)protein, then testify the reactivity of sera from rabbits by ELISA and Western blot. Results The dense of the purified pET-32a-Tat(△C)protein was 7.12 mg/ml,which was greatly more than pET-32a-Tat protein(1.50 mg/ml). Dimer of pET-32a-Tat protein can be observed just after the protein purification and stored at 25℃ and 4℃ for 7 days, but dimer of pET-32a-Tat(△C)protein was not formed at the same condition. Experimental rabbits were immunized with pET-32a-Tat(△C)protein and produced high titre of anti-pET-32a-Tat(△C)serum(1∶320 000), the antibody can react specifically with Tat(△C)protein, Tat protein (1-101 AA)and synthetic Tat(1-86 AA) protein. Deletion mutation of the cysteine-rich region of Tat protein was first performed in the study. Conclusion The expression level in E.coli and the stability of Tat protein deleted the cysteine-rich region can be increased greatly, and the protein remains good immunogenicity. The results may provide a novel antigen for further development of HIV-1 Tat vaccine.