Surveillance of potential transmission factors of schistosomiasis in Xiuzhou District of Jiaxing City from 2013 to 2015 / 中国血吸虫病防治杂志

Objective To understand the potential risk of schistosomiasis transmission in Xiuzhou District of Jiaxing City, so as to provide the scientific evidence for consolidating schistosomiasis control achievements. Methods Fixed and mobile surveillance sites were set up in Xiuzhou District of Jiaxing City from 2013 to 2015. Oncomelania hupensis snails was surveyed historical snail habitats, current snail habitats, and suspected snail habitats. The schistosome infections were identified using serological and parasitological testing among local residents and mobile populations. In addition, the survival and reproduction of snails imported into Xiuzhou District was observed, and the schistosome infection in wild reservoir hosts was detected. Results A total of 540.14 hm2 of settings were surveyed in Xiuzhou District, Jiaxing City from 2013 to 2015, and 1.65 hm2 of snail habitats were identified. The snail habitats were mainly located in dry lands, and no infected snails or importation of snails were found. During the period from 2013 to 2015, a total of 7 668 local residents and mobile populations were examined in Xiuzhou District, and no new local infections were detected; however, three imported schistosomiasis cases were identified. Field simulation experiment showed that the imported snails laid eggs and reproduced in Xiuzhou District, and no schistosome infections were found in wild animals. Conclusion There are still residual Oncomelania snails and imported schistosomiasis patients in Xiuzhou District of Jiaxing City; therefore, the surveillance and management of local Oncomelania snails and imported schistosomiasis should be intensified to reduce the risk of schistosomiasis transmission.

Surveillance of potential transmission factors of schistosomiasis in Xiuzhou District of Jiaxing City from 2013 to 2015 / 中国血吸虫病防治杂志

Objective To understand the potential risk of schistosomiasis transmission in Xiuzhou District of Jiaxing City, so as to provide the scientific evidence for consolidating schistosomiasis control achievements. Methods Fixed and mobile surveillance sites were set up in Xiuzhou District of Jiaxing City from 2013 to 2015. Oncomelania hupensis snails was surveyed historical snail habitats, current snail habitats, and suspected snail habitats. The schistosome infections were identified using serological and parasitological testing among local residents and mobile populations. In addition, the survival and reproduction of snails imported into Xiuzhou District was observed, and the schistosome infection in wild reservoir hosts was detected. Results A total of 540.14 hm2 of settings were surveyed in Xiuzhou District, Jiaxing City from 2013 to 2015, and 1.65 hm2 of snail habitats were identified. The snail habitats were mainly located in dry lands, and no infected snails or importation of snails were found. During the period from 2013 to 2015, a total of 7 668 local residents and mobile populations were examined in Xiuzhou District, and no new local infections were detected; however, three imported schistosomiasis cases were identified. Field simulation experiment showed that the imported snails laid eggs and reproduced in Xiuzhou District, and no schistosome infections were found in wild animals. Conclusion There are still residual Oncomelania snails and imported schistosomiasis patients in Xiuzhou District of Jiaxing City; therefore, the surveillance and management of local Oncomelania snails and imported schistosomiasis should be intensified to reduce the risk of schistosomiasis transmission.

Early diagnosis of acute kidney injury in aged patients undergoing percutaneous coronary intervention / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

In aged patients, acute kidney injury (AKI) is a common clinical complication after percutaneous coronary intervention (PCI), highlighting the need for timely and certain diagnosis of this disease. A single centre, nested case-control study was conducted, which assessed the usefulness of urinary liver-type fatty acid-binding protein (uL-FABP), neutrophil gelatinase-associated lipocalin (uNGAL), and kidney injury molecule-1 (uKIM-1) for early detection of AKI. One hundred and thirty-two patients at or over 60 years old undergoing PCI were included. Serum creatinine (SCr) was measured before PCI, 24 and 48 h after PCI; uL-FABP, uNGAL, and uKIM-1 were measured before PCI, 6, 24, and 48 h after PCI. We identified 16 AKI patients and selected 32 control patients matched by admission time (<1 week), age (±5 years), and gender. In the receiver operating characteristic (ROC) curve analysis, the areas under the curve (AUCs) for the relative measurements of uL-FABP, uNGAL, and uKIM-1 were 0.809, 0.867, and 0.512 at 6 h after PCI, and 0.888, 0.840, and 0.676 at 24 h after PCI, respectively. AUC for the combination of uL-FABP and uNGAL was 0.899 at 6 h after PCI, and 0.917 at 24 h after PCI. Thus, measurement of uL-FABP and uNGAL levels at 6 and 24 h after PCI may be useful in detecting AKI in aged patients. Measurement of uKIM-1 levels provides inferior predictive power for early diagnosis of AKI.

Comparison of major bioactive components from leaves of Chrysanthemum morifolium / 中国中药杂志

Leaves of Chrysanthemum morifolium were potential medicinal resource. The present study aims to estimate the main bioactive components: total flavonoids (TF), galuteolin (GA), quercitrin (QU), chlorogenic acid (CA) and 3 ,5-O-caffeoylquinic acid ( CQ), which were considered to be the main effective components, in leaves of C. morfolium cultivars in China. The TF content was estimated hy UV-VIS spectrophotometry, while GA, QU, CA, and CQ were quantitatively determined by HPLC. The highest TF content (7. 13% w/w) was found in cultivar Wan Cong (Shexian county). Cultivar Da Bo ( Bozhou county) had the highest GA content (33. 45 mg - g-1); Cultivar Hong Xin (Sheyang county) contained the highest QU content (29.25 mg · g(-1)); Cultivar Chang Ban (Sheyang county) had the highest CA content (13.14 mg ·(-1)). The maximum CQ content (7.35 mg · g(-1)) was observed in culti- r Da Yang ( Tongxiang county). Different cultivars of C. morfolium had significant difference in components, but the leaf and capitulum of C. morifolium. were found to possess similar chemical compositions. The high content of bioactive components in several cultivars suggested the potential utilization of C. morifolium leaves.

Diagnosis value of non-enhanced magnetic resonance venography for deep pelvic vein disease / 第二军医大学学报

Objective To evaluate the diagnostic value of non-enhanced magnetic resonance venography (MRV) for deep pelvic vein disease. Methods A total of 50 patients highly suspicious of pelvic and lower extremity vein disease were enrolled in the present study, and they were subjected to lower extremity vascular 2D-TOF MRV(two-dimensional time-of-flight MR venography)examination with the following technical parameter: echo time 5-7 ms, repetition time 35-45 ms, and flip angle 35°-45°. The MRV range included the scanning from low segment of inferior vena cava (IVC) to the popliteal vein (PV); the image quality was scaled into grades, and the results of MRV were compared with those of ultrasound and DSA. Results The images of all 50 patients clearly showed the scanning from low segment of IVC to the PV and its branches, with the diagnostic accuracy reaching 96.0%. The images of 25 patients clearly showed a total of 723 veins, including IVC, common iliac vein (CIV), internal iliac vein, external iliac vein (EIV), common femoral vein, deep femoral vein, superficial femoral vein and PV, with a consistent rate of 96.4%. Thrombosis from inferior segment of IVC to EIV was shown on MRV images of 9 patients, while it could not be clearly and completely manifested by B-ultrasound. Ten patients received DSA simultaneously, and the MRV results of 9 were in accord with those of DSA findings. MRV of one patient with thrombosis at initial segment of CIV was shown normal on DSA. Conclusion MRV for diagnosis of lower extremity vascular lesions has the advantage of non-trauma, greater scanning range, high grade contrast, excellent image delineation and intuitive convincement, making it worth popularizing in clinic.

Inhibitory effects of anti-gelatinases scFv and lidamycin fusion protein on proliferation and metastasis of human fibrosarcoma HT-1080 / 中国药学杂志

OBJECTIVE: To investigate the antitumor efficacy of anti-gelatinases dFv-LDP and its enediyne-energized fusion protein dFv-LDP-AE on human fibrosarcoma HT-1080 cancer cells. METHODS: Western blot was used to analyze the expression level of gelatinases in different cancer cell lines. The inhibitory effects of fusion protein dFv-LDP and its enediyne-energized fusion protein dFv-LDP-AE on HT-1080 were determined by MTT assay. The binding capability of fusion protein dFv-LDP with HT-1080 was detected by ELISA and immunofluorescence. FACS was used to analyze the cell cycle arrest by dFv-LDP, dFv-LDP-AE or their combination on HT-1080 cells. The anti-metastasis effects of dFv-LDP, dFv-LDP-AE or their combination on the experimental lung metastasis model established by HT-1080Luc via tail vein injection were also evaluated in this study. RESULTS: Expression level of gelatinases was higher in HT-1080 cells as compared to that of other cancer cell lines. The fusion protein dFv-LDP showed well binding capability with HT-1080 cells as determined by ELISA and immunofluorescence. The enediyne-energized fusion protein dFv-LDP-AE displayed extremely inhibitory effect on proliferation of HT-1080. Results of FACS indicated that the combination of dFv-LDP with dFv-LDP-AE could not further increase the G2/M proportion on cell cycle arrest. However, in vivo experiment as examined using the experimental lung metastasis model established via HT-1080Luc tail veil injection, the metastasis foci in group of fusion protein dFv-LDP (10 mg·g-1) was 55.8% compared to that of control group (P<0.01). The metastasis foci in group of dFv-LDP-AE at dosage of 0.4 and 0.6 mg·g-1 were 41.4% and 25.1% respectively compared to that of dFv-LDP 10 mg·g-1 group (P<0.01). The combination of dFv-LDP (10 mg·g-1) with dFv-LDP-AE (0.4 or 0.6 mg·g-1) showed an additive decrease of metastasis foci number in the lung of athymic mice, which were 20.3% (P<0.05, compared with dFv-LDP-AE at 0.4 mg·g-1) and 13.1% (P<0.05, compared with dFv-LDP-AE at 0.6 mg·g-1) respectively. CONCLUSION: The combination of dFv-LDP with its enediyne-energized fusion protein dFv-LDP-AE would intensify the anti-metastasis effect on experimental lung metastasis model as established via tail vein injection of HT-1080Luc cells.

Bioluminescence imaging evaluation of the inhibitory effect of lidamycin on lung metastasis of human fibrosarcoma in athymic mice / 药学学报

This study is to investigate the inhibitory effect of lidamycin (LDM) and its combination with methotrexate (MTX) on lung metastasis of fibrosarcoma by bioluminescence imaging in athymic mice. A stable luciferase transfected HT-1080 cell line was constructed and the capability to establish experimental lung metastasis in athymic mice was confirmed. The optical imaging system was applied to evaluate the formation of lung metastasis in vivo. In addition, metastatic nodules were counted for the evaluation of inhibition rates. As shown, the fluorescent intensity of luciferase-transfected HT-1080 cells was colinear with the cell population and the minimal detected cell population was 100 cells/well. Optical imaging showed that the fluorescent intensity of treated group was apparently lower than that of the control. The inhibition rates of lung metastasis by LDM alone at 0.025 mg x kg(-1) and 0.05 mg x kg(-1) were 53.9% and 75.9%, respectively, while that of MTX alone at 0.5 mg x kg(-1) was 70.2%. The combination of LDM at 0.025 mg x kg(-1) and MTX at 0.5 mg x kg(-1) showed an inhibition rate of 88.7%. The coefficient of drug interaction (CDI) was 0.82. The results herein demonstrated that LDM alone had strong anti-metastasis effect on human fibrosarcoma HT-1080 and the inhibition efficacy is strengthened when combined with MTX.

Optimization of the assembly efficiency for lidamycin chromophore bound to its apoprotein: a case study using orthogonal array / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Lidamycin (LDM) can be dissociated to an apoprotein (LDP) and an active enediyne chromophore (AE). The detached AE can reassemble with its LDP-containing fusion protein to endow the latter with potent antitumor activity. However, the reassembly of AE with LDP is affected by several factors. Our aim was to optimize the assembly efficiency of the AE with a LDP-containing fusion protein and investigate the influence of several factors on the assembly efficacy.</p><p><b>METHODS</b>A method based on RP-HPLC was developed to analyze the assembly rate, and an orthogonal experimental design L(9) (3(4)) was used to investigate the effects of temperature, assembly time, pH and molecular ratio of LDP-containing fusion protein to AE on the assembly rate. Furthermore, the determined optimum conditions for the assembly rate of the LDP-containing fusion protein with AE were applied and evaluated.</p><p><b>RESULTS</b>A calibration curve based on the LDM micromolar concentration against the peak-area of AE by HPLC was obtained. The order in which individual factors in the orthogonal experiment affected the assembly rate were temperature>time>pH>molar ratio of AE to protein and all were statistically significant (P<0.01). The optimal assembly conditions were temperature at 10°C, time of 12 h, pH 7.0, and the molar ratio of AE: protein of 5:1. The assembly rate of AE with a LDP-containing fusion protein was improved by 23% after condition optimization.</p><p><b>CONCLUSION</b>The assembly rate of chromophore of lidamycin with its LDP-containing fusion protein was improved after condition optimization by orthogonal design, and the optimal conditions described herein should prove useful for the development of this type of LDP-containing fusion protein.</p>

Epidemiological effects of the A(H1N1)influenza vaccine immunization program on students / 中华流行病学杂志

Objective To evaluate the epidemiological effects of vaccine immunization program related to A(H1N1)influenza in the middle school students.Methods Non-randomized clinical trial was designed to assess the A(H1N1)influenza vaccine on its efficacy.14883 students from 8 middle schools in Zhejiang province were recruited and classified into vaccinated or control groups,based on the status of immunization with A(H1N1)influenza vaccine.All subjects were followed up through one epidemic period(6 months)and the incidence rates of influenza-like illnesses,A(H1N1)influenza,and seasonal influenza in these two groups were compared to evaluate the efficacy of the vaccine.Results There were 6334 subjects in the vaccinated group and 8549 in the control group.7441.75 person-years were followed from these two groups.The incidence rate of A (H1N1)influenza in vaccinated group was 1.64‰ per person-year,lower than that of the control group.The rate difference(RD)was-1.64‰ per person-year(95% confidence interval value from-3.04‰ to-0.23‰ per person-year),and the difference was significant(P=0.010).The incidence rate of influenza-like illnesses in vaccinated group was 21.47‰ per person-year,lower than that of the control group(22.69‰ per person-year)and the diffefence was not significant(P>0.05).The incidence rate of B influenza in vaccinated group was 6.63‰ per person-year,higher than that of control group(7.02‰ per person-year)but the difference was not significant(P>0.05).Conclusion This vaccine demonstrated a good epidemiological effect against the A(H1N1)influenza virus infection,observed through a student-immunization program.The cross-protection effect against the influenza-like illnesses and other seasonal influenzas was not noticed in this study.

Establishment of loop-mediated isothermal amplification method for detection of Legionella pneumophila / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a simple and rapid molecular detection for Legionella pneumophila.</p><p><b>METHODS</b>The loop-mediated isothermal amplification (LAMP) was applied for detection of Legionella pneumophila. A set of primers were designed to identify six special areas in mip gene of Legionella pneumophila. Genomic DNAs from 13 bacterial strains,including 8 Legionella pneumophila strains and 5 other bacterial strains were amplified by LAMP and general PCR method to evaluate the specificity and sensibility of LAMP.</p><p><b>RESULT</b>All positive tubes produced visible white precipitation, and no precipitation was observed in others. By adding smart green fluorescent dye, all Legionella pneumophila positive tubes presented a strong green fluorescence, while others showed weak fluorescence. The detection rate of LAMP was higher than that of general PCR. The detection limits were 576fg with genomic DNA of Legionella pneumophila,and 8 cfu/mL with positive water samples.</p><p><b>CONCLUSION</b>LAMP detection of Legionella pneumophila is an effective and low-cost method with high specificity and sensitivity requiring no special equipment.</p>

Preparation and identification of a polyclonal antibody against novel cysteinyl leukotriene receptor GPR17 / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To prepare and identify a polyclonal antibody (pAb) against GPR17, a novel cysteinyl leukotriene receptor.</p><p><b>METHODS</b>Rabbits were immunized with KLH-coupled GPR17 peptide to prepare the pAb. The titer of the pAb in rabbit plasma was detected by indirect ELISA, and the specificity of the pAb was tested by antigen blockade. GPR17 tissue distribution was detected by Western blot with the pAb.</p><p><b>RESULTS</b>The pAb showed a titer as high as 1:16 364,and was not cross-reacted with the antigens of CysLT(1) and CysLT(2) receptors. A higher expression of GPR17 in the rat brain and heart was detected using the newly prepared pAb. The molecular weigh of GPR17 protein was about 43 kD.</p><p><b>CONCLUSION</b>The prepared GPR17 pAb has high sensitivity and specificity,and can be used in Western blot for detecting GPR17.</p>

Preparation and identification of polyclonal antibody against cysteinyl leukotriene receptor 2 / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To prepare and identify a polyclonal antibody against cysteinyl leukotriene receptor (CysLT(2)receptor).</p><p><b>METHODS</b>Rabbits were immunized with KLH-coupled CysLT(2) receptor peptide to prepare the polyclonal antibody (pAb). The titer of the pAb in rabbit plasma was detected by indirect ELISA, and the specificity of the pAb was tested by antigen blockade. The tissue distribution of CysLT(2) receptor was detected by Western blot and immunohistochemistry with the prepared pAb.</p><p><b>RESULT</b>The pAb showed a titer higher than 1/1047296, and was specific to CysLT(2) receptor, without cross-reaction with the antigens of CysLT(1) receptor and GPR17. A higher expression of CysLT(2) receptor in kidney, brain and lung of rats and mice was detected by Western blot analysis using the prepared pAb. The molecular weight of CysLT(2) receptor protein was about 40 kD. Immunohistochemical examination showed that CysLT(2) receptor was expressed mainly in the neuron, and partly in astrocytes in rat brain.</p><p><b>CONCLUSION</b>The prepared CysLT(2) receptor pAb has high sensitivity and specificity, and can be used in Western blot and immunohistochemistry.</p>

Method for screening cysteinyl leukotriene receptor 2 antagonists and preliminary screening of compounds / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a method for screening cysteinyl leukotriene receptor 2 (CysLT(2)) antagonists and to preliminarily screen a series of synthetic compounds.</p><p><b>METHODS</b>Rat glioma cell line (C6 cells) highly expressing CysLT(2) receptor was used. Intracellular calcium concentration was measured after stimulation with the agonist LTD(4),which was used to screen compounds with antagonist activity for CysLT(2) receptor. Bay u9773, a CysLT1/CysLT(2) receptor non-selective antagonist, and AP-100984, a CysLT(2) receptor antagonist, were used as control.</p><p><b>RESULT</b>PT-PCR showed a higher expression of CysLT(2) receptor in C6 cells. LTD(4) at 1 mumol/L significantly increased intracellular calcium in C6 cells; the maximal effect was about 37.5% of ATP, a positive stimulus.LTD(4)-induced increase of intracellular calcium was blocked by CysLT(2) receptor antagonists, but not by CysLT(1) receptor antagonists. Among the synthetic compounds, D(XW-)1,2,13,23,29 and 30 inhibited LTD(4)-induced increase of intracellular calcium.</p><p><b>CONCLUSION</b>LTD(4)-induced change in intracellular calcium in C6 cells can be used as a screening method for CysLT(2) receptor antagonists. The compounds, D(XW-)1,2,13,23,29 and 30, possess antagonist activity for CysLT(2) receptor.</p>

Expression of tyrosine phosphatase containing C-src homology SH-2 in benign prostate hyperplasia / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To explore the expression of tyrosine phosphatase containing C-src homology SH-2 (SHP-1 and SHP-2) in benign prostate hyperplasia.</p><p><b>METHODS</b>With En Vision two-step method, the expression of SHP-1 and SHP-2 was detected in 10 cases of normal prostate tissue, 30 cases of BPH, 20 cases of PIN, 20 cases of high differential Pca and 20 cases of low differential Pca.</p><p><b>RESULT</b>The expression of SHP-2 in normal group was mainly distributed in the cytoplasm of secretive cells and basal cells, and a little part in the nucleu. In BPH it was distributed equally in the plasm and nucleu. In PIN, high differential Pca and low differential Pca, SHP-2 expressed mainly in nucleu. The average dyeing index of SHP-2 in each group is 0.4, 1.7, 2.1, 2.2 and 2.6. SHP-1 positive expression in normal prostate, BPH, PIN and high differential Pca showed differentiating layer staining in the cytoplasm of secretive cells and basal cells, while not in low differential Pca. The average dyeing index of SHP-1 in each group is 1.8, 1.8, 1.5, 1.2 and 0.4.</p><p><b>CONCLUSION</b>There are transformation in signal transduction relation with SHP-1 and SHP-2 in the progress of prostate cell proliferation, differentiation and malignant. The abnormal activation and distribution of SHP-2 might induce prostate reconstruction and hyperplasia, even carcinoma.</p>

Comparative study on inner quality of various Chrysanthemum morifolium cultivated in Tongxiang city / 中国中药杂志

<p><b>OBJECTIVE</b>To study the quality and the chemical components of Chrysanthemum morifolium from Tongxiang city.</p><p><b>METHOD</b>Chemical constituents of nine cultivars were compared in three types of index: chlorogenic acid, flavonoid and volatile oil.</p><p><b>RESULT AND CONCLUSION</b>The content varied significantly. The content of chlorogenic acid in Jinjuerhao was 6.66%, the highest among the samples. Yizhongdabaiju showed the highest flavonoid and volatile oil with 9.49% and 3.30 mL x kg(-1) respectively.</p>

Adhesional inhibiton of polyclonal anti-porin I antibody / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the blockness effects of purified polyclonal anti-porin I antibody on N. gonorrhoeae adherence to genitourinary tract epithelia of BALB/c mouse.</p><p><b>METHODS</b>Polyclonal anti GST-PI antibody was generated by immunizing rabbit with GST-PI fusion protein which was constructed and expressed by ourselves. The purified immunoglobulin G was obtained by ammonium sulphate deposition and DEAE cellulose chromatography. Mice model of gonorrhea was established. In order to evaluate the effects of PI-IgG on gonococcus adhesion to vagina mucus, the macroscopic and pathological assessing as well as gonococcus culture was employed after gonococcus challenge on PI-IgG immunized mice.</p><p><b>RESULT</b>No pus and pathological inflammation were observed on mice vagina mucus treated with 1 mg/ml PI-IgG 3 hours before gonococcus challenge. Gonococcus could not be detected in the smears and washing solutions from vagina. Pathological inflammation was found in mice treated with anti PI-IgG, in which the concentrations were lower than 1 mg/ml or the treated time was longer than 3 hours prior to gonococcus challenge.</p><p><b>CONCLUSION</b>The purified anti PI-IgG can effectively inhibit the adherence and infection of gonococci to genitourinary tract epithelia of BALB/c mice. In addition, the blocking duration of anti PI-IgG is associated with antibody concentration.</p>

Purification of Recombinant Fusion Protein Staphylokinase-Hirudin Expressed by Escherichia coli and Analysis of its Dimer / 中国生物工程杂志

The recombinant fusion protein staphylokinase-hirudin(rSFH) was purified from the high density-fermented engineered E.coli by means of ion-exchange chromatography (IEC) and gel filtration (GF). The purity of rSFH reached to more than 98% determined by RP-HPLC and SDS-PAGE, and the yield was up to 0.7g per liter of fermentation broth. The analysis of homologous dimmer of rSFH appeared during the purification and calculation of the surface hydrophobic area had been carried out by means of hydrophobic chromatography and MALD-TOF. The influence of sodium chloride and temperature on the behavior of rSFH reversible dimerization was analyzed by high performance sized- exclusive chromatography(HPSEC). It is concluded that the hydrophobic interaction played an important role in the reversible dimerization of rSFH.

Experiment treatment of collagen-induced arthritis in rats with recombinant plasmid containing vasoactive intestinal peptide gene / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the therapeutical effect of recombinant plasmid containing vasoactive intestinal peptide gene (pcDNA3.1+/VIP) on collagen-induced arthritis (CIA) in rats.</p><p><b>METHODS</b>The experimental arthritis was induced by intradermal injection of bovine type II collagen emulsified in Freund's adjuvants in male SD rats. The rats then were given intra-articular injection with recombinant plasmid (pcDNA3.1+/VIP). The levels of serum TNF-alpha, IL-4 and IL-2 were detected by Avidin-Biotin Peroxdase Complex-enzyme-linked immunosorbent assay (ABC-ELISA) and the pathological changes in the joint of rats were observed.</p><p><b>RESULT</b>Histological examination showed massive inflammatory infiltration in the joint with destruction of bone and cartilage, while the severity of pathological changes in synovia of VIP-treated rats was markedly reduced. Compared with normal group, the serum TNF-alpha, IL-2 levels of CIA rats were significantly increased (P <0.05) and IL-4 level was decreased (P<0.05). Compared with control and pcDNA3.1+ -treated CIA rats, serum TNF-alpha and IL-2 levels of pcDNA3.1+/VIP-treated rats were decreased and IL-4 level was increased (P<0.05).</p><p><b>CONCLUSION</b>Recombinant plasmid containing vasoactive intestinal peptide gene (pcDNA3.1+/VIP) can reduce the clinical and histological severity of established CIA and it might be a promising candidate for treatment of rheumatoid arthritis.</p>

Preparation of monoclonal antibodies against human mesenchymal stem cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To prepare monoclonal antibodies (McAbs) against human mesenchymal stem cells (hMSCs) and to study their biological characteristics.</p><p><b>METHODS</b>BALB/C mice were immunized with pooled hMSCs. McAbs were prepared by hybridoma technique and their biological characteristics were analyzed by indirect immunofluorescence, immunohistochemistry and flow cytometry.</p><p><b>RESULT</b>Five hybridoma cell lines were successfully established, which secret McAbs specifically against hMSCs. Investigations showed that all these McAb reacted only to hMSCs and had no cross-reaction to other human cells, the relative affinities of 5 McAbs were 1x10(6) (ZUB1), 1x10(5) (ZUB4), 1x10(6) (ZUC3), ZUE12 (1x10(5)) and 1x10(5) (ZUF10), respectively. Isotype analysis showed that ZUB1, ZUE12, ZUF10 against the same isotype, while ZUC3, ZUB4 against other two different isotypes alone. Flow cytometric analysis showed that the positive expression rate of cultured hMSCs was 87.39% (ZUB1), 88.07% (ZUB4), 88.12% (ZUC3), 69.89% (ZUE12) and 83.67% (ZUF10).</p><p><b>CONCLUSION</b>The prepared five McAbs can specifically react against hMSCs, which can be used for selection and study of hMCSs.</p>