1.Protection of quercetin against hyperbaric oxygen-induced apoptosis of human lens epithelial cell and its mechanism
Li-bin, CHANG ; Yong-zhen, BAO ; Yi, CHEN ; Wen-zhen, YU ; Xiao-xin, LI
Chinese Journal of Experimental Ophthalmology 2012;30(6):485-489
Background Oxidative stress-induced apoptosis of human lens epithelial cells (LECs) is associated with c-Jun N terminal kinase (JNK) pathway.Quercetin possesses the antioxidation by inhibiting the JNK pathway.However,whether quercetin can protect LECs from the oxygen-induced damage is still not proved.Objective This study attempted to invatigate the effects and its mechanism of quercetin against hyperbaric oxygeninduced LECs apoptosis. Methods Human LECs line SRA01/04 was cultivated and passaged in MEM medium containing 10% fetal bovine serum and 0.5% non-essential amino acids for 2 hours,with or without 20 μmol/LSP600125 or 1 μmol/L quercetin prior to exposure to hyperbaric oxygen.Each exposure session remained 6 hours in 99% O2 and 1%CO2 with a pressure chamber at 588 kPa.The viability of human LECs was detected by MTT.Cell apoptosis was assessed by flow cytometer using Annexin V-FITC apoptosis detection.The expression of JNK/p-JNK,c-Jun/p-c-Jun,caspase 3 and caspase 9 were detected by Western blot. Results LECs viability (A570 ) was 0.835 ±0.082,0.450±0.083,0.654±0.079,0.649±0.090 respectively in the blank control group,hyperbaric oxygen exposed group,hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group.The A570 in the hyperbaric oxygen exposed group was significantly lower than the blank control group ( P =0.000),but those in hyperbaric oxygen + SP600125 group and hyperbaric oxygen+quercetin group were significantly higher than the hyperbaric oxygen exposed group ( P =0.003,0.002 ).The numbers of apoptosis cells were 3.17 ±0.74,19.77 ± 1.44,8.45 ±0.93,7.79 ±0.78 respectively in the blank control group,hyperbaric oxygen exposed group,hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group.Apoptotic LECs were significantly increased in the hyperbaric oxygen exposed group compared with the blank control group ( P=0.000),but those in the hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group were significantly reduced in comparison with hyperbaric oxygen exposed group (both P=0.000).In additional,expressions of p-JNK,p-c-Jun,caspase 3 and caspase 9 proteins in the cells were elevated in the hyperbaric oxygen exposed group compared with the blank control group (all P =0.000 ),however,those in the hyperbaric oxygen + SP600125 group and hyperbaric oxygen + quercetin group were declined when compared with the hyperbaric oxygen exposed group( all P<0.05 ). Conclusions JNK pathway is involved in the apoptotic procedure of human LECs induced by oxygen stress.SP600125 and certain concentration of quercetin can interdict the JNK signal pathway and endogenous apoptosis of LECs and further alleviate hyperbaric oxygen-induced damage of LECs.
2.Selection and preliminary identification of human Fab fragement antibody against amylin from phage antibody library
Yunfeng ZHEN ; Changying LI ; Jiwu CHANG ; Tiehong ZHU
Chinese Journal of Immunology 2009;25(12):1112-1115
Objective:To obtain antibodies against amylin from a 'naive' human Fab fragment antibody phage diasplay library and to analyze the specificity of antigen binding activity.Methods:Panning and screening Fab antibody from the antibody library,the positive clones with well reactivity to amylin were selected after five times selection of 'adsorption-elution-enrichment'.Then the plasmid DNA which was extracted from the clones,was digested with Spe Ⅰ and Nhe Ⅰ to delete gⅢ (about 660 bp).The digested 47 000 bp DNA which was purified after separation of bands from agarose gel was ligated with T4-DNA ligase.The constructed expressing phagemids were transformed to the BL21(DE3)pLysS,soluble Fab was expressed in it by the induction of IPTG and its characteristics and specificity were determined by ELISA and Western blot.Results:Soluble Fab antibodies were expressed in E.coli.According with molecular weight of IgG Fab,protein band of about 47 kD was shown by SDS-PAGE.Western blot using the goat anti human IgG-HRP showed their binding activities.ELISA showed their specificity with amylin antigens and they did not react with bovine serum albumin.Conclusion:The high level expression and identification of the soluble human anti- amylin Fab fragment antibodies has been obtained successfully,which lays a solid foundation for further researching about the biological and pathological activities of amylin.
3.Research on Collateral-unblocking Medicines for Cardiovascular Diseases under the Guidance of Vessels Collateral Theory.
Cong WEI ; Li-ping CHANG ; Zhen-hua JIA
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(12):1513-1516
"Vessels Collateral Theory", as the inherit and development of "blood vessels" in Huangdi Neijing, is a systematic system by integrating severe vascular diseases. In this article, by taking cardiovascular diseases (CVD) as a cut-in point, roles of "minute collateral-microvascular" lesions in the occurrence and development of CVD were further explored. The interventional effect of collateral-unblocking medicines under the guidance of Vessels Collateral Theory was also in-depth explored, hoping to leading Chinese medical prevention and treatment of CVD.
Cardiovascular Diseases
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drug therapy
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Humans
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Medicine, Chinese Traditional
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Research
4.Total hip arthroplasty for post-traumatic hip fusion ankylosis secondary to operation in acetabular combined with femoral head fracture
Ping ZHEN ; Xusheng LI ; Qi TIAN ; Shenhu ZHOU ; Yanfeng CHANG
Chinese Journal of Orthopaedics 2016;36(19):1213-1221
Objective To investigate the technique and clinical results of total hip arthroplasty in treating post-traumatic hip fusion ankylosis secondary to operation in acetabular combined with femoral head fracture.Methods From October 2009 to January 2015,7 patients (7 hips) with post-traumatic hip fusion ankylosis underwent total hip arthroplasty.There were 6 males and 1 female with an average age of 38 years (range,25-51 years).There were 4 hips on the left side,3 hips on the right side.Open reduction and internal fixation were performed on all patients.One patient had a postoperative deep infection and 6 patients underwent implant removal.The hip bony fusion ankylosis was developed secondary to post-traumatic arthritis after the surgery of ipsilateral acetabular combined with femoral head facture.The interval between internal fixation and total hip arthroplasty was 37.6 months on average (range,21-67 months).The hip bony fusion ankylosis was relieved by wedge shape osteotomy at femoral neck level.The cemenfless prostheses were implanted in suitable places based on the accuracy position of acetabular center.One infection case was treated via surgical debridement,removal of all the screws,and antibiotic-loaded cement spacer implantation in the first stage.The cementless prostheses were implanted in the second stage.Results The average operation time and blood loss was 108 min (90-150 min) and 320 ml (280-450 ml) respectively.The internal fixation devices were explanted completely,including plate and screw in 5 cases,only screw in 1 case,intramedullary nail and screw in 1 case.The mean fallow-up period was 32.5 months (range,15-48 months).The average Harris hip score improved from 48.8±6.5 points preoperatively to 92.6±5.1 points post-operatively (t=22.82,P=0.001).Osseointegration was developed in all of the acetabular and femoral components at 3 months post-operatively.Radiograph analysis showed satisfied position of acetabular cup and no evidence of implant migration or center of rotation change.Stem subsidence (<1.0 mm) occurred in 2 cases and heterotopic ossification in 2 cases (Brooker type Ⅰ and type Ⅱ in 1 case,respectively) at one year postoperatively.No complication occurred,such as damages of vessels and nerve,dislocation,component loosening or infection.Conclusion The wedge shape osteotomy of femoral neck is a safe and reliable method in treating hip fusion ankylosis.Total hip arthroplasty can be achieved by reconstruction of acetabular center,suitable components implanted in anatomical position and ideal reconstruction of soft tissue around hip.
5.Changes of Brain Natriuretic Peptide and Atrial Natriuretic Peptide in Serum of Children with Congestive Heart Failure
hong, CHANG ; xiu-zhen, HAN ; wen-li, WANG
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To study the serum level of brain natriuretic peptide (BNP) and atrial natriuretic peptide (ANP) in patients with congestive heart failure (CHF) and assess the value of BNP. Methods The serum levels of BNP and ANP were determined by enzyme- linked immnoabsorbent assay.Cardial mdex(CI)and left ventricular election fraction(LVEF)in heart failure stage and period recouery of children with heart failare were determined by Doppler ultrasonography cardiogram. Results The BNP serum levels began to increase before heart failure (P
7.Duraplasty with Neuropatch versus autologous fascia lata for Chiari I malformation with syringomyelia: A comparative study
LIU BIN ; WANG ZHEN-YU ; LI ZHEN-DONG ; MA CHANG-CHENG ; SUN JIAN-JUN ; CHEN XIAO-DONG
Journal of Peking University(Health Sciences) 2005;37(6):629-632
Objective: To evaluate the outcome and postoperative reaction of dural substitute (Neuropatch) applying in the treatment of Chiari I malformation(CMI) associated with syringomyelia(SM). Methods:Forty patients of CMI associated with SM were operated in our department from Jul. 2002 to Jul. 2004. All patients underwent posterior cranial fossa decompression and duraplasty. They were divided into two groups, 20 patients being repaired with Neuropatch (Neuropatch group), and the others with autologous fascia lata (fascia group). There were 6 males and 14 females in Neuropatch group and 10 males and 10 females in fascia group. The operations were performed under general anesthesia via suboccipital approach and the extent of posterior cranial fossa decompression ranged from 20 cm2 (5 cm×4 cm) to 35 cm2 (5 cm×7 cm). The removal of posterior arch of atlas depended on the extent of tonsillar herniation, and the dura was opened in Y shape. The Neuropatch was cut into triangular shape, and the same sized autologous fascia lata was used in fascia group. The patches were sutured tightly to the dura matter in each group. The incision was closed layer by layer and drainage was used, if necessary. Antibiotics and hormone were routinely used. The duration of operation, postoperative fever were evaluated, the outcome of the operation was evaluated by Tator scale, and the data were analyzed with statistic software SPSS 10.0. Results: There were12 patients (60%) who suffered from postoperative fever in the Neuropatch group, and 9 patients (45%) in the fascia group(χ2=0.902,P=0.342). Seventeen patients in each group were improved postoperatively. The duration of operation, postoperative fever and antibiotics used were compared between the two groups. No significant difference was found, but the duration of postoperative fever and the time of hormone used were different. There were no postoperative infections that occurred after the follow up for 1 to 2 years, except for one patient in fascia group who developed infective granuloma and recovered later by treatment. Conclusion: Neuropatch is a useful dural substitute for the repair of dural defects in the treatment of CMI associated with syringomyelia.
8.The study of matrix metalloproteinase-9 and tumor necrosis factor-or in predicting chorioamnhionitis in patients with premature rupture of membranes
Jin-Na HUANG ; Yuan-Cheng LI ; Li-Xuan ZHEN ; Shi-Xian GUO ; Chang GAO ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(10):-
Objective To study the relation of matrix metalloproteinase-9 in serum and tumor necrosis fac- tor-?in serum and amniotic fluid in predicting ehorioamnhionitis in patients with premature rupture of membranes (PROM).Methods The levels of MMP-9 in serum and TNF-?in serum and amniotie fluid were measured by ra- dioimmunoassay and ELISA in 67 cases with premature rupture of membranes as study group and 40 cases normal full-termed pregnant women as controls group.Results(1)The levels of TNF-?in amniotie fluid and MMP-9 in serum in study group were significantly higher than those in controls group(P0.05).(2)In study group,the levels of MMP-9 of serum in0.05).Conclusions The levels of TNF-?in amniotic fluid and MMP-9 in serum were valuable clinical indices for identification of chorioamnionitis in patients with PROM.The levels of MMP-9 in serum also could assess the time of rupture of membranes and the degree of ehorioamnionitis.
9.Serum paraquat concentration detected by spectrophotometry in patients with paraquat poisoning
Chang-Bin LI ; Xin-Hua LI ; Zhen WANG ; Cheng-Hua JIANG ; Ai PENG
World Journal of Emergency Medicine 2011;2(3):179-184
BACKGROUND: Paraquat (PQ) is a world-wide used herbicide and also a type of common poison for suicide and accidental poisoning. Numerous studies have proved that the concentration of serum PQ plays an important role in prognosis. Spectrophotometry, including common spectrophotometry and second-derivative spectrophotometry, is commonly used for PQ detection in primary hospitals. So far, lack of systematic research on the reliability of the method and the correlation between clinical features of patients with PQ poisoning and the test results has restricted the clinical use of spectrophotometry. This study aimed to evaluate the reliability and value of spectrophotometry in detecting the concentration of serum PQ. METHODS: The wavelengths for detecting the concentration of serum PQ by common and second-derivative spectrophotometry were determined. Second-derivative spectrophotometry was applied to detect the concentration of serum PQ. The linear range and precision for detection of PQ concentration by this method were confirmed. The concentration of serum PQ shown by second-derivative spectrophotometry and HPLC were compared in 8 patients with PQ poisoning. Altogether 21 patients with acute poisoning 4 hours after PQ ingestion treated in the period of October 2008 to September 2010 were retrospectively reviewed. The patients were divided into higher and lower than 1.8 μg/mL groups based on their concentrations of serum PQ measured by second-derivative spectrophotometry on admission. The severity of clinical manifestations between the two groups were analyzed with Student's t test or Fisher's exact test. RESULTS: The absorption peak of 257 nm could not be found when common spectrophotometry was used to detect the PQ concentration in serum. The calibration curve in the 0.4–8.0 μg/mL range for PQ concentration shown by second-derivative spectrophotometry obeyed Beer's law with r=0.996. The average recovery rates of PQ were within a range of 95.0% to 99.5%, relative standard deviation (RSD) was within 1.35% to 5.41% (n=6), and the lower detection limit was 0.05 μg/mL. The PQ concentrations in serum of 8 patients with PQ poisoning shown by second-derivative spectrophotometry were consistent with the quantitative determinations by HPLC (r=0.995, P<0.0001). The survival rate was 22.2% in patients whose PQ concentration in serum was more than 1.8 μg/mL, and the incidences of acidosis, oliguria and pneumomediastinum in these patients were 55.6%, 55.6%and 77.8%, respectively. These clinical manifestations were different significantly from those of the patients whose PQ concentration in serum was less than 1.8 μg/mL (P<0.05). CONCLUSIONS: For common spectrophotometry, the wavelength at 257 nm was not suitable for detecting serum PQ as no absorbance was shown. Second-derivative spectrophotometry was reliable for detecting serum paraquat concentration. Serum PQ concentration detected by second-derivative spectrophotometry could be used to predict the severity of clinical manifestations of patients with PQ poisoning, and PQ content higher than 1.8 μg/mL 4 hours after ingestion could be an important predictive factor for poor prognosis.
10.Effect of Brain Wave-Biofeedback on Attention Deficit Hyperactivity Disorder
Hong-hui LI ; Ning-zhen LI ; Zhao-de XIE ; Shaoling MO ; Jing CHANG
Chinese Journal of Rehabilitation Theory and Practice 2006;12(3):188-189
ObjectiveTo explore the effect of brain wave-biofeedback on attention deficit hyperactivity disorder.Methods29 children with attention deficit hyperactivity disorder used VBFB3000 Brain Wave-Biofeedback system to control the 4~8 Hz brain wave and activate the 12~16 Hz wave twice a week.Results84.6% children primarily with attention deficit became normal,as well as 100% with hyperactivity,91.6% with mixed appearing.ConclusionBrain Wave-Biofeedback is effective on any types of attention deficit hyperactivity disorder.