180,anti-ds-DNA,anti-Sm and lower C3,are high risk factors in the development of LN.The manifestations were various and misdiagnosis at the early stage was not uncommon.

OBJECTIVE: To construct a novel fusion protein contained insulin-like growth factor 1 (IGF-1) and lidamycin (LDM) and evaluate its antitumor activity on non-small cell lung cancer (NSCLC). METHODS: DNA fragment coding for fusion protein (ldp-igf) was synthesized by linking apoprotein of lidamycin (ldp) with igf-1, and then was cloned into the plasmid pET30a. Fusion protein LDP-IGF was expressed in E.coli as inclusion bodies and was purified by Ni2+ affinity chromatography. Binding affinity of LDP-IGF to NSCLC cells was evaluated by immunofluorescence assay and flow cytometry-based binding assay. MTT assay was used to measure the in vitrocytotoxicity of LDP-IGF and its enediyne-energized analogue LDP-IGF-AE. PI staining assay and Annexin V-FITC/PI staining assay were used to analyze the cell cycle arrest and cell apoptosis after treatment with LDP-IGF-AE, respectively. RESULTS: Active soluble LDP-IGF protein was prepared by isolation, purification, denaturation and refolding, and the production of LDP-IGF was 12 mg per liter fermentation broth. Both of immunofluorescence assay and flow cytometry-based binding assay showed that LDP-IGF has strong binding activity to NSCLC cells. Enediyne-energized fusion protein LDP-IGF-AE exhibited potent cytotoxicity to NSCLC cells in vitro, and it is more potent than that of LDM. Furthermore, fusion protein LDP-IGF without active enediyne was also cytotoxic to A549 cells at high concentrations (50 and 100 μg·mL-1). LDP-IGF-AE could cause significant G2-M arrest in A549 and H460 cells, and it also induced the apoptosis in NSCLC cells in a concentration-dependent manner. CONCLUSION: Fusion protein LDP-IGF-AE shows potent antitumor efficacy in vitro on NSCLC, suggesting it could be a promising candidate for targeted therapy.

OBJECTIVE: To construct a novel fusion protein consisting of oligopeptides specific for human epidermal growth factor receptor 2(HER2) and lidamycin(LDM), and investigate its antitumor activity. METHODS: Coding sequences of oligopeptides from complementarity determining region 3(CDR3) of anti-HER2 antibody C6.5 heavy chain was fused to apoprotein of lidamycin to obtain the fusion gene ldp-Hr. Fusion protein LDP-Hr was expressed in E. coli and purified by affinity chromatography. The purity of LDP-Hr was analyzed by high HPLC. Immunofluorescence assay and flow cytometry-based binding assay were used to investigate the binding activity of LDP-Hr to HER2 overexpressed cancer cells. The energized fusion protein LDP-Hr-AE was prepared by integrating the active enediyne chromophore (AE) of lidamycin into the LDP-Hr protein. MTT assay was used to measure the in vitro cytotoxicity of LDP-Hr-AE and Annexin V-FITC/PI staining assay was used to analyze its apoptosis-inducing efficacy. RESULTS: Fusion protein LDP-Hr was constructed correctly and expressed in E. coli in a secretory manner. The production of LDP-Hr was 40 mg per liter fermentation broth, and the purity of fusion protein was 97.4% as analyzed by HPLC. LDP-Hr showed strong binding activity to cancer cells highly expressing HER2, such as SK-BR-3 and SK-OV-3 cells. The energized fusion protein LDP-Hr-AE exhibited more potent cytotoxicity to SK-BR-3 and SK-OV-3 cells than LDM as measured by MTT assay. The results from Annexin V-FITC/PI staining assay also revealed that LDP-Hr-AE significantly induced cell apoptosis even at very low concentrations. CONCLUSION: The novel energized fusion protein LDP-Hr-AE bounds to HER2 specifically, and shows potent cytotoxicity and apoptosis-inducing activity to cancer cells, which suggests that it would be a promising candidate for targeted cancer therapy.

Forms and Records Control ; methods ; Humans ; Pathology, Clinical ; methods

Objective To explore the therapeutic effectiveness and pathways of human umbilical cord mesenchymal stem cells (hUCMSCs) transplantation for acute hepatic failure in rats.Method hUCMSCs were isolated from umbilical cord with attachment culture method,and the surface antigens were tested by flow cytometry.Forty-eight male Sprague Dawley rats were randomly divided into four groups.The animal model of acute liver failure was induced by injecting intraperitoneally with 50％ olive oil solution of carbon tetrachloride (2.5 ml/kg).The treatment groups were injected with hUCMSCs suspension separately through the tail vein or injected into the liver 24 h post-modeling.Blood serum and liver tissues were collected at several time points to analyze the improvement of liver function and histological repair.Real-time PCR was used to detect the expression of human CK8,CK18 and AFP mRNA in liver tissues.Immunohistochemistry was used to detect the expression of human CK18 in liver tissues.Result There were statistically significant differences among liver functions after transplantation (P＜0.05).hUCMSCs improved histological status through enhancing hepatocellular regeneration and reducing inflammatory cells.Real-time PCR results showed that the expression of CK8,CK18 and AFP mRNA was obviously increased in the tail vein transplantation group and hepatic lobe injection transplantation group as compared with the model group (P＜0.05).Immunochemistry results revealed that transplanted hUCMSCs in animal liver could differentiate into functional hepatocyte-like cells that expressed human CK18 as hepatocyte-specific marker in the tail vein transplantation group and hepatic lobe injection transplantation group.No significant differences in histological repair and grade of differentiation were examined between the tail vein transplantation group and hepatic lobe injection transplantation group (P＞0.05).Conclusion hUCMSCs can prompt the repair of acute liver failure and enhance pathological repair.Transplanted cells in animal liver can differentiate into functional hepatocyte-like cells that expressing hepatocyte-specific markers.Transplantation of hUCMSCs via the tail vein or direct injection into the liver had the similar therapeutic effects.

Objective To evaluate the methods for the prevention of internal jugular vein malposition of peripherally inserted central catheter (PICC) in patient with limited neck motion.Methods 210 patients who underwent PICC placement using ultrasound-guided modified Seldinger technique were divided into observation group (n =106) and control group (n =104) with a random number table.Ultrasound probe compression on the internal jugular vein was used in the observation group,while finger compression was used in the control group.The 2 groups were compared in terms of incidence of internal jugular vein malposition,accuracy of PICC tip position in X-ray,and incidence of complications.Results Incidence of PICC malposition was significantly lower in the observation group than the control group [3 (2.8％) vs.36 (34.6％),P =0.000].The accuracy of PICC tip position in both groups was 100％.No complication was observed in the observation group,while the rate of complication in the control group was 4.8％,with a statistically significant difference (P =0.022).Conclusion The ultrasound probe compression method can significantly lower the incidence of internal jugular vein malposition of PICC and is safer than the finger compression method.

Objective To construct the eukaryotic expression plasmid containing glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and cysteine protease inhibitor ( CPI ) gene from periodic Brugia malayi (Bm) and to lay foundation for studying multivalent vaccines. Methods Total RNA was extracted from periodic Bin. The BmGAPDH and BmCPI genes were amplified by RT-PCR. The PCR product was cloned and then subeloned into eukaryotic recombinant plasmid vector pcDNA3.1 (+). pcDNA3.1 (+)/BmGAPDH/BmCPI was constructed. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and were transformed into HeLa cell subsequently. The transient expression of BmGAPDH and BmCPI were examined by RT-PCR. The expressed protein was identified by sodium dodeeylsulphate-polyacrylamide gel electrophoresis(SDS-PAGE). Results Two specific bands of around 877 bp of BmGAPDH and 621 bp of BmCPI were amplified, consistent with the expected value. The same bands were obtained by double restriction enzyme digestion of recombinant plasmids or PCR using recombinant plasmid as template. BmGAPDH and BmCPI mRNA were highly expressed in transfeeted HeLa cell. The relative molecular mass (Mr) of the recombinant protein was about 54 × 103. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1 (+)/BmGAPDH/BmCPI has been constructed successfully and the protein is expressed correctly in mammalian cell.

Animals ; China ; Ehrlichia ; genetics ; isolation & purification ; Ehrlichiosis ; microbiology ; transmission ; RNA, Ribosomal, 16S ; genetics ; isolation & purification ; Sheep ; parasitology ; Ticks ; microbiology

Objective: To observe the clinical effects of Shu-acupuncture method in Nei Jing (Classic of Internal Medicine) in the treatment of shoulder and arm pain. Methods: A total of 90 patients with shoulder and arm pain were randomly divided into an observation group and a control group, 45 cases in each group. The treatment of Shu-acupuncture method in Nei Jing (Classic of Internal Medicine) was adopted in the observation group, routine acupuncture was used in the control group. The two groups were treated once every day, with 5 treatments as one course, and a 2-day rest between two courses. After 3 courses, pain was assessed by visual analog scale (VAS), and the clinical effects were compared between the two groups. Results: After the treatment, VAS scores were significantly changed in both groups (both P<0.01). The VAS score was lower in the observation group than that in the control group, with a statistical difference between the two groups (P<0.05). The total effective rate was 100% in the observation group, versus 91.1% in the control group, the difference was statistically significant (P<0.05). Conclusion: The therapeutic effect of Shu-acupuncture method in Nei Jing (Classic of Internal Medicine) is better than that of routine acupuncture in treating shoulder and arm pain.

In this study, hydrophilic matrix sustained release tablets of total lactones from Andrographis paniculata were prepared and the in vitro release behavior were also evaluated. The optimal prescription was achieved by studying the main factor of the type and amount of hydroxypropyl methylcellulose (HPMC) using single factor test and evaluating through cumulative release of three lactones. No burst drug release from the obtained matrix tablets was observed. Drug release sustained to 14 h. The release mechanism of three lactones from A. paniculata was accessed by zero-order, first-order, Higuchi and Peppas equation. The release behavior of total lactones from A. paniculata was better agreed with Higuchi model and the drug release from the tablets was controlled by degradation of the matrix. The preparation of hydrophilic matrix sustained release tablets of total lactones from A. paniculata with good performance of drug release was simple.
Andrographis ; chemistry ; Delayed-Action Preparations ; chemistry ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; Hydrophobic and Hydrophilic Interactions ; Kinetics ; Lactones ; chemistry ; Tablets ; chemistry