0.05);the lung′s rale improvement was significant(diffe-)rent(?~2=4.75 P

Objective To prepare recombinant human prothrombin-2 expressed in Pichia pastoris, and assay the enzymatic and clotting activities of prothrombin-2 activated by prothrombin activator ecarin.Methods Human prothrombin-2 gene and Echis carinatus ecarin gene were synthesized separately on the basis of the cDNA sequences published in GenBank.The gene of prothrombin-2 was cloned into the expression vector pPICZαA.The expression vector pPICZαA/prothrombin-2 was transformed into glycoengineered P.pastoris, and then prothrombin-2 engineered P.pastoris was screened.The expression products were induced by methanol, purified by two-step chromatography and identified by diges-tion by PNGase F and analysis of pepetide fingerprint.The ecarin gene was cloned into the expression vector pcDNA3.1. The expression vector pcDNA3.1/Ecarin was transformed into HEK 293T cells and the culture supernatant of HEK 293T/Ecarin was collected.The reaction product of HEK 293T/Ecarin cell culture supernatant and purified prothrombin-2 was analyzed by S-2238,which was the chromogenic substrate for thrombin.Fibrinogen was used to measure blood clotting time. Results The purified protein of P.pastoris expressed prothrombin-2 culture supernatant was 37 ×103 .The relative molecular mass(Mr) of the purified protein was reduced to 35 ×103, which was consistent with the theoretical Mr of prothrombin-2 molecular weight.The purified protein was proved to be prothrombin-2 by peptide fingerprint identification. The purified prothrombin-2 processed by HEK 293T/Ecarin culture supernatant could hydrolyze S-2238 to produce yellow pNA, and D405 of pNA increased with the volume of the processed prothrombin-2 that could promote the plasma coagulation.The blood clotting time was close to that of the thrombin kit.Conclusion Prothrombin-2 is prepared by P.pastoris and activated toα-thrombin by ecarin.This technique may replace the method of extraction of prothrombin from plasma and can be used for the treatment of war wounds or for future clinical research.

Objective To explore the effects of benzirnidazole on spermatogenesis function and enzymatic activity in testis of male rats.Methods 40 male Wistar rats,clean degree,were randomly divided into four groups,10 in each.The rats in the low- dose group (20 mg/kg),the moderate-dose group (100 mg/kg) and the high-dose (200 mg/kg) were treated with benzimidazole at different doses by garage,once a day for 80 consecutive days.Simultaneously,the rats control group were treated with the equivalent volume of distilled water and tween-80.In the end of the experiment,the rats were weighed,the testis and epididymides were immediately excised and weighed,the appearance was observed,and the viscera coefficients of the bilateral testis and epididymides were calculated.The number and motility of sperms in the tail of epididymis,the activity of some enzymes (LDH, ACP,SDH,G-6-PDH) of testis were tested.Results The testis and epididymides in the control group and the low-dose group were pink,large,smooth and plump,but they were mauve,small,obviously atrophic in the moderate-dose group and the high-dose group. The viscera coefficient of the testis and epididymides,the number and motility of sperms in the tail of epididymis were significantly decreased in the moderate-dose group and the high-dose group (P

OBJECTIVETo observe the effects of blood activating wind dissipating acupuncture (BAWDA) on blood pressure (BP) of prehypertension (PHT) patients.

METHODSTotally 60 PHT patients were assigned to the control group and the acupuncture group according to random digit table, 30 in each group. All patients were intervened by life style. BAWDA was additionally performed in patients in the acupuncture group for 6 weeks (30 times). The improvement of BP after intervened by acupuncture was observed. BP success rates and the proportion of PHT progressing to hypertension (HT) were also observed after 6-week intervention of acupuncture and at 1-year follow-up.

RESULTSSystolic blood pressure (SBP) and diastolic blood pressure (DBP) decreased after 6-week intervention in the acupuncture. The BP control rate was 56.7% (17/30 cases) in the acupuncture group vs.10.0% (3/30 cases) in the control group with statistical difference (chi2 = 14.70, P < 0.01). At 1-year follow-up BP success rate was 36.7% (11/30 cases) in the acupuncture group, remarkably higher than that of the control group [13.3%, (4/30 cases)] (chi2 = 4.36, P < 0.05).

CONCLUSIONSBAWDA showed BP regulating roles in a gradually stable decreasing tendency. It also could elevate BP success rate of PHT, and reduce the risk of PHT progressing to HT.

Acupuncture ; methods ; Acupuncture Therapy ; methods ; Blood Pressure ; Humans ; Hypertension ; Prehypertension ; therapy ; Wind

AIM: To detect the effect of conjunction matrigel with mammary fad pat(MFP)implantation on the tumorigenesis, proliferation, apoptosis and metastasis of Her2 positive and negative breast cancer model. METHODS: The Her2 positive BT 474 and Her2 negative MDA-MB 231 breast cancer cells were injected into MFP of nude mice with or without matrigel to establish breast cancer model. The tumor volume was measured every 3 d. Followed up for 30 d after implantation, the nude mice were killed and the tumors and associated organs were dissected for pathological sectioning and staining with hematoxylin and eosin. The time of tumor formation and the tumorigenesis were determined after implantation. The tumor volume and metastasis rate were calculated and compared with each other. The proliferation and apoptosis of Her2 positive and negative tumors were also determined. RESULTS: Matrigel and MFP implantation technology shortened the time of tumorigenesis significantly(P<0.01). The tumorigenesis rate of BT 474 and MDA-MB 231 breast cancer cells did not show any different(P>0.05). The metastasis rate of MDA-MB 231 breast cancer cells were improved from 25.0% to 37.5%(P<0.05). CONCLUSION: Matrigel and MFP implantation can be combined to shorten the time of tumor formation by two kinds of breast cancer cells, and improve the metastasis of Her2 negative MDA-MB 231 cells. Using matrigel does not show any effect of proliferation and apoptosis on Her2 positive and negative breast cancer cells.

BACKGROUND: The quality of chest compressions can be significantly improved after training of rescuers according to the latest national guidelines of China. However, rescuers may be unable to maintain adequate compression or ventilation throughout a response of average emergency medical services because of increased rescuer fatigue. In the present study, we evaluated the performance of cardiopulmonary resuscitation (CPR) in training of military medical university students during a prolonged basic life support (BLS). METHODS: A 3-hour BLS training was given to 120 military medical university students. Six months after the training, 115 students performed single rescuer BLS on a manikin for 8 minutes. The qualities of chest compressions as well as ventilations were assessed. RESULTS: The average compression depth and rate were 53.7±5.3 mm and 135.1±15.7 compressions per minute respectively. The proportion of chest compressions with appropriate depth was 71.7％±28.4％. The average ventilation volume was 847.2±260.4 mL and the proportion of students with adequate ventilation was 63.5％. Compared with male students, significantly lower compression depth (46.7±4.8 vs. 54.6±4.8 mm,P<0.001) and adequate compression rate (35.5％±26.5％ vs. 76.1％±25.1％,P<0.001) were observed in female students. CONCLUSIONS: CPR was found to be related to gender, body weight, and body mass index of students in this study. The quality of chest compressions was well maintained in male students during 8 minutes of conventional CPR but declined rapidly in female students after 2 minutes according to the latest national guidelines. Physical fitness and rescuer fatigue did not affect the quality of ventilation.

OBJECTIVE: To observe ultrastructure changes of electrical injury in rats. METHODS: An experimental model of rats suffered from the low voltage were designed. Ultrastructure changes of electrical injured tissues were observed under transmission electron microscope. RESULTS: (1) Plasma of epithelium was concreted in the affected areas and inner membrane system was broken. (2) Hypercontraction bands were observed in skeleton muscles. (3) There was dissolved necrosis and hypercontraction bands in the myocardium. (4) Vacuoles were found in plasma of endothelium of blood vessels on electrical current path, and myelin sheath of nerve fiber were loosed. CONCLUSION The above mentioned ultrastructure changes could be used as assistant diagnostic index of electrocution. The mechanism of the changes were discussed.
Animals ; Electric Injuries/pathology* ; Muscle, Skeletal/ultrastructure* ; Nerve Fibers/ultrastructure* ; Rats ; Rats, Sprague-Dawley

To reduce the serum clearance of interferon alpha2b, a chimeric gene encoding an human serum albumin(HSA)--human interferon alpha2b(IFNalpha2b) fusion protein was overexpressed in Pichia pastoris. After fermentation in a 5L bioreactor, the fusion protein, capable of cross-reacting with anti-IFN alpha and anti-HSA antibody, was purified from the culture of the recombinant yeast by ultrafiltration, blue Sepharose affinity, phenyl hydrophobic interaction and Q ion exchange chromatography. Its IFNa2b moiety exhibits antiviral activity similar to that of recombinant human IFNa2b. In Cynomolgus monkeys model, The fusion protein was detectable in plasma, even 336h after a single does of 90 microg/kg injection intravenously or subcutaneously. The elimination phase half-life of the fusion protein was 101h after intravenous injection and 68.2h after subcutaneous injection. Its Subcutaneous bioavailability was 67.9%. The enhanced pharmacokinetics of interferon a2b fused to human serum albumin suggest its promissing application in clinic medicine.
Animals ; Bioreactors ; microbiology ; Fermentation ; Humans ; Interferon-alpha ; biosynthesis ; genetics ; Macaca fascicularis ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacokinetics ; Recombinant Proteins ; Serum Albumin ; biosynthesis ; genetics

In recent years, antibiotic resistance of bacteria has become a global health crisis. Especially, the new class of "superbug" was found in South Asia, which is resistant to almost known antibiotics and causes worldwide alarm. Through the underlying mechanisms of bacterial pathogenecity, the expression of many pathogen virulence factors is regulated by the process of quorum sensing. Screening efficient quorum sensing inhibitors is an especially compelling approach to the future treatment of bacterial infections and antibiotic resistance. This article focuses on bacterial quorum sensing system, quorum sensing screening model for in vitro and evaluation of animal models in vivo, recent research of quorum sensing inhibitors and so on.
Animals ; Anti-Bacterial Agents ; pharmacology ; therapeutic use ; Bacterial Infections ; drug therapy ; Disease Models, Animal ; Drug Resistance, Bacterial ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Pseudomonas aeruginosa ; drug effects ; pathogenicity ; physiology ; Quorum Sensing ; drug effects ; physiology ; Virulence ; drug effects ; Virulence Factors ; metabolism

OBJECTIVETo investigate the therapeutic effect of C-phycocyanin (C-PC) from Spirulina platensis on paraquat (PQ)-induced pulmonary fibrosis in rats.

METHODSA total of 90 healthy Wistar rats were randomly and equally divided into normal control group, model group (PQ group), and C-PC treatment group (C-PC group). Each rat in the PQ group and C-PC group were orally administered with a single dose of PQ (50 mg/kg) to establish a rat model of PQ poisoning. Then, the rats in the normal control group and PQ group were orally given saline solution (1 ml/100 g) every day, and the rats in the C-PC group were orally given C-PC (50 mg/kg) every day. Six rats were randomly selected from each group on days 1, 3, 7, 14, and 28. The inferior lobe of each rat's right lung was homogenized for the measurement of hydroxyproline (HYP) and maleic dialdehyde (MDA) levels and superoxide dismutase (SOD) activity. Parts of each rat's left lung were subject to HE staining and Masson staining for pathological observation, and the expression of transforming growth factor-β(1) (TGF-β(1)), nuclear factor-kappa B p65 (NF-κB p65), and tumor necrosis factor-α (TNF-α) in lung tissue was measured by immunohistochemistry.

RESULTSThe HYP levels on days 1, 3, 7, 14, and 28 and MDA levels on days 14 and 28 were significantly lower in the C-PC group than in the PQ group (P < 0.05, P < 0.01). The SOD activity was significantly higher in the C-PC group than in the PQ group on days 1, 7, 14, and 28 (P < 0.05, P < 0.01). The protein content of TGF-β(1) and the activities of NF-κB p65 and TNF-α in the PQ group and C-PC group were significantly higher than those in the normal control group, while the indices in the C-PC group were significantly lower than those in the PQ group (P < 0.05, P < 0.01). The pathological observation showed that C-PC could alleviate pulmonary alveolitis and fibrosis in rats with PQ poisoning.

CONCLUSIONC-PC can significantly inhibit PQ-induced pulmonary alveolitis and fibrosis in rats.

Animals ; Lung ; metabolism ; pathology ; Male ; NF-kappa B ; metabolism ; Paraquat ; poisoning ; Phycocyanin ; pharmacology ; Pulmonary Fibrosis ; chemically induced ; metabolism ; prevention & control ; Rats ; Rats, Wistar ; Transcription Factor RelA ; metabolism ; Transforming Growth Factor beta ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism