1.Cloning and prokaryotic expression of a glutathione peroxidase from tick Hemaphysalis longicornis
Chaoying ZHANG ; Guowei GUO ; Zhengchao WANG ; Xiaohong HUANG
Chinese Journal of Zoonoses 2012;(9):861-868
Ticks transmit various diseases to livestock and human beings. The researches on ticks are of great importance in medical and veterinary sciences. To express a glutathione peroxidase gene (HlGPx) from Hemaphysalis longicornis in Escherichia coli (E. coli) so as to provide basis for further studies, the gene was amplified from cDNAs of H. longicornis adult ticks by PCR prompted by information from EST library. The TGA codon encoding selenocysteine in the gene was mutated into the universal genetic code for cysteine,TGC, by site-directed mutagenesis. E. coli was transformed with the recombinant expression vector pGEX-4T-1/HlGPx'(the mutated HlGPx) and induced to express recombinant HlGPx', which was then analyzed by sodium dodecylsulphate-polyacrylamide gel electrophoresis and Western blotting. The result showed that the complete coding sequence of HlGPx was obtained successfully, the deduced polypeptide was 223 aa, with a calculated molecular mass of 25.0 kDa; the recombinant fusion protein was approximately 51 kDa, correspondent to the calculated. The antibody against glutathione S-transferase (GST) recognized the recombinant protein fused with GST in a Western blotting assay, which confirmed the result above-mentioned. In conclusion, the mutated HlGPx was expressed in E. coli successfully and it could be used for further preparation of immune serum and functional analysis.
2.Effects of ulinastatin on expression of intestinal defensin-5 mRNA in the rat model of sepsis
Longyuan JIANG ; Meng ZHANG ; Jianxing CHANG ; Zhengfei YANG ; Zhengchao LUO
Chinese Journal of Emergency Medicine 2008;17(8):848-851
Objective To investigate the effeets of ulimstatin on expression of intestinal defemin-5 mRNAin the rat model of sepsis.Method The experiment was performed in pharmaco-laboratory of medical college,Sun Yat-Sen University.sixty Sprague-Dawley rals were randomly divided into control,sepsis,pretreated andtreated groups(n=15).Semis was induced in the mts of latter three groups by cecal lifo.and puncture(CLP).The rats of pretreated group received 25 000 U/kg ulinastatin 2 hours before operation and the rats of uli-nastatin treated groups received 50 000 U/kg ulinastatin 2 hours after operation.Some pieces of ileum mucosa weretaken 12 h after CLP.Tge pathological changes were observed and the expression of RD-5 mRNA was detectedwith RT-PCR.All data were managed by SPSS 13.0 software and arIaIyzed by using One-way ANOVA and LSD-ttest.Results The expression of RD-5 mRNA in the rats of sepsis group significantly decreased compared to col-trol(P<0.05).The expression of RD-5 mRNA of pretreated and treated groups sigificantly inereased comparedto sepsis group(P<0.05);pretreated groups had more increased expression of,RD-5 mRNA compared to treatedgroups(P<0.05).Conclusions The expression of intestinal RD-5 mRNA significantly decreases in sepsis,which could be improved by the treatment of ulinadtatin leading to intestinal mucosal protection of the siqnifleant.The pretreatment may be more effective than the theTapeatic treatment in the rat model of sepsis.
3.Chemical constituents of endophytic fungus Nodulisporium sp. A4 from Aquilaria sinensis.
Dongli LI ; Zhengchao WU ; Yuchan CHEN ; Meihua TAO ; Weimin ZHANG
China Journal of Chinese Materia Medica 2011;36(23):3276-3280
OBJECTIVETo investigate the chemical constituents of an endophytic fungus, Nodulisporium sp. A4, from the medicinal plant Aquilaria sinensis and search for antitumor natural products.
METHODThe fungus was cultured in liquid medium and extracted with EtOAc. The compounds were isolated by various chromatographic methods (silica gel, reverse silica gel, Sephadex-LH20, preparative TLC and so on) and recrystallization. Structural elucidation was conducted by extensive analysis of spectroscopic data as well as by comparison with literature reports. The antitumor activity of isolated compounds was tested by MTT method in vitro.
RESULTSeven compounds were isolated and identified from the broth culture, their structures were determined to be 5-methyl-2-vinyltetrahydrofuran-3-ol (1), 6-methyl-2-(5-methyl-5-vinyltetrahydrofuran-2-yl) hept-5-en-2-ol (2), 6alpha-hydroxycyclonerolidol (3), rel-(1S,4S, 5R,7R,10R)-10-desmethyl-1-methyl-11-eudesmene (4), tyrosol (5), 8-methoxynaphthalen-1-ol (6), and 1,8-dimethoxynaphthalene (7). Three compounds were isolated and identified from the mycelia as ergosterol (8), ergosterol peroxide (9), and cerevisterol (10). The in vitro pharmalogical evaluation results displayed that compounds 3 and 4 showed 89.1%, 44.2% and 82.3%, 79.8% inhibition against tumor cell lines SF268 and NCI-H460 at 100 mg x L(-1), respectively.
CONCLUSIONCompound 1 was a new natural product, compounds 2, 3, 7 and 10 were reported from the genus Nodulisporium sp. for the first time. Compounds 3 and 4 exhibited weak inhibitory effects on the proliferation of tumor cell lines SF268 and NCI-H460.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Endophytes ; chemistry ; Humans ; Organic Chemicals ; chemistry ; isolation & purification ; pharmacology ; Thymelaeaceae ; microbiology ; Xylariales ; chemistry
4.Morphometric and functional alterations of amygdale and hippocampus in patients with depression: a MRI study
Dongqing WANG ; Yuefeng LI ; Yifeng LUO ; Qin WU ; Ruigen YIN ; Liang ZHANG ; Yunfei XU ; Liangju SHENG ; Tian ZHAO ; Wen LIU ; Zhengchao WANG ; Weibin SUN
Chinese Journal of Radiology 2011;45(7):623-627
Objective To explore the morphometric and functional alterations of amygdale and hippocampus in patients with depression by anatomical and functional MRI, and try to reveal the pattern and pathogenesis of the changes in depression. Methods Sixty patients (divided equally into mild, moderate and major groups according to patient′s scores of HAMD) and 20 healthy control groups were scanned using T1WI and fMRI. The outlines of hippocampus and amygdale were drawn manually by observer and the volumes were calculated and normalized subsequently. Functional MRI was processed using SPM5 and individual activation map was got subsequently. Dunnett-t test and Pearson correlation analysis were separately used to analyze the morphometric and functional changes and the correlations between cerebral changes and clinical severity. Results The hippocampal volumes of control groups were 2296±202 left for left side and 2283±199 for right side. The hippocampal volumes of depressive patients were smaller than those of control groups, especially for the major group (left 1978±176,Dunnett-t=-10.0,P<0.01,right 1981±171,Dunnett-t=-9.2,P<0.01). The moderate group showed moderate reduced volume(left 2127±180,Dunnett-t=-3.0,P<0.05,right 2135±183,Dunnett-t=-3.0,P<0.05), while the mild ones showed slightly decreased volume (left 2207±189,Dunnett-t=-1.4,P>0.05,right 2210±191,Dunnett-t=-1.6,P>0.05). The amygdale′s volumes of control groups was 1762±185,the right was 1749±182, while those in patient group reduced along with the patient′s condition, i.e., the mild groups (left 1992±200,Dunnett-t=4.8,P<0.01,right 1989±191, Dunnett-t=5.0,P<0.001), the moderate groups (left 1889±192, Dunnett-t=2.8,P<0.05,right 1896±195,Dunnett-t=2.8,P<0.05), and the major groups (left 1539±178,Dunnett-t=-6.8,P<0.01,right 1543±180,Dunnett-t=-7.0,P<0.01).For fMRI study, patient group demonstrated more activation of the amygdale and hippocampus under the stimulations of negative images than controls. Furthermore, the strengthens of activation decreased along with the patient′s condition, i.e., the major ones showed the weakest activation among the patients, though it was higher than that of control group. In patient group, both the volumes and activations of hippocampus and amygdale showed significant negative correlations with HAMD scores(r=-0.80--0.90,P<0.05). Conclusion The hippocampal volumes of depressive patients reduced, which may be due to the change of the amygdale, and the amygdale′s volumes were changed along with the patient′s condition. There were more activation in the amygdale and hippocampus of depressive patients under the stimulations of negative images.
5.Comparison of transperitoneal robot assisted and retroperitoneal laparascopic Anderson-Hynes dismembered pyeloplasty for the treatment of areteropelvic junction obstruction
Zhengchao FAN ; Jinshan LU ; Jie ZHU ; Wei WANG ; Xin MA ; Yong XU ; Zhifei HU ; Liang CUI ; Jinkai DONG ; Jiangping GAO ; Xu ZHANG
Chinese Journal of Urology 2012;33(6):417-420
Objective To compare the clinical effectiveness and safety of transperitoneal robot assisted and retroperitoneal laparoscopic Anderson-Hynes dismembered pyeloplasty for the treatment of ureteropelvic junction obstruction. Methods From September 2008 to June 2009,six patients with primary UPJO underwent transperitoneal robot assisted dismembered pyeloplasty (TRADP) (5 males and 1 female;average age 25 yrs,range from 14-40 yrs),of whom 4 with severe hydronephrosis,2 with intermediate.According to the demographic and preoperative information,each patient in the TRADP group was matched to two corresponding patients with primary UPJO accepting retroperitoneal laparoscopic Anderson-Hynes dismembered pyeloplasty (RLADP) in the same period.The operative time,the intracorporeal suturing time,intraoperative blood loss,the duration of the urethral catheter and the drainage time,the postoperative hospital stay and the postoperative result were compared between the 2 groups.The two groups were identical with regard to gender,side of UPJO,and surgical procedure.The mean age and BMI were comparable between the TRADP and RLADP. Results Between the two groups,the operative time was ( 157 ± 20) min vs ( 127 ± 18) min ( P > 0.05 ),the intracorporeal suturing time was (44 ± 6) min vs (49 ± 6 ) min ( P >0.05).In TRADP and RLADP groups,the intraoperative blood loss was (23 ± 8) ml vs (21 ± 17) ml ( P > 0.05 ),the duration of the drain was (47 ± 10) h vs ( 161 ± 41 ) h ( P < 0.01 ),the duration of the urethral catheter was (92 ±46) h vs ( 175 ±26) h (P <0.05),the postoperative hospital stays were (6.0 ± 0.8 ) d vs (8.0 ± 0.5) d ( P < 0.01 ).The operation was successful in all cases of two groups,with no conversion to open surgery.The follow-up of 6 -32 months,with average of 20 months,showed that the clinical symptoms in the two groups disappeared and the hydronephrosis relieved. Conclusion Compared with RLADP,the TRADP has the comparable operative time,but the postoperative management for TRADP is more simple and the healing is faster,the postoperative outcomes are comparable as well.
6.Effects of echinomycin on endothelin-2 expression and ovulation in immature rats primed with gonadotropins.
Zhengchao WANG ; Zhenghong ZHANG ; Yanqing WU ; Liyun CHEN ; Qianping LUO ; Jisen ZHANG ; Jiajie CHEN ; Zimiao LUO ; Xiaohong HUANG ; Yong CHENG
Experimental & Molecular Medicine 2012;44(10):615-621
Echinomycin is a small-molecule inhibitor of hypoxia-inducible factor-1 DNA-binding activity, which plays a crucial role in ovarian ovulation in mammalians. The present study was designed to test the hypothesis that hypoxia-inducible factor (HIF)-1alpha-mediated endothelin (ET)-2 expressions contributed to ovarian ovulation in response to human chorionic gonadotropin (hCG) during gonadotropin-induced superuvulation. By real-time RT-PCR analysis, ET-2 mRNA level was found to significantly decrease in the ovaries after echinomycin treatment, while HIF-1alpha mRNA and protein expression was not obviously changed. Further analysis also showed that these changes of ET-2 mRNA were consistent with HIF-1 activity in the ovaires, which is similar with HIF-1alpha and ET-2 expression in the granulosa cells with gonadotropin and echinomycin treatments. The results of HIF-1alpha and ET-2 expression in the granulosa cells transfected with cis-element oligodeoxynucleotide (dsODN) under gonadotropin treatment further indicated HIF-1alpha directly mediated the transcriptional activation of ET-2 during gonadotropin-induced superuvulation. Taken together, these results demonstrated that HIF-1alpha-mediated ET-2 transcriptional activation is one of the important mechanisms regulating gonadotropin-induced mammalian ovulatory precess in vivo.
Animals
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Cells, Cultured
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Chorionic Gonadotropin/*pharmacology
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Echinomycin/*pharmacology
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Endothelin-2/genetics/*metabolism
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Female
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Gonadotropins, Equine/*pharmacology
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Granulosa Cells/drug effects/metabolism
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Humans
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Hypoxia-Inducible Factor 1, alpha Subunit/*antagonists & inhibitors/genetics/metabolism/physiology
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Oligonucleotides/genetics
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Ovary/cytology/drug effects/physiology
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Rats
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Rats, Sprague-Dawley
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Superovulation/*drug effects
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Transcriptional Activation
7.Epidemiological characteristics and pathogenic analysis of herpetic angina in eastern region of Chengdu in 2022
Xiaoling TAO ; Xueyu WANG ; Min CHEN ; Xuchuan JIANG ; Zhengchao ZHANG ; Min HUANG ; Yu FU ; Xiaoyan FU
Chinese Journal of Experimental and Clinical Virology 2023;37(6):613-617
Objective:To study the epidemiological characteristics and pathogen status of infection of patients with herpetic pharyngitis in eastern region of Chengdu in 2022, in order to provide experimental basis for scientific prevention and control of herpetic pharyngitis.Methods:Information and fecal specimens were collected from patients diagnosed with herpetic angina at sentinel monitoring hospitals in eastern region of Chengdu from January to December 2022, their epidemiological characteristics were analyzed, real-time fluorescence quantitative PCR (RT-PCR) and semi nested RT-PCR (RT-snPCR) were used to detect and identify enterovirus subtypes.Results:Among the 236 cases of herpetic angina, the age of the affected children was mainly 0-9 years (99.15%), with children aged below 3 years accounting for 75.85% of the total cases, children in the 1-year-old group had the highest constituent ratio, accounting for 47.88% of the total cases; The virus detection rate in the 2-year-old group was higher than that in the 0-year-old group ( χ2=5.945, P=0.015). There were 142 male infected cases (60.17%) and 94 female cases (39.83%); there was a statistically significant difference in the virus detection rate between the male group (91.55%) and the female group (78.72%) ( χ2=7.938, P=0.005). In the seasonal distribution, the summer group had the highest constituent ratio (40.25%), while the winter group had the lowest constituent ratio (11.86%), The composition ratio of the summer group was significantly higher than that of the other groups, and the differences were statistically significant ( χ2=10.393, 19.199, 49.358, all P<0.001). The virus detection rate in the summer group was significantly higher than that in the spring group and the winter group ( χ2=16.951, 4.592, both P<0.05). The RT-PCR result showed that a total of 204 out of 236 children with herpetic angina were virus positive, with a detection rate of 86.44% (204/236), including coxsackievirus A (CV-A) 6 (22.55%), CV-A16 (5.88%), CV-A2 (2.94%), enterovirus type A71 (EV-A71) (2.94%), and other enteroviruses (65.69%). The RT-snPCR result showed that five viruses were identified from other enteroviruses, including CV-A4 (8.33%), CV-A5 (6.86%), CV-A10 (12.25%), CV-A12 (24.02%), and coxsackievirus B3 (CV-B3) (6.86%); another 15 positive samples of enterovirus were not successfully classified. We selected 9 strains of the virus in this study and 30 reference sequences from the NCBI database to. Conclusions:Herpetic angina in eastern region of Chengdu in 2022 is mainly found in male children under 3 years of age, and was concentrated between April and June. It has been confirmed that CV-A12 and CV-A6 were more common in intestinal viruses, and the 9 strains isolated in this study were homologous to multiple isolates.
8.Contribution of Wnt Signaling Pathway to Embryo Implantation
Yedong TANG ; Zhenghong ZHANG ; Zonghao TANG ; Zhengchao WANG
Acta Academiae Medicinae Sinicae 2018;40(1):91-97
The establishment of the endometrial receptive state,the implantation of activated blastocysts,and the decidualization of the uterine stroma are essential conditions for successful implantation in mammals and play decisive roles for subsequent pregnancy outcomes.The Wnt signaling pathway is associated with a variety of key functions such as proliferation,differentiation,apoptosis,and regeneration.During the embryo implantation,the Wnt signaling pathway plays an indispensable role.Studies have shown that the abnormal expression of Wnt signaling pathway can lead to endometriosis,endometrial cancer,and other reproductive diseases that seriously affect human health and quality of life.In this article we summarize the roles of Wnt signaling pathway in the establishment of uterine receptivity,blastocyst activation,embryo implantation,and decidualization and its regulatory mechanisms.
9.Clinical observation on 5 cases of small intestinal bezoars with obstruction treated by oral double-balloon enteroscopy
Liangliang SHI ; Zhengchao LI ; Shuling HUANG ; Jing GE ; Yuqin TONG ; Yiyang ZHANG
Chinese Journal of Digestive Endoscopy 2024;41(6):484-486
To evaluate the safety and efficacy of oral double-balloon enteroscopy for small intestinal bezoars with obstruction, the clinical, endoscopic, imaging, pathological and surgical data of patients with small intestinal bezoars and obstruction treated in Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from December 2021 to March 2023 were retrospectively analyzed. A total of 5 patients, all female, aged 52-83 years, were included. The clinical manifestation was mainly digestive tract obstruction. The diameter of bezoars was 30-50 mm. Three were located in the jejunum, one in the proximal ileum, and one at the jejuno-ileal junction. Three cases were diagnosed as having small intestinal bezoars by imaging examination before operation. All 5 patients achieved technical success. Four patients achieved clinical success, the symptoms of obstruction were completely relieved, and 1 patient was partially relieved due to malignant obstruction. No obvious complication occurred in 5 patients after operation. The follow-up time was 15-790 days, and there was no recurrence. The results showed that oral double-balloon enteroscopy for small intestinal bezoars was safe and effective, providing a new treatment for patients with small intestinal bezoars except for surgery.
10.Impact of unidentified injectable fillers on nasal dorsal soft tissue and rhinoplasty
Zhengchao LI ; Chen ZHANG ; Weiyan CHEN ; Bo HU ; Jieqing WANG ; Xiaowei SU
Chinese Journal of Medical Aesthetics and Cosmetology 2024;30(3):256-261
Objective:To explore the impact of unidentified injectable fillers on the soft tissue of nasal dorsum and rhinoplasty.Methods:The Plastic Surgery Information System of Xinhua Hospital Affiliated with Dalian University was utilized to conduct an analysis of 62 rhinoplasty patients between 2018 and 2019. Specifically, this included 28 patients with an unidentified history of injectable filler rhinoplasty, encompassing 1 male and 27 females with ages ranging from 19 to 53 years and a mean age of 28.8 years. Additionally, 34 patients underwent primary rhinoplasty, all of whom were female with ages ranging from 19 to 46 years and a mean age of 26.8 years. This study examined the effects of unidentified injectable fillers on the soft tissue of the nasal dorsum by analyzing the excised nasal dorsum under a microscope. Subsequently, statistical methods were performed to assess differences in gender, age, preoperative tip protrusion/nose length, postoperative tip protrusion/nose length, dorsal augmentation modality, and satisfaction, and to investigate the effect of unidentified injectables on the rate of dissatisfaction after rhinoplasty.Results:The histopathological analysis of unidentified injectable fillers removed from the nasal dorsum revealed the presence of mainly gel and granular fillers. The gel fillers, characterized by its pink jelly-like texture, contained unidentified injectable fillers, colorless glue, and were observed to flow out upon cutting. The granular filler, on the other hand, appeared as tough, irregularly shaped tissue similar to caviar. Additionally, evidence of muscle tissue in 5 pathologic sections indicated that the unidentified injectable fillers were injected into or near the dorsal nasal muscles, leading to varying degrees of muscle injuries upon excision. A comparison of 28 rhinoplasty patients with unidentified injectable fillers for nasal dorsal augmentation and 34 patients with primary rhinoplasty showed that 11 females in the former group and 4 females in the latter group were dissatisfied with the results. Statistical analysis demonstrated no significant differences between the two groups in terms of gender ( P=0.452), age ( P=0.219), preoperative tip projection/nasal length ( P=0.681), postoperative tip projection/nasal length ( P=0.105), and nasal dorsum augmentation methods ( P=0.413). However, the initial rhinoplasty group had a lower dissatisfactory rate (4 cases, 11.76%) and the unidentified injectables group had a higher dissatisfactory rate (11 cases, 39.29%), which was statistically significant between the two groups (χ 2=6.341, P=0.012). Conclusions:The presence of unidentified nasal injectable fillers can significantly decrease postoperative satisfactory rates, increase the incidence of dissatisfaction, and have adverse effects on the soft tissues of the nasal dorsum and the overall outcome of the rhinoplasty procedure.