1.Effect of ulinastatin on oxidative stress injury to myocardial ceils in diabetic rats in vitro
Huibi OUYANG ; Weifeng TU ; Ling SU ; Xi CHEN ; Wenbin QIE
Chinese Journal of Anesthesiology 2015;35(3):303-306
Objective To evaluate the effect of ulinastatin on oxidative stress injury to myocardial cells in diabetic rats in vitro.Methods The H9c2 cells were cultured in DMEM culture medium and the cells at the logarithmic growth phase were seeded in 96-well plates (density 1 × 104 cells/ml,200 μl/well) or in 6-well plates (density 1× 105 cells/m1,2 ml/well).The cells were randomly divided into 4 groups (n=18 each) using a random number table:normal control group (group C),high-glucose group (group HG),high-glucose + oxidative stress group (group HG+OS),ulinastatin +high-glucose+oxidative stress group (group U+HG+OS).The cells were cultured in high-glucose DMEM culture medium (25.0 mmol/L) for 48 h in group HG.After the cells were cultured in high-glucose DMEM culture medium for 24 h,H2O2 with the final concentration of 500 μmol/L was added to the high-glucose culture medium,and the cells were continuously cultured for 24 h in HG+OS and U+HG+OS groups.In group U+HG+OS,ulinastatin 400 U/ml was added to the high-glucose culture medium.The cells were collected for determination of cell viability,H9c2 apoptosis,activity of superoxide dismutase (SOD) and contents of malonadehyde (MDA).Apoptosis rate was calculated.The cell culture supernatant was collected for detection of lactate dehydrogenase (LDH) activity.Results Compared with group C,the cell viability and SOD activity were significantly decreased,and the apoptosis rate,MDA content and LDH activity were increased in the other groups.Compared with HG group,the cell viability and SOD activity were significantly decreased,and the apoptosis rate,MDA content and LDH activity were increased in HG+OS and U+HG+OS groups.Compared with group HG+OS,the cell viability and SOD activity were significantly increased,and the apoptosis rate,MDA content and LDH activity were decreased in group U + HG+ OS.Conclusion Ulinastatin can mitigate oxidative stress injury to myocardial cells in diabetic rats,and inhibited cell apoptosis is involved in the mechanism.
2.Effect of pretreatment with dexmedetomidine alone or in combination with sufentanil on myocardial ischemia-reperfusion injury in rats
Ling SU ; Weifeng TU ; Xi CHEN ; Wenbin QIE ; Huibi OUYANG
Chinese Journal of Anesthesiology 2013;33(5):622-625
Objective To investigate the effect of pretreatment with dexmedetomidine alone or in combination with sufentanil on myocardial ischemia-reperfusion (I/R) injury in rats.Methods Fifty healthy male Sprague-Dawley rats,weighing 250-300 g,were anesthetized with intraperitoneal pentobarbital sodium 60 mg/kg.Myocardial I/R was induced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 120 min of reperfusion.The rats were then randomly divided into 5 groups (n =10 each):sham operation group (group S),group I/R,dexmedetomidine pretreatment group (group DP),sufentanil pretreatment group (group SP),and dexmedetomidine + sufentanil pretreatment group (group DS).In group S the anterior descending branch was only exposed but not ligated.Dexmedetomidine 0.5μg/kg and sufentanil 0.1μg/kg were injected intraperitoneally 30 min before ischemia in groups DP and SP,respectively.Dexmedetomidine 0.5 μg/kg and sufentanil 0.1 μg/kg were injected intraperitoneally 30 min bbefore ischemia in group DS.Arterial blood samples were collected at 120 min of reperfusion for determination of serum creatine kinase (CK) and lactic dehydrogenase (LDH)concentrations.The rats were sacrificed at 120 min of reperfusion and hearts were removed for microscopic examination.Myocardial infarct size was calculated.The malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in myocardial tissues were measured.Results Compared with group S,the serum CK and LDH concentrations were significantly increased,the myocardial infarct size was enlarged,and SOD activity was decreased in the other groups,MDA content was significantly increased in groups I/R,DP and SP (P < 0.05 or 0.01).Compared with group I/R,the serum CK and LDH concentrations,MDA content and myocardial infarct size were significantly decreased,and SOD activity was increased in groups DP,SP and DS (P < 0.05).Compared with group DS,the serum CK concentration was significantly increased,the myocardial infarct size was enlarged,and MDA content was increased in groups DP and SP,and LDH concentration was significantly increased and SOD activity was decreased in group DP (P < 0.05).The pathological changes were significantly attenuated in groups DP and SP compared with group DS.Conclusion Dexmedetomidine pretreatment can reduce myocardial I/ R injury in rats,dexmedetomidine combined with sufentanil pretreatment provides better efficacy than either alone,and inhibition of lipid peroxidation is involved in the mechanism.
3.Clinical significance of BRCA1 and Ki-67 expression in breast cancer
Jingjing WU ; Xi CHEN ; Xiong CHEN ; Xuenong OUYANG ; Xiaolong WU ; Yan ZHANG
Cancer Research and Clinic 2014;26(1):1-5
Objective To investigate the clinicopathologic features and prognostic significance of BRCA1 and Ki-67 expression in breast cancer.Methods The expression levels of BRCA1 and Ki-67 were assayed by immunohistochemistry in 194 cases of breast cancer tissues.The correlations of BRCA1 and Ki-67 expression with patients' clinicopathologic features were also analysed.Results Low expression of BRCA1 was detected in the lymph node metastasis group,ER/PR negative group,and HER-2 positive group in 194 patients with breast cancer (P < 0.05),as well as in the triple negative breast cancer (TNBC) group compared to non-TNBC group (P <0.05).High expression of Ki-67 was detected in patients with higher histological grade,negative ER/PR,and positive HER-2 (P < 0.05).Furthermore,negative correlation was found between the expression of BRCA1 and Ki-67 (P < 0.05).The combination of low expression of BRCAl and high expression of Ki-67 was mostly found in the patients with postmenopausal,lower histological grade,lymph node metastasis,negative ER/PR and positive HER-2 (P < 0.05).However,there was no significant difference between TNBC and non-TNBC.Conclusion Joint detection of BRCA1 and Ki-67 might play an important role in predicting clinical outcomes of breast cancer,especially BRCA1 may be one of prognostic factors in TNBC.
4.Influence of Chinese herbal medicine Feitai Capsule on completion or delay of chemotherapy in patients with stage IIIB/IV non-small-cell lung cancer: a randomized controlled trial.
Shuiqiu DENG ; Xuenong OUYANG ; Zongyang YU ; Xihu DAI ; Xi CHEN ; Fangzheng FANG ; Wenwu WANG ; Zhizhen LIU
Journal of Integrative Medicine 2012;10(6):635-40
Chemotherapy completion rate can reflect the tolerance and compliance of patients to chemotherapy. Poor tolerance may result in delay or suspension of the comprehensive treatment plan, thus affect the efficacy of cancer treatment. Evaluating methods to improve the completion rate of chemotherapy and reduce the occurrence of delayed chemotherapy has gained increasing attention and is the significant area of study in the field of cancer treatment. Studies have shown that Chinese medicine combined with chemotherapy could improve the quality of life in patients with stage IIIB/IV non-small-cell lung cancer (NSCLC).
5.Rapid detection of extrinsic harmful contaminants using quantum dots-coated probes and their application prospects in traditional Chinese medicine.
Xi-Hui YANG ; Wei-Jun KONG ; Mei-Hua YANG ; Shi-Lin CHEN ; Ming ZHAO ; Zhen OUYANG
China Journal of Chinese Materia Medica 2013;38(23):3997-4005
The contamination of extrinsic harmful contaminants including mycotoxins, heavy metals and pesticides, etc, brings serious risks to traditional Chinese medicines (TCMs), further to human health. Due to their unique photoluminescence, chemiluminescence, electrochemical and electrochemiluminescence properties, semiconductor quantum dots (QDs) nanoparticles are widely used to immobilize bioprobes and biosensors, etc. In this review, the luminescence characteristics and specific ligands of QDs probles which are used to determine contaminants were summed up. Then, the applications of QDs-coated novel probes in the determination of mycotoxins, heavy metals and pesticides were discussed in detail. In addition, the contamination levels and characteristics of extrinsic harmful residues in TCMs were investigated. Further, the maximum levels of those contaminants in TCMs were compared with those set by various countries. Finally, the future development trends and problems of QDs-coated probes in the determination of those extrinsic residues in TCMs were prospected.
Drug Contamination
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prevention & control
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Humans
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Medicine, Chinese Traditional
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methods
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Nanotechnology
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instrumentation
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methods
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Quantum Dots
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Safety
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Time Factors
6.Prokaryotic expression and purification of anti-AEG-1 single-chain variable antibody
Man LU ; Xinyang LIU ; Min LONG ; Chen WANG ; Chong LIU ; Zhaowei GAO ; Yongri OUYANG ; Xi CHEN ; Huizhong ZHANG
International Journal of Laboratory Medicine 2015;(1):20-21,24
Objective To construct anti-astrocyte elevated gene-1(AEG-1)single-chain variable antibody (V23)prokaryotic ex-pression vector,and to conduct the protein purification and immunological activity detection.Methods The Primer5 software was applied to design the primers aiming at the gene sequence of the antibody anti-AEG-1 single-chain variable region for constructing the prokaryotic expression plasmid of PRsetC/V23.After the enzyme digestion by the restriction enzyme Pst1 and correct DNA se-quencing,the prokaryotic expression plasmid was led to E.coli BL21 ,the prokaryotic expression engineering strain containing the V23 gene was constructed.After the induction with IPTG,the interest protein was purified by the magnetic beads with the HIS tag,and the content of interest protein was determined by the SDS-PAGE electrophoresis.Western blot and ELISA were adopted to detect the immune activity of the nti-AEG-1 single-chain variable region antibody.Results For the constructed prokaryotic expres-sion plasmid PRsetC/V23,the single enzyme digestion and sequencing analysis displayed that the constructed V23 gene was com-pletely consistent to the designing sequences.After IPTG induction,SDS-PAGE electrophoresis showed an apparent protein band at 31×103 ,the Western blot detection showed a specific AEG-1 response band at 80 ×103 ,the ELISA test showed the positive re-sults.Conclusion The PRsetC/V23 prokaryotic expression plasmid and the V23 prokaryotic expression engineering strain are suc-cessfully constructed,this engineering strain can express anti-AEG-1 single-chain variable region antibody protein,and the protein has good immune activity.
7.Relationship between sCD44v6 expression and TCM differentiation type of gastric carcinoma patients and influence of weitai capsule on the expression.
Xi CHEN ; Xue-nong OUYANG ; Xi-hu DAI ; Mei-hua CHEN ; Qiu-cheng LIN
Chinese Journal of Integrated Traditional and Western Medicine 2005;25(1):12-15
OBJECTIVETo explore the relationship of TCM type with serum level of soluble CD44v6 (sCD44v6) and different histologic parameters in gastric carcinoma patients and to observe the influence of Weitai capsule (WTC) on the sCD44v6 expression.
METHODSTCM typing and sCD44v6 expression were determined in all the enrolled patients (30 in the control and 32 in the trial group) before operation, and 3-4 courses of chemotherapy was applied to them from 3-4 weeks after operation. To the patients of trial group, oral administration of WTC was given additionally with 4 capsules, 3 times a day for consecutive 3 months.
RESULTSsCD44v6 was significantly positive correlated with the degree of cancer cell differentiation, infiltration and lymph node metastasis; (2) Level of sCD44v6 was the highest in patients of blood stasis type, as compared with that in the patients of Pi-deficiency type or of damp-heat type, the difference was significant; (3) After ending treatment, level of sCD44v6 in the trial group was significantly lower than that in the control group.
CONCLUSION(1) Serum level of sCD44v6 could be taken as the criterion for evaluating the development and prognosis of gastric cancer, as well as the therapeutic target for anti-metastasis treatment; (2) Serum level of sCD44v6 is related to some extent with TCM type of blood stasis and Pi-deficiency; (3) WTC combined with chemotherapy could further inhibit the expression of serum sCD44v6 in gastric carcinoma patients.
Adenocarcinoma ; drug therapy ; immunology ; Adult ; Aged ; Antineoplastic Agents, Phytogenic ; therapeutic use ; Capsules ; Cell Adhesion ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Glycoproteins ; blood ; Humans ; Hyaluronan Receptors ; blood ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy ; Stomach Neoplasms ; drug therapy ; immunology
8.Effects of perindopril on expression of kidney aquaporin-2 and urine aquaporin-2 excretion in chronic heart failure rats.
Shao OUYANG ; Wei CHEN ; Xi-Bin KUANG
Chinese Journal of Cardiology 2013;41(4):276-281
OBJECTIVETo determine the expression of kidney aquaporin-2 (AQP2) and urine AQP2 excretion in chronic heart failure (CHF) rats and investigate effects of perindopril on the expression and excretion of AQP2.
METHODSSixty rats were randomized into three groups: control group, CHF group, CHF + Perindopril group. According to left ventricular myocardial infarction size, CHF group and perindopril group were further divided into heart failure subgroup (LVMI ≥ 20%) and cardiac functional compensation subgroup (LVMI < 20%), respectively. Blood and urine samples were collected from the rats for measuring serum Na(+), urine volume and urine osmolality. The concentration of plasma arginine vasopressin (p-AVP) was detected by radioimmunoassay (RIA). Immunohistochemistry, semi-quantitative real time-polymerase chain reaction (RT-PCR) and Western blot were performed for measurement of kidney inner medullary AQP2. The concentration of Urine AQP2 was measured by indirect enzyme-linked immunosorbent assay (indirect ELISA).
RESULTSImmunohistochemistry, RT-PCR, Western blot examinations revealed increased quantity of the inner kidney medullary AQP2 expression (0.2013 ± 0.0417), AQP2 mRNA (0.98 ± 0.33) and AQP2 protein expression (0.94 ± 0.21) in heart failure subgroup (n = 13) compared to control group (n = 20, 0.1518 ± 0.0214, 0.58 ± 0.51, 0.51 ± 0.46), which could be significantly by perindopril (n = 13, 0.0712 ± 0.0218, 0.76 ± 0.45, 0.82 ± 0.49, all P < 0.05 vs. heart failure subgroup). The concentration of plasma arginine AVP [(19.72 ± 3.91) ng/ml] and Urine AQP2 [(82.52 ± 11.77) ng/L] were significantly higher in heart failure subgroup than in control group [n = 20, (51.67 ± 12.58) ng/L, (6.94 ± 3.10) ng/ml] (P < 0.05), which were significantly reduced by perindopril [n = 13, (15.65 ± 4.10) ng/L, (71.65 ± 9.21) ng/ml].
CONCLUSIONIncreased expression of the kidney inner medullary AQP2 and the excretion of urine AQP2 in chronic heart failure rats could be reduced by perindopril.
Animals ; Aquaporin 2 ; metabolism ; urine ; Disease Models, Animal ; Heart Failure ; metabolism ; Kidney ; drug effects ; metabolism ; Male ; Perindopril ; pharmacology ; Rats ; Rats, Sprague-Dawley
9.A novel technique to preserve the alveolar ridge width following tooth extraction in the maxillary frontal area
Xi JIANG ; Ye LIN ; Yu ZHANG ; Ping DI ; Bo CHEN ; Xiulian HU ; Jia LUO ; Shuxin REN ; Siyuan OUYANG
Journal of Peking University(Health Sciences) 2016;48(1):175-179
Objective:To introduce a novel ridge preservation technique with micro-titanium plate avoiding the use of bone grafting materials,and evaluate the potential horizontal bone preservation effect of this new technique,applied on single maxillary central incisors after tooth extraction for future implant restoration.Methods:Nine patients (six women and three men),mean age (26.0 ±5.7)years(from 1 8 to 34 years)referred to the Department of Oral Implantology,Peking University School and Hospital of Stomotology,were selected and diagnosed with unsalvageable single middle incisor with fine general con-ditions,no signs of acute local inflammation,no ongoing or previous periodontitis,healthy neighboring teeth and intact buccal bone walls.Tooth extraction,delayed implant placement and implant-supported single crown restoration were selected as treatment plan.The teeth were extracted atraumatically with lo-cal anesthesia,followed by a reflection of a minor flap to exposed 2-3 mm of the buccal bone plate.Af-ter that,a micro-titanium plate was trimmed and bended to fit the convexity of the labial bone and fixed by two mini pins with intent to support the labial soft tissue.The flap was then repositioned over the micro titanium plate and secured with two single sutures.No bone grafting materials or releasing incisions were needed.The sockets were left to heal without any intention of primary wound closure.Cone-beam compu-ted tomographic (CBCT)scans were obtained before and four months after tooth extraction.Horizontal ridge widths were measured with CBCT software,and the preservation effects were calculated and recor-ded by the percentage of horizontal ridge alteration.Results:The nine extraction sockets were healed un-eventfully.The average socket width before extraction was (7.51 ±0.48)mm (6.92-7.82 mm).The average alveolar ridge labial-palatal width at the control point of the edentulous area was (6.81 ±0.44) mm (6.04-7.38 mm)4 months after tooth extraction,the mean percentage of ridge width preserved was 90.87%±2.91%(87.28%-95.60%).Conclusion:This novel ridge preservation method by the usage of a micro-titanium plate did not interfere with the natural socket healing process,and at the same time,largely preserved the width of alveolar ridge without any bone grafting procedures.Long term results remain to be seen.
10.The value of transrectal ultrasound and magnetic resonance imaging fusion targeted prostate biopsy in biopsy-naive men
Jianquan HOU ; Qilin XI ; Jinxian PU ; Chen HUANG ; Jun OUYANG ; Gang LI ; Yuhua HUANG ; Chao MA ; Zixian WANG
Chinese Journal of Urology 2017;38(6):469-472
Objective To estimate the value of transrectal ultrasound/magnetic resonance imaging (TRUS/MR) fusion targeted prostate biopsy(targeted biopsy,TB) in the biopsy naive patients.Methods Between September 2015 and September 2016,91 patients with PI-RADS ≥ 3 suspicious regions on the multiparametric magnetic resonance imaging (mpMRI) were retrospectively evaluated.The age of patients was 46-83 years (median 68).Serum PSA level before biopsy was 1.2-85 ng/ml (median 11.2 ng/ ml),in which 36 cases with PSA < 10 ng/ml,30 cases 10-20 ng/ml,and 25 cases > 20 ng/ml.Two-core TB using real-time virtual sonography (RVS) platform for mpMRI-suspicious lesions was followed by 12-core systematic biopsy (SB).The detection rates for any cancer (PCa) and clinically significant prostate cancer (CsPCa) were compared between TB and SB.Results The total detection rate for PCa was 57.1%,with a comparable positive rate between TB (44.0%) and SB (51.7%) groups which did not significantly differ (P =0.14).The proportion of CsPCa in TB group was higher than that in SB group (80.0% vs.68.1%,P =0.21).In TB group,detection of PCa for grade 5 lesions was significantly higher than that for grade 3 lesions (77.1% vs.10.3%,P <0.001).Detection of PCa was comparable between TB and SB groups in different regions of PSA < 10 ng/ml,10 ~ 20ng/ml and > 20ng/ml (27.8% vs.36.1%,50% vs.56.7%,60% vs.68%,respectively).Conclusions This study revealed a similar rate of prostate cancer detection between 2-core targeted biopsy guided by TRUS/MR fusion and 12-core random biopsy in different PSA regions for no prior biopsy men.TB maybe tend to detect high proportion of CsPCa.PI-RADS is instructive to select appropriate patients for TB.