1.Construction of JSRV-NM Pseudovirions by High Efficiency Packaging System of the Lentivirus
Shumin GONG ; Guangming LI ; Zhimin WU ; Lizhen DONG ; Bin CHENG ; Bin ZHANG ; Ze ZHU
Tianjin Medical Journal 2014;(8):749-751
Objective To overcome the fact that SRV-NM virus can only multiple and amplify through partially pu-rified jaagsiekte retrovirus inoculated intratracheally in sheep but it cannot be augmented using in vitro cell culture, we con-structed JSRV-NM pseudovirions based on high efficiency packing system of lentivirus. Methods Lentivirus of three high efficiency packing plasmids system pMD.G, pCMV-HIV 8.2 and pHIV-eGFP was developed, and JSRV-NM-env coated plasmid pCMVJSRV-NM was used to substitute VSV-G virus coated plasmid pMD.G then co-transfected into 293T cells to replicate, package and produce restructured JSRV-NM pseudovirions. Gene expression of pseudovirion was determined through WPRE using real time PCR; Virus infectivity was detected through inoculating JSRV-NM pseudovirions into 24 pore plates. Results We construct JSRV-NM pseudovirions successfully based on the lentivirus system. JSRV-NM pseudo-virions can also be concentrated to higher titer (108 TU/mL detected by real time PCR by ultracentrifugation without signifi-cant loss of activity. JSRV-NM and VSV-G pseudovirions infected on Hela cells (both MOI= 3) respectively and no obvi-ous difference were shown on their infection efficiency detected by real time PCR. Conclusion Based on lentivirus system, JSRV-NM pseudovirions can be multipled and amplified in 293T cell culture in vitro. JSRV-NM pseudovirions is stable without loss its infection activity and the requirements of biological laboratory safety II was also met. JSRV-NM pseudoviri-ons will provide a useful tool for further study of JSRV-NM-env infection across species or its induction of lung adenocarci-noma.
2.Strategy for the diagnosis and treatment of bilateral testicular tumor
Da-Xin GONG ; Zhen-Hua LI ; Ze-Liang LI ; Xia WANG ; Shao-Bo YANG ; Jian-Bin BI ; Gang LI ; Zhi-xi SUN ; Chui-ze KONG
Chinese Journal of Urology 2001;0(10):-
Objective To evaluate the clinical features and the strategy for the diagnosis and treat- ment of bilateral testicular tumor.Methods The clinical data (including the signs and symptoms,imaging studies,tumor markers,treatment modalities and histopatbologic diagnoses) of 10 cases of bilateral testicular tumor from January 1980 to December 2004 were reviewed.Their age ranged from 19 to 58 years(mean,34 years).Of the 10 cases,8 with metachronous and 2 with synchronous testicular tumors were identified.The clinical stages at the primary and secondary tumor diagnosis were:5 cases of stageⅠ,3 of stageⅡ;and 6 cases of stageⅠ,1 of stageⅡ,and 1 of stageⅢ,respectively,in 8 metachronous tumor patients.Two syn- chronous tumor patients were both identified as stageⅠdisease.Histological examination showed the primary tumor (seminoma) in 4 cases and the secondary contralateral tumor (seminoma) in 3.Results Two syn- chronous tumor patients underwent bilateral radical orchiectomy simultaneously,and 8 underwent orchiectomy successively.Retroperitoneal lymph node dissection was performed in 3 cases.Postoperatively,hypogonadism occurred in 10 patients,and 7 of them received androgen replacement therapy.Follow-up ranged from 9 month to 23 years with a mean of 10.5 years.Two patients died of the disease;2 had metastasis (1 of them was alive with metastasis);2 had recurrences and underwent local resection.Conclusions Metachronous bilateral testicular cancers are more common than synchronous bilateral testicular cancers.Seminoma was the most common histopathologic type.Testis-sparing surgery can be performed in selected cases.
3.Wnt1/β-catenin signaling up-regulates spinal VGLUT2 expression to maintain neuropathic pain in mice
Zhi-Ling ZHANG ; Gang YU ; Xiao-Nan LIANG ; Rui-Bin SU ; Ze-Hui GONG
Chinese Journal of Pharmacology and Toxicology 2018;32(4):340-340
OBJECTIVE The present study was aimed to investigate the role of Wnt/β-catenin sig-naling in spinal VGLUT2 regulation and neuropathic pain. METHODS To elucidate the association be-tween VGLUT2 and neuropathic pain,we determined the expression and distribution characteristics of VGLUT2 in mice subjected to spared nerve injury(SNI),and then observed the effects of two VGLUT2 targeting shRNAs on mechanical allodynia and glutamate release.The effects of Wnt/β-catenin signal-ing on VGLUT2 expression and pain behavior were investigated by using Wnt agonist,Wnt1,and Wnt/β-catenin pathway inhibitor XAV939 in SNI mice.RESULTS SNI surgery induced significant up-regula-tion of VGLUT2 on postoperative days 7,14,and 21.Double immunofluorescence labeling of VGLUT2 with NeuN,MAP2,Iba-1,or GFAP showed that VGLUT2 was mainly expressed in neurons in the dor-sal horn of the spinal cord after SNI(NeuN,MAP2).Intrathecal administration of VGLUT2 shRNAs be-fore or after SNI surgery significantly decreased mechanical allodynia and glutamate release. Mean-while,Wnt1/β-catenin signaling increased significantly after SNI surgery.Over-expression of β-catenin in PC12 cells increased VGLUT2 protein level,intrathecal administration of Wnt agonist or Wnt1 signifi-cantly increased VGLUT2 protein expression in spinal cord, while Wnt/β-catenin pathway inhibitor XAV939 decreased VGLUT2 expression in PC12 cells and spinal cord.Additionally,intrathecal admin-istration of XAV939 7 days after SNI significantly attenuated mechanical allodynia in mice, which was in accordance with down-regulation of VGLUT2 protein levels.VGLUT2 shRNAs significantly attenuat-ed Wnt agonist or Wnt1 induced mechanical allodynia. CONCLUSION Wnt1/β-catenin signaling path-way up-regu-lates the spinal VGLUT2 expression,and this regulation is involved in neuropathic pain behavior.
4.Establishment and application of a mouse model for drug-induced schizophrenia.
Hui YAN ; Shu-Ling LI ; Rui-Bin SU ; Ze-Hui GONG
Acta Pharmaceutica Sinica 2013;48(4):484-488
Schizophrenia, described as the worst disease affecting mankind, is a severe and disabling mental disorder. Schizophrenia is characterized by complicated symptoms and still lacks a diagnostic neuropathology, so developing schizophrenia animal models which have quantifiable measures tested in a similar fashion in both humans and animals will play a key role in new therapeutic approaches. According to the symptoms of cognitive impairment and emotional disorder, the N-methyl-d-aspartate (NMDA)-receptor antagonist MK-801 was applied to induce schizophrenia-like behavior in mice. Locomotor activity and prepulse inhibition (PPI) were selected as indices and the effect of clozapine was also investigated in this model. The results showed that compared with the normal group, MK-801-treated mice exhibited significantly increased locomotor activity and impaired PPI, and pre-exposure to clozapine could ameliorate the abnormality and make it back to normal level. These findings suggest that the model we established could be a useful tool for antipsychotic drug screening.
Animals
;
Antipsychotic Agents
;
pharmacology
;
Clozapine
;
pharmacology
;
Disease Models, Animal
;
Dizocilpine Maleate
;
Inhibition (Psychology)
;
Male
;
Mice
;
Motor Activity
;
drug effects
;
Receptors, N-Methyl-D-Aspartate
;
antagonists & inhibitors
;
Schizophrenia
;
chemically induced
;
physiopathology
5.Clinical efficacy of daily low-dose tadalafil on erectile dysfunction-no sexual life.
Yu-Xi ZHANG ; Jian-Bin BI ; Da-Xin GONG ; Chui-Ze KONG
National Journal of Andrology 2013;19(6):542-544
OBJECTIVEErectile dysfunction-no sexual life (ED-NS) is defined as the inability to have enough penile erection hardness and duration so as to have enough confidence in attempting sexual intercourse for more than six months. This study was to investigate the effect of daily low-dose tadalafil on ED-NS.
METHODSWe treated 35 ED-NS patients aged 17-35 (25.9 +/- 3.9) years with oral tadalafil at 5 mg qd for 3 months and followed them up for another 3 months after drug withdrawal. We obtained the scores of the patients on Self-estimation Index of Erectile Function-No Sexual Life (SIEF-NS) and compared them before and after medication and at 3 months after drug withdrawal.
RESULTSThe patients' SIEF-NS scores were 43.2 +/- 7.1 after medication and 42.1 +/- 7.4 at 3 months after drug withdrawal, both significantly higher than 21.2 +/- 5.9 before treatment (P < 0.05), though there was no significant difference between the former two scores (P > 0.05).
CONCLUSIONDaily medication of low-dose tadalafil can significantly improve the erectile function of the patients with ED-NS.
Adolescent ; Adult ; Carbolines ; administration & dosage ; therapeutic use ; Erectile Dysfunction ; drug therapy ; psychology ; Humans ; Male ; Sexual Behavior ; Tadalafil ; Treatment Outcome ; Young Adult
6.Intraoperative localization of acetabular phase parameters and their clinical significance
bin Ze LI ; wei Tian LIN ; mei Xiao GONG ; qian Qian YI ; jia Lei SU ; Bing LI
Journal of Regional Anatomy and Operative Surgery 2017;26(12):859-862
Objective To study the significance of the related parameters of the abduction angle ,anteversion angle and summit arc angle in the intraoperative localization of anterior column and anterior wall acetabular fractures .Methods The safety angle of abduction angle ,an-teversion angle,summit arc angle and anterior wall acetabular of 318 adult pelvic specimen(109 males and 109 females) were measured.The data was statistical analyzed.Results For the male pelvic specimen,the measured values of abduction angle of acetabulum was (43.5 ± 2.9)°, anteversion angle was (13.7 ±5.2)°,summit arc angle inside was(25.5 ±7.3)°,summit arc angle front was(37.8 ±7.2)°,sum-mit arc angle back(49.9 ±12.7)°.For female pelvic specimen,the measured values of abduction angles of acetabulum was (44.1 ±3.2)° and anteversion angle was (14.3 ±4.9)°,summit arc angle inside was(27.1 ±6.5)°,summit arc angle front was(36.3 ±9.2)°,summit arc angle back was(52.3 ±13.6)°.According to '2-7' sector method, the thickness of different point of anterior wall was measured .There were statistically significant differences in abduction angles ,anteversion angle and summit arc angle (P<0.05).And there was no statistically sig-nificant difference in the thickness of anterior column and anterior wall between the above three one angles (P>0.05),there was no signifi-cant difference in the acetabular abduction angle ,anteversion angle and summit arc angle between males and females .Conclusion The intra-operative localization of acetabulum abducted angle ,anteversion angle and summit arc angle in the acetabulum anterior column and anterior wall fractures can achieve satisfactory therapeutic effect .
7.Study on multiple aldosterone-producing adenomas.
Xiu-Yue YU ; Chui-Ze KONG ; Zhen-Hua LI ; Zhi-Xi SUN ; Ze-Liang LI ; Jian-Bin BI ; Da-Xin GONG
Chinese Journal of Surgery 2007;45(24):1701-1703
OBJECTIVETo investigate the experience on diagnosis and treatment of multiple adrenal aldosterone-producing adenomas (APA).
METHODSEighteen cases of multiple adrenal APA were analyzed retrospectively, which were admitted from October 1992 to April 2006.
RESULTSAdrenalectomy was performed for 4 cases of unilateral synchronous multiple APA, which were discovered with three adenomas by 3D-CT; bilateral tumor resection was performed for 6 cases of bilateral synchronous multiple APA. There were 8 cases of bilateral metachronous multiple APA, including 2 cases of ipsilateral recurrent adrenal APA after adrenal tumor removal, which underwent tumor resection. Another 6 cases were contralateral APA following adrenalectomy due to adrenal APA, and underwent tumor resection. After operation, the adrenal function seemed to be normal, and no recurrence had been found on follow-up.
CONCLUSIONSUnilateral multiple synchronous APA require adrenalectomy. Tumor resection should be performed for bilateral or asynchronous APA, and it is very important to preserve healthy adrenal tissue as much as possible. 3D-CT has much value on diagnosis of small APA, unilateral multiple synchronous APA and ipsilateral recurrent adrenal APA.
Adenoma ; complications ; diagnosis ; surgery ; Adrenal Gland Neoplasms ; complications ; diagnosis ; surgery ; Adrenalectomy ; Adult ; Aldosterone ; blood ; Female ; Follow-Up Studies ; Humans ; Hyperaldosteronism ; blood ; etiology ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed
8.Stable co-expression of human kappa opioid receptor and PKAcat-EGFP in CHO cells and functional identification
Yu-Lei LI ; Long LONG ; Quan WEN ; Li-Li WANG ; Ze-Hui GONG ; Rui-Bin SU
Chinese Pharmacological Bulletin 2018;34(9):1321-1326
Aim To establish a cell model which stably co-ex-press human kappa opioid receptor (hKOR) and enhanced green fluorescent protein ( EGFP) labeled catalytic domain of cAMP-dependent protein kinase A(PKAcat) fusion protein (PKAcat-EGFP) in Chinese hamster ovary(CHO) cells, laying the foun-dation for the high-throughput screening of hKOR drugs and drug molecular mechanisms in vitro. Methods Hygromycin B resist-ant hKOR recombinant plasmid [ pcDNA3.1/Hygro ( + ) -hKOR] was transfected into CHO cells stably expressing PKA-cat-EGFP by a lipofectin based method. Transfected cells were selected in culture medium containing hygromycin B. The posi-tive clones were selected by PKA redistribution assay. Z’ factor was used for evaluation and validation the reliability of the cell model. PKA redistribution assay and LANCE cAMP 384 Kit were used to test the function of the receptors in selected clone. Results CHO-PKAcat-EGFP/hKOR-13 cell model exhibited stable response in PKA redistribution assay and LANCE cAMP 384 Kit. Treated with 100 nmol·L-1U-50488 for 30 min, the average value of Z’ factor was 0.596, proving the reliability of the cell model. The hKOR expression in cell model remained stable after a few generations. Conclusion The CHO-PKAcat-EGFP/hKOR-13 cell model with stable co-expression of hKOR and PKAcat-EGFP has been successfully established.
9.Rivaroxaban with aspirin for the secondary prevention of cardiovascular events in Chinese patients with stable cardiovascular diseases: subgroup analysis of COMPASS.
Yan LIANG ; Ze Bin GONG ; Ke Jia LOU ; Li Sheng LIU ; Jun ZHU
Chinese Journal of Cardiology 2021;49(9):873-879
Objective: This analysis was performed to evaluate the efficacy and the safety of rivaroxaban-aspirin combination therapy in secondary prevention of major adverse cardiovascular events in Chinese patients enrolled in the COMPASS trial. Methods: COMPASS was a prospective, international multi-center and randomized controlled trial. From September 2014 to February 2017, 1 086 patients with stable coronary artery disease and peripheral artery diseases were recruited from 31 centers in China. Patients were randomly assigned to separately receive the therapy of rivaroxaban (2.5 mg twice a day) plus aspirin (100 mg once a day,) group (n=366), rivaroxaban (5 mg twice a day) alone group (n=365), and aspirin (100 mg once a day) alone group (n=355). Baseline information such as age, sex, etc. of all three groups was collected. Finally, 1 081 patients were followed up successfully, with the follow-up rate 99.5% and the average follow-up time was 19 months. The primary efficacy endpoint was the composite of cardiovascular death, myocardial infarction and stroke. The primary safety endpoint was major bleeding evaluated by modified International Society on Thrombosis and Haemostasis criteria. Results: Age of patients was (64.2±8.3) years and there were 293 male in rivaroxaban plus aspirin group. Age of patients was (63.8±9.0) years, and there were 301 male patients in rivaroxaban alone group. Age of patients was (63.6±8.8) years, and there were 282 male patients in the aspirin alone group. The incidences of primary efficacy endpoint occurred in 9 cases (1.5%) in rivaroxaban with aspirin group, 21 cases (3.7%) in rivaroxaban alone group and 14 cases (2.5%) in aspirin alone group. Meanwhile, the incidences of primary safety endpoint occurred in 6 cases (1.0%) in rivaroxaban with aspirin group, 9 cases (1.6%) in rivaroxaban alone group and 7 cases (1.2%) in aspirin alone group. The net clinical benefit events were 10 cases (1.7%) in rivaroxaban with aspirin group, 22 cases (3.9%) in rivaroxaban alone group and 15 cases (2.7%) in aspirin alone group (P>0.5%). Conclusions: The combination of rivaroxaban with aspirin can be safe and effectively used for the secondary prevention in Chinese patients with stable coronary artery disease and peripheral artery diseases.
Aged
;
Aspirin/therapeutic use*
;
Cardiovascular Diseases/prevention & control*
;
China
;
Drug Therapy, Combination
;
Factor Xa Inhibitors/therapeutic use*
;
Female
;
Humans
;
Male
;
Middle Aged
;
Platelet Aggregation Inhibitors/therapeutic use*
;
Prospective Studies
;
Rivaroxaban/therapeutic use*
;
Secondary Prevention