2.Survival and melanogenic potential of reversibly immortalized human melanocytes mediated by SV40T antigen gene and Cre/loxP system in Guinea pigs
Ying WANG ; Zhihua ZENG ; Xichuan YANG ; Fei HAO ; Baiyu ZHONG
Chinese Journal of Dermatology 2010;43(3):188-191
Objective To study the survival and melanogenic potential of human melanocytes reversibly immortalized via SV40T antigen gene and Cre/loxP system in Guinea pigs. Methods The supernatants of retrovirus vector Cre-ERT2 were used to infect melanocytes which had been successfully transfected by SV40TAg gene (MCT), then the expression of Cre recombinase was induced with tamoxifen in infected cells; subsequently, the surviving cells, which were named as MCTC, were subjected to expansion culture. Guinea pigs were utilized to establish animal models of vitiligo, then MCTC and primary melanocytes were transplanted respectively into the animal models. The repigmentation at the transplanted area was observed with naked eyes successively until 3 months after the transplantation when tissue samples were obtained from implanted area and nonimplanted area of guinea pigs and subjected to Masson-Fontana silver stain and Hematoxylin-eosin stain for the analysis of melanocyte distribution and melanin deposition in epidermis. Results Repigmentation started 4 weeks after the transplantation, and dark or brown patches, which ranged in size from 0.5 to 1 cm, were observed in the implanted area 3 months after the transplantation. The repigmentation rate was of no significant difference between pigs transplanted with MCTC and those with primary melanocytes (82.5% vs 76.7%, P > 0.05). Pathological examination revealed melanin deposition in the basal layer of epidermis and some hair follicles in transplanted area. Conclusions SV40T antigen gene combined with Cre/loxP site-specific recombinase system can induce the reversible immortalization of human melanocytes, and the immortalized melanocytes have a favorable profile of biological safety and similarity in survival rate and melanogenic potential to primary melanocytes.
4.The study of influence on cyclin and cyclin-dependent kinase inhibitor of Kasumi-1 cell treated with VPA
Lei ZHAO ; Zhihua ZHANG ; Cuimin ZHU ; Wenliang TIAN ; Changlai HAO
Journal of Leukemia & Lymphoma 2009;18(1):5-8
Objective To explore the influence on cyclin and cyclin-dependent kinase inhibitor of cell cycle of Kasumi-1 cell-derived from t(8;21)/AML treated with histone deacetylase(HDAC)inhibitor valproic acid(VPA).Methods RT-PCR assay was applied to detect mRNA expression of regulation factor of cell cycle of Kasumi-1 cell with VPA at different concentration and different time. Results VPA could down-regulate mRNA expression of cyclinD1,cyclinE1 and cyclinB1,and up-regulate mRNA expression of p21 WAF1/CIP1 with no obvious variation of mRNA level of p27KIP1. Condusion VPA could regulate cell cycle through the regulation of cyclins and cyclin-dependent kinase,arresting Kasumi-1 cell at G0/G1 phase.
5.Application of pancreatic duct guide wire and transpancreatic septotomy with precutting techniques in dififcult endoscopic retrograde cholangiopancreatography
Shangbo JIN ; Yimin LIU ; Jidong HE ; Zhihua GUO ; Hao SUN
China Journal of Endoscopy 2016;22(12):75-78
Objective To investigate the application value of pancreatic duct guide wire and transpancreatic septotomy with precutting technique in difficult endoscopic retrograde cholangiopancreatography.Method258 patients who underwent difficult endoscopic retrograde cholangiopancreatography from April 2014 to April 2016 were selected as study subject, 128 patients among them received the technique of pancreatic duct guide wire, the other 130 patients received transpancreatic septotomy with precutting techniques. The success rate, intubation time and incidence of complications were compared between these two methods for cannulation.Result There was no signiifcant difference in preoperative clinical data between the two groups, the success rate did not differ signiifcantly between the two groups (93.75 % vs 93.85 %). Compared with transpancreatic septotomy with precutting techniques group, pancreatic duct guide wire group is less intubation time consuming (5.92 ± 0.69 vs 12.81 ± 3.67) min, the difference was statistically significant (t = -2.27,P < 0.05). 25 patients experienced complications, with 6 cases of acute pancreatitis, 2 cases of biliray tract infection in pancreatic duct guide wire group, and 8 cases of acute pancreatitis, 3 cases of hemorrhage,6 cases of biliray tract infection in transpancreatic septotomy with precutting techniques group. The pancreatic duct guide wire group had a signiifcantly lower incidence of complications (6.25 % vs 13.08 %). the difference was statistically signiifcant (χ2 = 3.27,P < 0.05). The incidence of acute pancreatitis did not differ signiifcantly between the two groups (4.69 % vs 6.15 %).ConclusionsPancreatic duct guide wire and transpancreatic septotomy with precutting techniques both can further improve the success rate of bile duct cannulation with ERCP. The incidence of acute pancreatitis did not differ significantly between two groups. But pancreatic duct guide wire group is less intubation time consuming, and had a significantly lower incidence of complications. Because of the convenience and safety of the pancreatic duct guide wire technique, and the insertion of the pancreatic duct does not increase the risk of postoperative acute pancreatitis. We think that this method is more worthy of Clinical promotion.
6.Clinical Effect Observation of Alendronate Combined with Jintiange Capsules in the Treatment of Postm-enopausal Osteoporosis
De CAI ; Zhixiu LIN ; Zhihua HUANG ; Hao LIN ; Qun LUO
China Pharmacist 2015;(8):1352-1354
To explore the clinical effect of alendronate combined with Jintiange capsules in the treatment of postmeno-pausal osteoporosis ( PMOP) . Methods:Ninety-eight cases of PMOP patients were randomly divided into three groups. The combina-tion treatment group (34 cases) was treated with alendronate tablets combined with Jintiange capsules, while the control group A (32 cases) and the control group B (32 cases) was respectively treated with alendronate tablets and Jintiange capsules, and the treatment course was six months. The changes of the clinical symptom score, bone mineral density ( BMD) , serum calcium, serum phosphorus, AKP, hepatorenal function and so on before and after the treatment were recorded. Results:After the six-month treatment, the clinical symptom score of the treatment group was much better than that in the two control groups(P<0. 05). BMD in the three groups was in-creased after the treatment(P<0. 05), and the increase in the treatment group was more notable than that in the two control groups(P<0. 05). Conclusion:Alendronate combined with Jintiange capsules can effectively improve BMD and relieve clinical symptoms in the patients with PMOP, which is worthy of promoted application.
7.Diagnosis and treatment of infantile congenital subglottic hemangioma
Lan CHENG ; Qi HUANG ; Hao WU ; Jun YANG ; Li CHEN ; Zhihua ZHANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;(15):693-696
Objective: To discuss the diagnosis and treatment of infantile congenital subglottic hemangioma. Method:Nine patients with recurrent stridor,feeding difficulties and laryngeal obstruction were diagnosed as con-genital subglottic hemangioma with bronchia endoscopy. The lesions were unilateral in 7 cases and bilateral in 2 ca-ses. Eight cases were treated by intralesional Pingyangmycin injection after tracheotomy. Patients were discharged with metal trachea and were followed up endoscopically 2 weeks thereafter. A second treatment would be required if the tumor reduced in size but not disappeared completely. Plugging tubes and extubation would be done when the tumor disappeared completely. One case with unilateral lesion underwent power-assisted tumor resection with-out tracheotomy. Result: Hemangioma vanished completely in 6 patients with unilateral subglottic hemangioma who were treated with single stage procedure. Tumor size was found reduced significantly after one procedure and van-ished completely after a second procedure in 2 cases with bilateral subglottic hemangioma. All of the 8 cases were extubated successfully. The patient recovered thoroughly in two weeks after power-assisted tumor resection. Con-clusion: Intralesional Pingyangmycin injection after tracheotomy can relief the airway obstruction quickly, shorten the nature course of hemangioma, shorten the period of wearing trachea, has no complication such as subglottic stenosis and is thought to be a safe and effective therapy for large size hemangioma. Power-assisted tumor resec-tion under good total anesthesia without tracheotomy combined with well hemostasis is effective, less injure, and suitable for small size hemangioma.
8.Ultrastructural change of chicken basilar papilla during a prolonged administration of kanamycin
Mingliang XIANG ; Hao WU ; Yun LI ; Zhihua ZHANG ; Qi HUANG ; Li CHEN
Chinese Journal of Tissue Engineering Research 2006;10(26):177-180,封三
BACKGROUND: Although some studies have been done on the functional change of cochlea in chickens during prolonged administration of kanamycin,few studies on ultrastructural changes in chicken cochlea have been reported.OBJECTIVE: To observe ultrastructural changes of the chicken basilar papilla following poisoning with prolonged administration of kanamycin.DESIGN: A completely randomized design and controlled observation.SETTING: Affiliated Xinhua Hospital, Shanghai Second Medical University.MATERIALS:The experiment was conducted in Xinhua Hospital Affiliated to Medical College, Shanghai Jiao Tong University from June 2001 to August 2003. Newly born Roman chickens (n=78), of either sex, provided by Shanghai Guixing Breeding Chick Farm, were used as subjects.METHODS: 200 mg/kg kanamycin was injected into muscles daily at day 3 after birth for 10 days. Then they were divided into 3 groups randomly:①typical application group: The chickens were applied with drugs no longer, and then killed at days 1, 3,7, 10 and 15, 6 chickens in each time phase. ②prolonged application group: 200 mg/kg kanamycin per day were injected into muscles continuously, and then killed at days 13, 17, 20, 25and 30, 6 chickens in each time phase. ③control group: The chickens were not treated with any drug. They were assigned into 3, 13 and 33 days groups with 6 chickens in each time phase. When the chickens were killed they were at the same age with the experime ntal animals at days 0, 10 and 30 of kanamycin treatment. All the experimental animals were studied with auditory brain-stem response (ABR) and scanning electron microscopy in chicken basilar papilla to observe ultrastructural change of cochlea, threshold, latency and amplitude of ABR wave I.MAIN OUTCOME MEASURES: ①Ultrastructural change of the chicken basilar papilla, and ② ABR result of chicken.RESULTS: A total of 78 Roman chickens were involved in the result analysis, without dropout. ①Outcome of ABR: Loss and recovery of chicken ABR threshold value was similar between within continuous administration for 20 days and administration for 10 days plus 10 days of recovery (drug withdrawal group). Twenty days after administration, ABR threshold significantly increased again. ②Outcome of scanning electron microscopy: At the end of 10-day kanamycin administration, hair cells in 40% proximal chicken basilar papilla were destroyedand disappeared completely. Although continuous administration was performed, the hair cells began to regenerate and repair after damage. Within the 20-day continuous administration, the regenerated hair cells of basilar papilla developed maturity, which was mostly coincident with the administration for 10 days group. At day 25, regenerated hair cells of proximal basilar papilla were injured once again. At day 30, most of the regenerated hair cells were destroyed and disappeared.CONCLUSION: Prolonged administration of kanamycin obviously prevents the full repair of chicken basilar papilla damaged by kanamycin poisoning.
9.Preventive effect of alprostadil on contrast-induced nephropathy in high risk patients after PCI
Hao YU ; Wei ZHU ; Yuhe KE ; Zhihua YU ; Liqun TIAN ; Gangfeng DUAN ; Fei WEN ; Qiongli ZHENG
Chinese Journal of cardiovascular Rehabilitation Medicine 2017;26(4):402-405
Objective:To observe influence of alprostadil injection on contrast-induced nephropathy (CIN) in high risk patients after percutaneous coronary intervention (PCI).Methods: A total of 263 CIN high risk (CIN risk score ≥16 scores) patients were selected.According to random number table, patients were randomly divided into routine treatment group (n=121, received routine hydration therapy) and alprostadil group (n=142, received additional alprostadil injection based on routine treatment group).Serum creatinine (SCr), glomerular filtration rate(GFR), cystatin C (CysC) and β trace protein (β-TP) level before, 48h and 72h after PCI were measured and compared, and incidence rate of CIN, percentage of blood purification therapy and mortality were compared between two groups.Results: Compared with before PCI, there was significant rise in SCr level and significant reduction in GFR in both groups on 48h and 72h after PCI (P<0.01 all);Compared with routine treatment group, there were significant reductions in levels of SCr [72h: (190.04±28.92) μmol/L vs.(141.10±21.18) μmol/L], and significant rise in GFR [72h: (26.0±4.4) ml/min vs.(36.4±4.9) ml/min], and levels of CysC[72h: (1.75±0.74) mg/L vs.(1.47±0.55) mg/L] and β-TP [72h: (1.53±0.50) mg/L vs.(1.22±0.38) mg/L] significantly decreased in alprostadil group on 48h and 72h after PCI, P<0.05 or <0.01;there were significant reductions in incidence rate of CIN (30.6% vs.18.3%) and percentage of blood purification therapy (10.7% vs.3.5%) in alprostadil group, P=0.001, 0.045 respectively.There was no significant difference in mortality between two groups, P=0.728.Conclusion: Alprostadil injection can significantly improve kidney function, reduce incidence rate of CIN and percentage of blood purification therapy in CIN high risk patients after PCI, which is worth extending.
10.Effect of different time neoadjuvant hormornal therapy for locally advanced prostate cancer
Ruipeng HOU ; Jian LI ; Fengwei WANG ; Zhihua ZHANG ; Hao WEI ; Tao TANG ; Hua ZHANG
Cancer Research and Clinic 2011;23(10):684-686
Objective To investigate ideal solution of neoadjuvant hormomal therapy (NHT) for locally advanced prostate cancer.Methods 60 patients diagnosed with locally advanced (T3-4N0M0) prostate cancer were treated with NHT.They were randomly divided into 3 groups of 20 cases.A group:NHT 2 weeks,B group:NHT 3 months,C group:NHT 6 months.Results The median PSA of A,B and C group after NHT were 24.88 (6.62-55.86),0.20 (0.05-12.07) and 0.07 (0.01-2.01) ng/ml,respectively.There was statistically significance compared with those in untreatment ( all P =0.00).There was statistically significant (P =0.00)among groups.The prostate volume of A,B and C group were (49.50+14.19),(47.35±17.99) and (36.15±7.17)ml,respectively.There was statistically significance in the B and C group compared with that in untreatment (P =0.04,0.00).There was statistically significant between A and C group and between B and C group (P =0.00,0.01).The Qmax of A,B and C group were (8.75±2.15),(11.7±2.81) and (14.45±2.61) ml/s,respectively.There was statistically significance in the B and C group compared with untreatment (both P =0.00).There was statistically significance among groups (all P =0.00).Conclusion The NHT time should last at least 3 months in order to reduce PSA and prostate volume and to increase the Qmax.