1.Establishment of coculture model of blood-brain barrier in vitro for nanoparticle's transcytosis and toxicity evaluation
Wei LU ; Yuzhen TAN ; Xinguo JIANG
Acta Pharmaceutica Sinica 2006;41(4):296-304
Aim A method of coculture of brain capillary endothelial cells (BCECs) and astrocytes of rats was used to evaluate nanoparticle' s blood-brain barrier (BBB) transcytosis and toxicity at the endothelial tight junction. Methods A lipophilic fluorescent probe, 6-coumarin, was incorporated in poly(ethyleneglycol)-poly (lactide) nanoparticle using double emulsion/solvent evaporation method. BCECs and astrocytes were firstly isolated from brain of newborn rats and characterized by their morphology and immunocytochemistry staining, separately. Subsequently, a coculture model with BCECs on the top of micro-porous membrane of cell culture insert and astrocytes on the bottom side was established. The permeability of 14C-labeled sucrose and nanoparticle were determined, separately. Results The meanweight-based diameter of 6-coumarin loaded nanoparticles was ( 102.4 ± 6.8) nm, with zeta potential of ( - 16.81 ± 1.05) mV. BCECs were positive for factor Ⅷ staining and glial fibrillary acidic protein was tight junction between BCECs in the coculture model could be visualized by both scanning electron microscopy and transmission electron microscopy. The unchanged paracellular transport of sucrose proved vivo situation for examination of the permeability of nanoparticle and toxicity evaluation.
2.Preliminary clinical experience of laparoscopic repair of esophageal hiatal hernia
Lijie TAN ; Dayong GU ; Wei JIANG
Chinese Journal of Minimally Invasive Surgery 2001;0(06):-
Objective To summarize the clinical experience of laparoscopic repair of esophageal hiatal hernia.Methods A total of 15 cases of esophageal hiatal hernia underwent laparoscopic hernia repair and fundoplication from May 2004 to April 2005 in this division.There were 4 cases of type Ⅰ hernia(all of which presented severe gastroesophageal reflux disease),10 cases of type Ⅱ hernia,and 1 case of type Ⅲ.Surgical procedures included laparoscopic Nissen total fundoplication in 9 cases,Toupet partial fundoplicatin in 4 cases,and Dor partial fundoplication in 2.Symptoms of gastroesophageal reflux disease,including heart burn,dysphagia,regurgitation,chest pain,and belching,were evaluated by using the Visual Analogue Scales(VAS) at preoperative period,1 postoperative month,and 6 postoperative months,respectively.Results No conversion to open surgery was required in this study.The operative time ranged 100~187 min(mean,125 min).The postoperative hospital stay was 2~5 days(mean,2.8 days).All the patients were followed for 1~12 months(mean,8.5 months).No hernia recurrence was found.The VAS scores decreased from 5.0?3.9 preoperatively to 0.9?1.3 at 1 month postoperatively(t=3.823,P
3.Investigation on medical cost of 72 patients with panic disorder
Wei-Xing CHEN ; Liang LI ; Qin-Lian ZHAO ; Jiang TAN ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(12):-
Objective To investigate the medical cost of 72 patients with panic disorder and efficacy of paroxetine(SSRIs)in the treatment.Methods The course,medical cost,types and times of main examination of all 72 patients were reviewed and analyzed.The effect of paroxetine on the patients was studied in terms of anxiety and depression scale.Results The 72 patients had long course,high medical cost and received many unnecessary medical examinations.Paroxerine was effective on patients with panic disorder in light of anxiety and depression scale.Con- clusion Patients with panic disorder had long course and spent much unnecessany money before final diagnosis. Paroxetine was useful and safe in the treatment of panic disorder.
4.Clinical significance of early diagnostic value of urinary neutrophil gelatinase-associated lipocalin in acute kidney injury in sepsis patients
Xingkai XU ; Liandong ZHANG ; Meichun TAN ; Hao JIANG ; Wei SHI
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2017;24(3):267-269
Objective To estimate the value of urinary neutrophil gelatinase-associated lipocalin (NGAL) level for early diagnosis of acute kidney injury (AKI) in patients with sepsis.Methods One hundred and twenty-six sepsis patients admitted to intensive care unit (ICU) in Baoshan Branch Hospital of Shuguang Hospital Affiliated to Shanghai University ofTraditional Chinese Medicine from June 2014 to December 2015 were enrolled, and they were divided into two groups according to whether complication of AKI was present. The levels of urinary NGAL in the two groups of septic patients were evaluated immediately and at 12, 24 and 48 hours after the definite diagnosis, and the levels were compared between the two groups; the receiver operating characteristic curve (ROC curve) was performed and the value of urinary NGAL level in early diagnosis of sepsis AKI was evaluated.Results There were 60 septic cases complicated with AKI (AKI group), with the prolongation of time after definite diagnosis, the urinary NGAL (g/L) levels were gradually increased at 12, 24 and 48 hours, the levels were significantly higher than those at the corresponding time points in the group without AKI [non AKI group (66 cases), 12 hours: 178.2±32.8 vs. 53.8±10.4, 24 hours: 228.4±24.6 vs. 54.1±9.0, 48 hours: 186.1±43.6 vs. 52.5±9.4, allP < 0.05]. The area under ROC curve (AUC) of urinary NGAL level at 24 hours after definite diagnosis and 95% confidence interval (CI) were 0.863 (0.766-0.929) and 0.686 (0.466-0.696), respectively, when the cutoff value of urinary NGAL was 65.9μg/L, the sensitivity was 81.9% and specificity 76.1%; when the cutoff value of urinary NGAL was 57.9μg/L, the sensitivity was 70.2% and the specificity 57.2%.Conclusion Urinary NGAL level can be used as a reference marker for the early diagnosis of sepsis concomitant AKI.
5.Effect of chronic rhGH stimulation on STAT5 in normal mouse liver and its molecular mechanism
Wei TAN ; Xiaolan CHEN ; Chuqiong WANG ; Bo JIANG ; Jiman HE
Chongqing Medicine 2013;(26):3142-3144,3148
Objective Toinvestigatetheeffectofchronicrecombinantgrowthhormone(rhGH)stimulationonhepaticSTAT5in healthy young mouse and its molecular mechanism .Methods 32 normal young mice were divided into 4 groups :PBS ,PBS+single rhGH ,chronic rhGH ,chronic rhGH+ single rhGH groups .The mice in the treatment groups(chronic rhGH and chronic rhGH +single rhGH )received intraperitoneal injection of rhGH at 1 μg/g body weight once daily for three weeks ;the mice in the control groups(PBS and PBS+single rhGH) were injected with sterile phosphate buffered saline(PBS ,0 .1 mol/L) .At next day 8:00 after 16 h of injection of last rhGH and PBS ,at 30 min before killing mouse ,PBS 0 .1 mol/L was injected in the PBS group and the chro-nic rhGH group ,while the single dose of rhGH 1 μg/g was injected in the PBS+single rhGH and the chronic rhGH+single rhGH group .Results Compared with the control mice(PBS group and PBS+single rhGH group) ,the body mass in the rhGH stimulation mice (chronic rhGH group and the chronic rhGH + single rhGH group) was significantly increased ;compared with the PBS group ,the base p-STAT5 level within the hepatic GH signal pathway in the chronic rhGH was significantly decreased ;the combi-ning capacity of hepatic STAT5 and DNA detected by EMSA in the chronic rhGH group was significantly lower than that in the PBS group ;in the mice injected by single dose of rhGH 1 μg/g at 30 min before killing mouse ,the hepatic p-STAT5 level in the chronic rhGH+single rhGH was significantly decreased compared with the chronic rhGH + single rhGH ;compared with the con-trol group ,the CIS content in the chronic rhGH was obviously increased ,while the SOCS-2 content had no obvious difference .Con-clusion Chronic rhGH stimulation can inhibit the phosphorylation of hepatic STAT 5 in normal young mouse ,which mechanism could be related with the increase of CIS content .
6.Effects of Astragaloside IV on Myocardial Fibrosis and Connective Tissue Growth Factor Expression in Experimental Rats With Chronic Heart Failure
Hongqiang JIANG ; Jinguo ZHANG ; Hongyong TAN ; Xiqing WEI
Chinese Circulation Journal 2016;31(2):165-169
Objective: To observe the effects of astragaloside IV on myocardial fibrosis and connective tissue growth factor (CTGF) expression in experimental rats with chronic heart failure (CHF).
Methods: CHF model was established by abdominal aorta constriction (AAC) and the rats were divided into 5 groups:Sham operation group, the rats received normal saline 2 ml/day, n=10, CHF group, the rats received normal saline 2 ml/day, n=12;Astragaloside IV groups, CHF rats received astragaloside IV at (20, 40, 60) mg/kg respectively and n=12 in each group. All animals were treated for 4 weeks. Hemodynamic indexes were monitored, left ventricular mass index (LVMI) was calculated, morphologic changes of myocardial tissue was observed by HE staining, myocardial ifbrosis degree and collagen volume fraction (CVF) were measured by Masson staining. The mRNA and protein expressions of CTGF were detected by RT-PCR and immunohistochemistry, Western-blot analysis respectivety.
Results: Compared with CHF group, 3 Astragaloside IV groups had decreased LVMI and CVF, P<0.05-0.01;Astragaloside IV (40 and 60) mg/kg groups presented decreased LVEDP and LVSP, increased ±dp/dtmax, P<0.01 to P<0.05 and improved pathological morphology. Compared with Sham group, CHF group had increased average CTGF OD value (0.09 ± 0.03) vs (0.45 ± 0.04) and increased expression of myocardial CTGF (0.57 ± 0.06);compared with CHF group, the average CTGF OD values in Astragaloside IV (20, 40, 60) mg/kg groups were all decreased (0.39 ± 0.05), (0.30 ± 0.06), (0.24 ± 0.04) and decreased expressions of myocardial CTGF (0.44 ± 0.05), (0.35 ± 0.02), (0.28 ± 0.03) respectively, all P<0.01.
Conclusion: Astragaloside IV can inhibit myocardial ifbrosis and improve cardiac function in CHF rats, which might be via inhibiting the over expression of myocardial CTGF.
7.Establishment of human neuroblastoma SH-SY5Y cell line stably silencing beclin1.
Chuanjie WEI ; Shuang XIAO ; Lan JIANG ; Yan TAN ; Bo HUANG ; Dingxin LONG
Journal of Biomedical Engineering 2014;31(5):1085-1089
The pGenesil-1-Beclin1 eukaryotic expression vectors were constructed to establish an SH-SY5Y cell line stably expressing shRNA-Beclin1. The shRNA was connected to pGenesil-1 to construct the recombinant plasmid pGenesil-1-Beclin1, which was transformed into JM109 E. coli. Positive clones were identified by digestion with restriction endonuclease and DNA sequencing. SH-SY5Y cells were cultured by the conventional method. The pGenesil-1-Beclin1 and pGenesil-1 plasmids were transfected into SH-SY5Ycells, and the cells were screened by G418 until the stable G418-resistant monoclonal cells were acquired. Beclin1 mRNA and Beclin1 protein were detected by RT-PCR and Western blot analysis respectively. The results of restriction endonuclease analysis and DNA sequencing confirmed the correct construction of the eukaryotic expression vector pGenesil-1-Beclin1. Two SH-SY5Y transfected cell lines were successfully selected. Compared with the control group, RT-PCR and Western blot showed that the expression of Beclin1 mRNA and protein were down regulated 71.28% ± 1.45% (P < 0.05)and 75.50% ± 2.63% (P < 0.05), respectively. The results indicated that the eukaryotic expression vector pGenesil-1-Beclin1 was successfully constructed and the SH-SYSY cell lines with inhibited Beclin1 expression were established. It provides a useful cell model for studying the biological function of Beclin1.
Apoptosis Regulatory Proteins
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genetics
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metabolism
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Beclin-1
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Cell Line, Tumor
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Down-Regulation
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Escherichia coli
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Gene Silencing
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Humans
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Membrane Proteins
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genetics
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metabolism
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Neuroblastoma
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Plasmids
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RNA, Messenger
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RNA, Small Interfering
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Transfection
8.Diagnosis and surgical treatment of pulmonary sequestration in adults
Bin JIANG ; Tianyu SUN ; Lingmin ZHANG ; Bo DENG ; Wei GUO ; Ruwen WANG ; Qunyou TAN
Journal of Regional Anatomy and Operative Surgery 2016;25(8):574-576
Objective To explore the diagnosis and surgical treatment of pulmonary sequestration in adults. Methods Clinical data of 21 cases of pulmonary sequestration whose diagnosis was confirmed by surgical biopsy in our department from March 2009 to February 2016 were retrospectively analyzed. Divided the patients into the thoracotomy group (n=9) and the thoracoscope group (n=12) according to dif-ferent surgical methods, and compared the diagnosis and surgery of the two groups. Results Among the patients, 8 cases were diagnosed as pulmonary sequestration and the remaining 13 cases were misdiagnosed,with the misdiagnosis rate of 61. 9%. Intraoperative exploration dem-onstrated that the abnormal blood vessels were originated from thoracic aorta (n=14,66. 7%),abdominal aorta (n=4,19%),phrenic artery (n=3,14. 3%) and aortic arch (n=1,4. 8%), and there were 20 cases (95. 2%) of intralobar sequestration and 1 case (4. 8%) of ex-tralobar sequestration. Patients underwent thoracotomy and patients underwent video-assisted thoracoscopic surgery were of no significant differences in operative time (P=0. 104),blood loss (P=0. 209),chest tube duration (P=0. 511),drainage volume (P=0. 135) and postoperative hospital stay (P=0. 450). All the patients recovered well after surgery. Conclusion As pulmonary sequestration lacks specific clinical manifestations,missed diagnosis and misdiagnosis are very common in patients. Chest enhanced CT and CT angiography are effective diagnostic methods at present. Both thoracotomy and VATS can achieve good therapeutic effects.
9.Reconstruction by the second toe tibial toe pulp skin flap for soft tissue defects of the finger
Jianjun LU ; Haitao TAN ; Jianzhong JIANG ; Xiang LUO ; Pingou WEI ; Han LIN ; Guiquan DENG
Chinese Journal of Microsurgery 2010;33(4):278-280,后插三
Objective To discuss the surgery procedure and treatment effect of reconstruction of the soft tissue of the thumb/finger defects by the second toe tibial toe pulp skin flap. Methods Ten patients with the soft tissue of pulp of the thumb/finger defects were treat by the same side of the second toe tibial toe pulp skin flap, all the patients have the soft tissue defect of finger pulp with exposed phalanx. Crush them in 4 cases, the machine cut wound in 6 cases. A fixed 2 cases, delayed operation 3-7d after injury to repair in 8 patients. The side of skin flap varied from 2.0 cm × 2.2 cm to 2.0 cm × 3.5 cm. Results Ten fingers in 10 cases all survived. Necrosis in edge part of the shin graft occurred in 2 cases, which was healed through changing of dressing. All cases were followed form 4 months to 16 months. The blood-supply, texture and elasticity of transferred flaps and the shape of fingers pulp were excellent. Good function recovery of the fingers was achieved. Pain and temperature sence were regained. Two point discrimination of the finger pulp was 5-9 mm.Primary healing occurred in all cases. It did not disturb dressing shoes and walking. Conclusion It is a reliable approach for soft-tissue coverage of the thumb/finger using the second toe tibial toe pulp skin flap based on distal perforators of digital artery or ulnar artery. The advantages include simply procedures, reliable blood supply without sacrificing main aneries and possibilities of sensoly recovery.
10.Free flap transplantation in the reconstruction of large area defects in head and neck
Xiang LUO ; Haitao TAN ; Jianzhong JIANG ; Jianjun LU ; Pingou WEI ; Keqin YANG ; Tianjin QIN ; Han LIN
Chinese Journal of Microsurgery 2010;33(5):357-359,后插3
Objective To review clinical experience of free flap transferring in the reconstruction of head and neck defects. Methods Fifteen free anterolateral thigh flaps, 5 free deep inferior epigastric flaps and 3 free lateral arm flaps were used to reconstruct large area head and neck defects. Results All 23 free flaps were alive with good appearance. 1 case survived by performing embolectomy after suffering arterial crisis and delayed recovery of wound took place in 2 cases. Conclusion Free flap transplantation is an ideal choice for reconstructing large area cranio-facio-cervical defects.