1.Mechanism of radiosensitizing effects of endostatin on H-520 human lung squamous cancer cells
Zhenyu YOU ; Yong ZHAO ; Ping JIANG ; Na MENG ; Junjie WANG
Chinese Journal of Radiological Medicine and Protection 2010;30(3):287-290
Objective To investigate the mechanism of radiosensitizing effects of endostatin on H-520 human lung squamous cancer cells.Methods H-520 cells was treated with endostatin and/or radiation.Colony-forming assays were used to indicate the radiosensitising effects.Cell cycle distribution and expression of phosphor-p38-MAPK were assayed by FCM,and cyclin D1,cdk2,cdk4 and survivin mRNA leveh were assayed by RT-PCR.Phosphor-Akt was evaluated by Western-blotting.Results Combination of endostatin and irradiation inhibited the proliferation of H-520 cells.According to the colony-forming assays,the D0,Dq,D10 and SF2 values of the combination groups were much lower than those of irradiation groups.The sensitization enhancement ratio(SER)was 1.51.G2/M arrest occurred after 4 Gy irradiation.The gene expression of cyclin D1,cdk2,ckd4 and survivin and phosphor-Akt protein were down-regulated after treatment.The expression of phosphor-p38-MAPK protein was also down-regulated after treatment with 200 μg/ml endostar.Conclusions Endostatin inhibits the growth of H-520 cells and radiosensitizes the cells by induction of G0/G1 arrest,cell apoptosis and down-regulation of gene expression of cyelin D1,cdk2,cdk4 and reduces the phosphorylation of Akt and p38-MAPK.
2.Hypoxic preconditioning inhibits apoptosis of bone marrow mesenchymal stem cells through overexpressing Pim-1
You ZHANG ; Weiya YAN ; Zhengya SHEN ; Junjie YANG ; Jie HUI
Chinese Journal of Tissue Engineering Research 2016;20(14):1989-1998
BACKGROUND:Bone marrow mesenchymal stem cel s have a low survival rate after implanted into the ischemic myocardium. However, hypoxia preconditioning (HPC) may enhance bone marrow mesenchymal stem cel proliferation and promote its survival rate. OBJECTIVE:To explore whether Pim-1 is involved in HPC protecting against apoptosis of bone marrow mesenchymal stem cel s and the relevant mechanism. METHODS:Bone marrow mesenchymal stem cel s were respectively subjected to HPC for 0, 6, 12, and 24 hours. The expression of Pim-1 and apoptosis-related genes were detected by RT-qPCR and western blot. Then, the best hypoxic preconditioning time was determined as 12 hours. Then, bone marrow mesenchymal stem cel s were assigned to one of the fol owing groups:control (without HPC), 12-hour HPC, 12-hour HPC+Pim-1 inhibitor groups. Flow cytometry analysis was used to detect the cel apoptosis, Transwel assay to analyze the cel migration ability in each group, and JC-1 kit to detect mitochondrial membrane potential. Animal models of myocardial infarction were established. One week after modeling, bone marrow mesenchymal stem cel s were given via multi-point injection around the infarct zone of rats. Two weeks after modeling, heart tissues of rats were taken and sliced fol owed by DiI staining to calculate the survival rate of bone marrow mesenchymal stem cel s. Additional y, rat cardiac function was assessed by echocardiography prior to and after modeling as wel as at 4 weeks after cel transplantation. RESULTS AND CONCLUSION:At 12 hours after HPC, the expression of Pim-1, p-Akt and Bcl-2 gene in the infarct region was significantly increased, but the expression of caspase-3 and Bax was significantly decreased. Compared with the control group, cel viability in the 12-hour HPC group was increased very significantly at 1 week after cel transplantation (P<0.001), the migration and anti-apoptosis ability were enhanced significantly (P<0.01) and the cardiac function of rats was significantly improved in the 12-hour HPC group (P<0.05). Al of these protective effects were blocked by the Pim-1 inhibitor. These findings indicate that HPC can protect bone marrow mesenchymal stem cel s from apoptosis through activating Akt and up-regulating Pim-1, and thereby improve the therapeutic effect of bone marrow mesenchymal stem cel transplantation on ischemic heart diseases.
3.TACE in patients with hepatocelluar carcinoma feeded by left inferior phrenic artery
Hongping WU ; Junjie WANG ; Yong LOU ; Kaizhi YOU
Chinese Journal of Interventional Imaging and Therapy 2009;6(4):327-329
Objective To evaluate the efficacy of transcatheter arterial chemoembolization through left inferior phrenic artery (LIPA) in hepatocellular carcinoma (HCC) feeded by LIPA. Methods Angiographic data of 187 HCC patients were retrospectively analyzed. Five patients with LIPA feeding HCC were diagnosed and successfully treated with TACE through LIPA. Results All 5 patients underwent CT and angiography after 3 months, 3 of them achieved complete remission. LIPA recanalized in the other 2 patients, then these 2 patients underwent TACE through LIPA again. Conclusion LIPA could formed lateral branch to feed HCC. TACE through LIPA is safe and effective.
4.Radiosensitization of recombinant human endostatin in human lung squamous cancer cells in vitro
Zhenyu YOU ; Junjie WANG ; Yong ZHAO ; Hongqing ZHUANG ; Feng LIU ; Yingdong ZHANG
Chinese Journal of Radiation Oncology 2009;18(4):326-329
Objective To investigate the radiosensitising effect of recombinant human endostatin (endostar) on human lung squamous cancer cell line H-520 in vitro and its mechanism. Methods H-520 cells in exponential growing phase were treated with endostar alone, irradiation alone, or endostar plus irra-diation. Colony-forming assay was used to investigate the cytotoxicity and radiosensitising effects of endostar. Cell survival fractions of all groups were calculated and cell survival curves were fitted by single-hit multi-tar-get model. Cell apoptosis, cell cycle distribution and activated Caspase expression level were investigated by flow cytometry. Results The D0, Dq, D10 and SF2 values of combined treatment group were much lower than those of irradiation alone group. The sensitization enhancement ratio (SER) was 1.50 (ratio of D0 values). Endnstar induced H-520 cell apoptosis in a dose dependant manner. After administration of endostar, H-520 cell proliferation was inhibited, and cell apoptosis rate and apoptotic bodies were increased. After irradiation of 0 Gy, 2 Gy, 4 Gy and 8 Gy, the apoptosis rate of H-520 cells was 4.27% ±0.29%, 14.3% ±1.15%, 28.49% ± 1.58% and 54.79% ± 1.89% in the radiotherapy alone group, and 22.38% ± 1.61%, 35.01% ±1.16%, 46.83%±2.06% and 64.08%±4.28% in the combined treatment group, respective-ly. The difference between the two groups was significant (t = 19.17, 17.79, 25.64 and 3.44,all P < 0.05 ). Flow cytometric analysis showed that cell cycle distribution changed and G0 + G1 phase arrest oc-curred after endostar treatment, while irradiation induced G2 + M arrest. The expression level of activated Caspase in combination group (62.7% ±1.9% ) was higher compared to the control group ( 12.1%± 0. 1% ) , endostar alone group ( 54.6% ±1.0% ) and irradiation alone group ( 34.1%±1.2% ) ( t = 46.69, 6.55 and 22.54 ; all P < 0.05 ). Conclusion Endostar can enhance the radiosensitivity of H-520 ceils by inhibiting cell proliferation, promoting cell apoptosis and facilitating cell cycle redistribution.
5.Radiosensitization of microRNA-17-92 on human mantle cell lymphoma cells in vitro
Ping JIANG ; Junjie WANG ; Yong ZHAO ; Enyu RAO ; Na MENG ; Zhenyou YOU
Chinese Journal of Radiological Medicine and Protection 2010;30(4):403-406
Objective To investigate the effects of microRNA-17-92 on radiosensitivity of human mantle cell lymphoma cells. Methods Tetracycline-regulated pRevTet-On expression system was established to generate cell line Z138c-miR-17-92 with over-expressed miR-17-92 and cell line Z138c-TMP2. Cell proliferation was measured by 3 H-TdR incorporation and viable cell counting stained with typan blue. Cell cycle distribution was analysed by flow cytometry(FCM). Results More viable and proliferous cells were counted in group miR-17-92,when exposure dose was greater than 2 Gy and incubation time was longer than 48 h under the same condition (t = -3. 12 and -3.28,P <0. 05). The percentage of G2/M cells in group TMP2 was increased while no obvious cell cycle arrests were found in group miR-17-92 at 2 and 4 Gy (t = 2. 885, P < 0.05 ). When cells were incubated for 96 h, higher percentage of propidium iodide (PI) positively stained cells were found in group TMP2 (24. 02% vs. 36. 16% )compared with group miR-17-92 (6.49% vs. 11.39% ) at 2 and 4 Gy, respectively( t = - 17.59, - 4. 972, P < 0.05 ).Conclusions Overexpression of microRNA-17-92 decreased the radiosensitivity of human mantle cell lymphoma cells by inhibition of cell cycle changes and cell apoptosis.
6.Implementation of hospital information system based on VMware virtualization
Jun ZHANG ; Huang YU ; Xiangfeng WANG ; Junjie LI ; Jian YOU ; Bo JIANG ; Zheng YANG
Chinese Journal of Hospital Administration 2013;(2):108-110
Contemporary hospital information systems normally contain multiple applications,which are traditionally deployed as one application per server.More servers in the system will incur high system costs,low server efficiency and poor management and maintenance.The information system reform project of Wuxi Maternal and Child Health Hospital called into play the virtualization based on VMware.The project brought forth a number of merits,namely sizably raising efficiency of servers integration,simplifying server group management,reducing cost and promoting hospital informationization.
7.Analysis of the Application of Antifungal Drugs in Haematologic Patients from a Third Grade Class A Hos-pital during 2013-2014
Weihua DONG ; Junjie TANG ; Xianting XIE ; Haisheng YOU ; Yan WANG ; Yalin DONG
China Pharmacy 2016;27(5):587-590
OBJECTIVE:To provide reference for the formulation of correct antifungal treatment strategy,and to promote stan-dard use of antifungal agent. METHODS:A retrospective survey was conducted for 138 haematologic patients from May 2013 to May 2014 in a third grade class A hospital,of whom all had used antifungal drugs during hospitalization. We collected all patients' information and analyzed it statistically. RESULTS:Of 138 haematologic patients,3 were proven IFD (all were Candida infec-tion),6 were probable IFD,12 were possible IFD,and 117 were undefined IFD. The positive rates of fungi pathogenic detection, fungal smear,G-test,and GM-test were 15.3%,9.4%,6.4% and 23.4% respectively. 6 kinds of antifungal were used,and vori-conazole had the highest frequency,followed by fluconazole,itraconazole,amphotericin B,caspofungin and micafungin. 62.3%patients used only one kind of antifungal,but 15.9% patients used 2 or more kinds of antifungal. The average medication course was 20.5 days(1 day to 125 days). Irrational drug use showed improper drug selection,unreasonable dose,and replacing antifun-gal with insufficient basis. CONCLUSIONS:The antifungal use in haematologic patients in the hospital is consistent with the re-quirements of guidelines,but there are still some issues as insufficient antifurgal drug treatment course to be further standardized.
8.Repeatability of OPD-Scan Ⅲ and its consistency with Pentacam in measuring keratometry and astigmatism
Panpan LI ; You YUAN ; Lili HUANG ; Yan ZHU ; Junjie LI ; Yu SONG
Chinese Journal of Experimental Ophthalmology 2021;39(3):238-242
Objective:To analyze the repeatability of keratometry and astigmatism values measured by the OPD-Scan Ⅲ and the agreement of the parameters measured by OPD-Scan Ⅲ and Pentacam.Methods:A diagnostic test study design was adopted.Fifty patients (100 eyes) with refractive errors, aged from 21 to 35 years old, were selected from Second Affiliated Hospital of Nantong University and Lixiang Eye Hospital of Soochow University during August 2018.Spherical equivalent, astigmatim degree and axis were measured by Autorefraction.Corneal biometric measurements were measured three times continuously with the above two instruments.Keratometry values at the flat axis (K1), keratometry values at the steep axis (K2), astigmatim degree, axis, vector parameters J0 (Jackson cross cylinder at 0°or 180°) and J45 (Jackson cross cylinder at 45°) were recorded.Intraclass correlation coefficient (ICC) was used for repeatability analysis.Wilcoxon signed rank test, Spearman correlation analysis and Bland-Altman graphs were employed to analyze the comparability.This study adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Lixiang Eye Hospital of Soochow University (No.SLER2018112). Written informed consent was obtained from each patient prior to any examination.Results:The ICC of K1, K2, astigmatism, astigmatic axis, J0 and J45 measured by OPD-Scan Ⅲ were all greater than 0.900; the ICC of the astigmatism measured by Pentacam was 0.896, and the ICC of the other parameters measured by Pentacam were greater than 0.900; The values of K2, astigmatism, J0 and J45 measured by OPD-Scan Ⅲ were greater than those measured by Pentacam, and the differences were statistically significant (all at P<0.05). The values of K1, K2, astigmatism degree, axis, J0 and J45 measured by OPD-Scan Ⅲ were positively correlated with those measured by Pentacam (r s=0.981, 0.982, 0.900, 0.737, 0.921, 0.703, all at P<0.01). The 95% agreement of limits (LOA) of K1, K2, astigmatism, axis, J0 and J45 measurement difference between OPD-Scan Ⅲ and Pentacam were -0.52-0.50 D, -0.39-0.59 D, -0.37-0.48 D, -17.29°-20.38°, -0.12-0.24 D and -0.22-0.28 D, respectively. Conclusions:OPD-Scan Ⅲ has high credibility in measuring corneal refractive power and astigmatism degree, but its 95% LOA of astigmatism axis is too large to be accepted clinically.
9.Expression of Human Brain Derived Neurotrophic Factor Gene in E. coli
Zhimin LIU ; Junjie CHEN ; LinJia ; Ruohan WANG ; Leran YOU ; Yunhua DONG
Journal of Biomedical Engineering 2001;18(1):68-71
The primers specific for the full-length BDNF coding sequence was designed and synthesized. The BDNF coding sequence was directly amplified from human genomic DNA by using PCR and inserted into vector pGEM-3Zf(+). The recombinant DNA was transformed into the host cells JM109 to obtain the positive clone pGEMBF18. The restriction enzyme analysis and DNA sequence detection confirmed that the inser ted fragment of clone pGEMBF18 is the full-length BDNF coding sequence. The hBD NF DNA fragment was recovered from the clone pGEMBF18 and ligated with prokaryot ic expression vector pGEX-5T to construct the recombinant expression plasmid p5 TBF34. The E.coli JM109 transformed with p5TBF34 was induced with IPTG. A new pr otein band with apparent molecular weight 43 kDa was detected in the lysate of t he transformed cell by using SDS-PAGE. The result of western hybridization show ed that this fusion protein reacted specifically to the antibodies to human BDNF . The amount of the soluble fusion protein was about 503.04mg/L lysate, 7.53% of total bacterial soluble protein of transformed cells, estimated by absorbance sc anning of SDS-PAGE and protein quantitation.
10.Treatment of diabetic foot with antibiotic bone cement combined with delayed lateral supramolleolar perforator fascial flap
Ping HUANG ; Xiaofang YU ; Jiarui YOU ; Jinmei HE ; Guojun DU ; Yi LIU ; Xueli LI ; Junjie QUAN
Chinese Journal of Microsurgery 2022;45(2):128-132
Objective:To explore the clinical effect of antibiotic bone cement combined with delayed lateral supramolleolar perforator fascial flap in the treatment of diabetic foot(DF).Methods:From April 2020 to July 2021, a total of 6 patients with DF were treated with antibiotic bone cement combined with delayed lateral supramolleolar perforator fascial flap. The patients were 5 males and 1 female, aged from 45 to 67 years old with an average of 56.2 years old. The wounds were all located in dorsal foot, 4 in right foot and 2 in the left. The wound area was 2.4 cm×5.0 cm-6.5 cm×10.0 cm. The depth of wound were: 3 cases up to tendon layer, and 3 cases up to metatarsal bone. Two of the wound were complicated with metatarsal osteomyelitis. The wounds at Wagner grade 3 in 4 patients and grade 4 in 2 patients. The flap size was 3.0 cm×6.0 cm-8.0 cm×11.0 cm. All of the wounds were repaired with delayed supramolleolar perforator fascia flap after debridement, application of antibiotic bone cement and fumigation with Sanhuang decoction(a traditional Chinese medicine). The affected limbs were externally fixed with plaster and raised after surgery, and the colour, temperature, tension and capillary reaction of the flaps were closely observed. Stitches were removed 2 weeks after surgery and rehabilitation of the affected limb was performed. Regular follow-up was made postoperatively. The appearance of flaps and the scar of donor and recipient sites were observed. The foot and ankle function were evaluated by the American Orthopaedic Association foot and Ankle Surgery(AOFAS) score scale.Results:Six cases of DF had no recurrence of wound infection. All flaps survived well. The average follow-up time was 6(3-14) months. The postoperative follow-up revealed satisfactory appearance of the flap, only linear scars remained in the donor and recipient sites. The function of foot and ankle recovered well with full weight-bearing and normal walk. AOFAS scores ranged from 81 to 95.Conclusion:It is an effective method to treat DF by applying antibiotic bone cement combined with delayed superior lateral malleolus perforator fascial flap. The operation is simple, safe and can cut down the time of treatment, quickly control the wound infection. It deserves further trials.