1.Application of Antibacterial Drugs for Special Use and Bacterial Resistance in Our Hospital before and af-ter the Implementation of Network Online Approval Process
China Pharmacist 2017;20(3):509-512
Objective:To analyze the indicators and resistance rate of antibacterial drugs for special use in our hospital before and after the implementation of network online approval process to provide reference for the management and rational use of antibacterial drugs for special use. Methods:A retrospective analysis was performed, and the consumption amount, sales amount, utilization ratio, AUD, submission rate of microbial specimen and resistance rates for the main pathogenic bacteria of antibacterial drugs and ones for special use were investigated during 2012 and 2015 in our hospital. Results:The approval process of antibacterial drugs for special use developed from handwork to network online. The consumption amount of antibacterial drugs increased slightly year by year from 2012 to 2015 in our hospital, the mean expense of antibacterial drugs for the inpatients increased from 1602. 85 yuan to 1888. 63 yuan,and AUD increased from 54. 50 DDDs/(100 persons × d) to 65. 47 DDDs/(100 persons × d). The sales amount proportion of antibacterial drugs for special use (13. 90%) was the lowest in 2013, and the highest (17. 34%) in 2015, and AUD increased from 4. 85 DDDs/(100 person × d)to 6. 37 DDDs/(100 person × d), and the submission rate of microbial specimen before the treatment increased from 85. 5% to 90. 0%. The main pathogenic bacteria were Staphylococcus aureus, Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa and Acinetobacter baumannii,and their ratio had slight change. Except Staphylococcus aureus, the resistance rate of the other four pathogenic bacteria kept increasing, and the resistance rates of Acinetobacter baumannii to cefepime and carbapenems were over 50%. Conclusion:The approval process management of antibacterial drugs for special use must be strengthened in order to truly a-chieve reasonable use of antibacterial drugs for special use.
2.CHARACTERIZATION,ISOLATION AND CULTURE OF OSTEOCLASTS FROM NEONATAL RAT LONG BONES
Weizhen ZHANG ; Shifeng YU ; Linfan ZHENG
Acta Anatomica Sinica 1955;0(03):-
Osteoclasts were isolated mechanically from neonatal rat long bones and cultured on glass or devitalized human cortical bone slices. Both morphological and functional studies showed that the isolated cells were of typical osteoclasts, because; 1. they attached rapidly on the matrix and had pseudopodial activity when cultured on glass;2. they had multinuclei,most of them contained 3-10 nuclei;3. calci-tonin and clacitonin gene related peptide directly inhibited the cellular activity; 4. they showed high acid phosphatase activity; 5. resorption lacunae could be found when the cells cocultured with devitalized bone slices.
3.Construction of C3a secrectory expression vector and tubular epithelial cell line stably expressing C3a
Jingmin ZHENG ; Guang YIN ; Shifeng YUN ; Wenjin ZHAO
Journal of Medical Postgraduates 2015;(6):584-589
[Abstract ] Objective Evidence from previous studies indicated that over-activation of C3a/C3aR axis existed in the renal tubular epithelial cells of patients with renal diseases including diabetic nephropathy .However , the pathological significance of over-ac-tivating C3a/C3aR axis still remains to be elucidated .In this study, we constructed a renal tubular epithelial cell line over-expressing C3a in a secretory manner in order to provide a cell model to investigate the pathological significance of over -activating C3a/C3aR axis under various pathological scenes . Methods We designed a synthesized C3a secretory expression unit and cloned it into the multi-clonal site of lentivirus expression vector pLenti 6.3-MCS-IRES2-EGFP.After identification by sequencing , recombinant lentivirus was packaged by using pLenti 6.3-C3a-IRES2-EGFP and packaging plas-mid in 293T cells.Then, the recombinant lentivirus was used to in-fect HK2, a cell line of human renal tubular epithelial cells .After screening in medium with blasticidin , blasticidin resistant cell clones were obtained .Real-time PCR and ELISA method were applied to analyze the expression and secretion of stable transfected cells cloned C3a and identify renal tubular epithelial cell lines with stable over-activating C3a. Results ①C3a secretory expression unit was suc-cessfully synthesized and correctly cloned into the multi-clonal site of pLenti6.3-IRES2-EGFP; ②C3a secrectory expression recombi-nant lentivirus LV-C3a was successfully packaged with a high titer of 5 ×108/mL;③HK2 Cell clones resistant for blasticidin were ob-tained;according to the analysis of Real-time PCR and ELISA, the C3a mRNA level in HK2-C3a cell lines was significantly higher than that of HK2 cells(1.0 ±0.5 vs 1321.0 ±18.0, P<0.01) and the secreted C3a level increased significantly ([0.3 ±0.2]ng/mL vs [249.0 ±37.0] ng/mL, P<0.01). Conclusion The present study successfully constructed C 3a secretory expression vector pLenti6.3-C3a-IRES2-EGFP and C3a over-expression renal tubular epithelial cell line HK 2-C3a, which is very useful in further study of the function and significance of C 3a/C3aR axis not only in renal tubular epithelial cells but also in other cell types .
4.A study of urinary tryptase expression in patients with diabetic nephropathy and its pathological significance
Jingmin ZHENG ; Guang YIN ; Shifeng YUN ; Jianping WANG ; Zhihong LIU
Journal of Medical Postgraduates 2014;(11):1176-1179
Objective Increased renal mast cells have been found in significant correlation with clinicopathological features of diabetic nephropathy ( DN) .However, the exact pathological significance of mast cells still remained to be elucidated.As one of the most abundant serine proteases, tryptase is specifically expressed by mast cells.The study was to observe the expression of tryptase in the urine of patients with diabetic nephropathy and realize the pathological significance of urinary tryptase and renal mast cells in the development of diabetic nephropathy. Methods Urine samples from 103 patients with diabetic nephropathy who were hospitalized at the clinical unit of the nephrology centre of Jingling Hospital from January 2012 to June 2013 were collected.According to concentra-tion of urinary albumin excretion rate, the patients were conducted to early-stage DN group (microalbuminuria,n=10), middle-stage DN group (proteinuria stage, n=31) and end-stage DN group (renal insufficiency stage,n=62).Meanwhile, urine samples from 20 normal persons were taken as the normal control group.Tryptase level in each urine sample was measured by using an ELISA kit.The average tryptase levels were compared among different groups and the correlation between urinary tryptase level and clinical indices was analyzed. Results ①In most cases, tryptase was not detected in the urine samples from normal persons(0.7 ±1.4 ng/mg creati-nine).However, the urinary trypatse level increased significantly with the development of diabetic nephropathy (11.6 ±10.5 ng/mg creatinine for early stage, 29.7 ±19.2 ng/mg creatinine for middle stage, and 44.6 ±43.4 ng/mg creatinine for late stage group).②The urinary tryptase level was found in significant correlation with ser-um creatinine (r=0.325, P<0.01), serum cystatin C (r=0.391, P<0.01), serum urea nitrogen (r=0.27, P<0.01), 24 hour uri-nary protein (r=0.389, P<0.01), urine C3 (r=0.276, P<0.05), urine retinol-binding protein (r=0.365, P<0.01), urine lysozyme (r=0.461, P<0.01), urine N-acetyl-β-glucosaminidase level (r=0.568, P<0.01), urinary kidney injury molecule-1 (r=0.434, P<0.05) and interleukin-18 level (r=0.375, P<0.05).No significant correlation was found between urinary tryptase level and age, gender, fasting blood sugar, postprandial blood sugar, HbA1c, triglyceride, high density lipoprotein or low density lipo-protein. Conclusion Mast cells play an important role in the development of diabetic nephropathy and might be a novel and effective target for the treatment of DN.
5.Effect of nanosilver and nanohydroxyapatite mixed filling on osseointegration of immediate implants
Yu ZHENG ; Bingting LI ; Lijie WANG ; Shifeng FAN ; Xiaowei HOU
Chinese Journal of Tissue Engineering Research 2015;(47):7649-7654
BACKGROUND:Nanosilver has significant antibacterial properties, and nanohydroxyapatite has good biological activity and mechanical strength, while their mixture cannot only promote bone formation but also have antibacterial properties. OBJECTIVE:To observe the effect of nanosilver and nanohydroxyapatite mixed filing on the osseointegration of immediate implants. METHODS: Nine New Zealand white rabbits were randomly divided into experimental group (n=6) and control group (n=3) after four incisors from the upper and lower jaw were extracted. The mixture of nanosilver and nanohydroxyapatite was filed into the tooth socket in the experimental group, while nanohydroxyapatite was filed into the tooth socket in the control group. Titanium screw was immediately implanted into both groups. The intact maxila and mandibular specimens were harvested at the 4th, 8th, and 12th weeks after operation. Gross observation, X-ray bone density analysis, torque test and histological observation were conducted. RESULTS AND CONCLUSION:The gray value and maximum torque value of regenerated osseous tissue at different time points in experimental group were significantly higher than those in the control group (P < 0.05). Within 12 weeks of implantation, the bone formation rate and maturity of new bone tissue were higher in the experimental group compared with the control group, and no inflammatory cel infiltration occurred. At the 4th week after implantation, there was a large amount of inflammatory cel infiltration, and few inflammatory existed at the 8th week after implantation. These results demonstrate that compared with nanohydroxyapatite alone, the mixture of nanosilver and nanohydroxyapatite shows better antibacterial effect, biocompatibility and osteoinductive ability, which may accelerate osseointegration and promote osteogenesis.
6.Expression of DAB2IP and β-catenin in bladder urothelial carcinoma and their clinical significance
Linfeng ZHENG ; Xinghao NI ; Guoping CHENG ; Shifeng YANG
Chinese Journal of Clinical and Experimental Pathology 2017;33(4):398-402
Purpose To investigate the clinical utility of DAB2IP (DOC-2/DAB2 interactive protein)and β-catenin expression in bladder urothelial carcinoma (BUC).Methods The expression of DAB2IP and β-catenin was detected in 104 BUC cases and 40 peritumorial tissues using EnVision two-step immunohistochemical method,and the association with BUC clinicopathological parameters was analyzed.Results The expression of DAB2IP in BUC was significantly less than that of peritumorial normal tissues,and the expression of β-catenin in BUS was significantly higher than that of peritumorial normal tissues (P < 0.05).DAB2IP expression and histologic grading,clinical pathologic staging and 5 years survival rate had statistical significance (P < 0.05).No statistical significance with gender,age,tumor diameter and in patients with incipient/recurrence.β-catenin expression and age,histologic grading,clinical pathologic staging,tumor diameter and 5 years survival rate have statistical significance (P < 0.05).No statistical significance correlated with gender and in patients with incipient/ recurrence,DAB2IP and β-catenin expression in BUC are negatively correlated (P < 0.05).Conclusion In bladder urothelial carcinoma,down-regulation of DAB2IP and up-regulation of β-catenin,are in a negative correlation.Abnormal expression of DAB2IP and β-catenin is correlated with histologic grading,clinical pathologic staging and prognosis.
7.The local application of recombinant human parathyroid hormone (1-34) promotes osseointegration of an immediate implant
Zhihui SUN ; Yu ZHENG ; Lijie WANG ; Shifeng FAN ; Xiaowei HOU
Chinese Journal of Tissue Engineering Research 2017;21(26):4210-4216
BACKGROUND: Based on a mouse model of tibial implantation, some scholars have found that the CaP-coated implant with recombinant human parathyroid hormone (1-34) (PTH(1-34)) shows strong osteogenesis effect at early stage, but this coating has not been applied in the oral environment.OBJECTIVE: To explore the role of local application of PTH(1-34) on immediate implant osseointegration . METHODS: Nine New Zealand white rabbits were randomly divided into two groups (six in experimental group and three in control group). All of the tooth sockets were filled with heterogeneous freeze-dried bone firstly after four incisors of each rabbit were extracted. In the experimental group, a titanium screw with PTH(1-34) loaded CaP coating was implanted into each tooth socket, while in the control group, a titanium screw with only CaP coating was implanted. The animals were executed respectively at 4, 8, 12 weeks after operation, and the intact maxillary and mandibular specimens were harvested and tested by gross observation, bone density analysis, torque test, histologic al observation, X-ray observation.RESULTS AND CONCLUSION: The gray value and maximum torque value of regenerated osseous tissue at different time points in the experimental group were significantly higher than those in the control group (P < 0.05). Within 4-12 weeks after implantation, regenerated and mature bone tissue appeared earlier in the experimental group than the control group. A large amount of new blood vessels were seen in the experimental group at 8 weeks after implantation, while in the control group, there were only few new blood vessels. To conclude, the local application of PTH(1-34) can promote bone formation, improve the implant-bone bonding strength, and enhance the stability of the implant.
8.The relationship between NO and gossypol in decreasing sperm quality in male rats
Shifeng CHU ; Yuzhu WANG ; Lixiang ZHENG ; Juntian ZHANG
Chinese Pharmacological Bulletin 2003;0(11):-
Aim To elucidate the biochemical mechanism of gossypol in inducing the decline of sperm quality.Methods Gossypol was administered orally at the dose of 50 mg?kg-1/2 d for two weeks.Then,the sperm was collected from the left caudal epididymis and was analysed by CASA.The morphological changeand the concentration of nitric oxide(NO)in testes as well as the level of hormone〔follicle-stimulating hormone(FSH)〕,luteotrophic hormone(LH),testosterone(T)in serum were assayed.Results Gossypol could induce the decrease of sperm number and sperm quality.The concentration of NO in testes increased significantly.Among the three kind of hormone,only the concentration of T showed decrease after the oral administration of gossypol.NA1108 could antagonize the decline of sperm quality damaged by gossypol and decrease the content of NO in testes.Conclusions The concentration of NO in testes beyond normal value was one of the toxic mechanism of gossypol that contributed to the inhibition of spermatogenesis.Some drugs with the ability to reduce NO content in testes could also increase sperm quality.
9.Effect of continuous pressure on the proliferation of rabbit's bone marrow stem cells
Lei PENG ; Zheng SUN ; Zhen WANG ; Jun WANG ; Jianping BAI ; Shifeng SONG ; Zhao MA
Chinese Journal of Tissue Engineering Research 2009;13(27):5239-5242
BACKGROUND: Osteoblasts derived from bone marrow stromal stem cells are the main force of the cells In bone marrow cavity.In the artificial joint replacement cases, their life situation is closely related to conductive pressure of prosthesis.OBJECTIVE: To observe the continuous pressure on the proliferation of rabbit's bone marrow stromal stem cells, and to reveal the biomechanism of continuous pressure causing loosening and sinking of prosthesis and its activity after replacement.DESIGN, TIME AND SE'I-I'ING: A randomized study based on cytology was performed at the Institute of Othorpaedics, the Fourth Military Medical University of Chinese PLA from February 2004 to October 2007.MATERIALS: One New Zealand rabbit aging 1 month and weighing 0.5 kg was used to extract the bone marrow stromal calls.METHODS: The third-passaged bone marrow stromal stem cells were inoculated in seven 24-well culture plates at the density of 1 × 104 cells/well. After 14 hours, cells were adherent completely and cultured in DMEM culture media containing 10% fetal bovine serum. Thereafter, the four culture plates were randomly divided into 4 experimental groups, including control group without any pressure, and continuous pressure (20, 60, and 100 kPa) groups. The continuous pressure was performed for 3 hours every day.MAIN OUTCOME MEASURES: Absorbance was detected at day 1, 3, 5, and 7 after culture using MTT assay.RESULTS: Absorbance of the three pressure groups after 3, 5 and 7 days was significantly lower than the control group (P < 0.01). There was no significant difference in absorbance between 20 kPa continuous pressure group and 60 kPa continuous pressure group on the third day (P >0.05); but the absorbance was significantly higher than 100 kPa continuous pressure group (P < 0.05). On the fifth day, there was significant difference in absorbance between paired continuous pressure groups (P < 0.05),i.e., the higher the continuous pressure was, the lower the absorbance was. On the seventh day, the absorbance of the 100 kPa continuous pressure group was significantly lower than 20 kPa continuous pressure group (P < 0.05).CONCLUSION: Patients with eady post-joint replacement had better not stand on the ground; otherwise, a continuous stress will be caused so as to induce the restrain of bone marrow stromal stem calls in the bone marrow cavity, which is not helpful for bone healing and easily causes subsidence and loosening of prosthesis.
10.Research progress of role of SOCS-3 in treating liver fibrosis
Zili ZHANG ; Shifeng ZHAO ; Wenxuan XU ; Feng ZHANG ; Li WU ; Yin LU ; Shizhong ZHENG
Chinese Pharmacological Bulletin 2015;(12):1646-1651
Liver fibrosis occurs as compensatory responses to tis-sue repairing process in a wide range of chronic liver injures.It is characterized by excessive deposition of extracellular matrix (ECM)in liver tissues.The new discovery shows that suppres-sor of cytokine signaling (SOCS-3)is strictly associated with fi-brogenic progression of chronic liver diseases.Recent basic and clinical investigations also demonstrate that liver fibrogenesis is accompanied by SOCS-3,which critically determines the patho-
genesis and prognosis of liver fibrosis.This review summarizes current knowledge on SOCS-3 and the relationships between SOCS-3 and liver fibrosis.On the other hand,it also presents the different strategies that have been used in experimental mod-els to counteract excessive SOCS-3 and the role of SOCS-3 in the prevention and treatment of liver fibrosis.