AIM:To separate and purify a new anti-clotting protein component(ACPF-7221)from the venom of Agkistrodon. The acute toxicity experiment of ACPF-7221 and the death rate were studied and the LD50 and 95% confidence interval were determined in mice. METHODS: The venom of Agkistrodon was separated and purified using ion-exchange chromatography of DEAE-Sepharose Fast Flow, molecular sieve filtration through Sephadex G75, SP-Sepharose Fast Flow and molecular sieve filtration through Sephadex G50, the anticoagulation components were recalled using activated partial thromboplastin time (APTT) activity assay. ACPF-7221 was given to mice by intravenous injection,and the dosage was determined according to the body weight of mice. The LD50 and 95% confidence were calculated by the death outcome. RESULTS: ACPF-7221 was separated and purified,which was a trimer consisted of three kinds of protein and their relative molecular mass were 25000, 30000 and 50000,and the protein level was 65%. The LD50 of ACPF-7221 was 7.848 mg/kg, and its 95% confident interval was 7.353-8.375 mg/kg. CONCLUSION: A new anti-clotting protein component ACPF-7221 was extracted from the venom of Agkistrodon by ion-exchange chromatography,which is a trimer consisted of three kinds of protein. The critical orga of acute toxicity is lung, empsyxis asphyxia deduces death.

Communicable Disease Control ; Humans ; Research

Objective: To investigate the effects of overexpression of microRNA-7 (miR-7) on the proliferation and invasion of HepG2 cells and the underlying mechanism in vitro. Methods: The relative expression levels of miR-7 and Raf1 in hepatocellular carcinoma (HCC) tissues and adjacent normal tissues (ANT) were detected by quantitative real time-PCR (qRT-PCR). The relationship between the expression of miR-7 and the characteristics of HCC patients was analyzed. Cells were divided into blank control group, negative control (NC) group and miR-7 mimics transfected group, miR-7 mimics and NC were transfected into HepG2 cells by Lipofectamine™2000. The relative expression of miR-7 was detected by qRT-PCR. The proliferation ability of HepG2 cells was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The invasion of HepG2 cells was detected by Transwell assay. The target genes of miR-7 were predicted by TargetScan and the binding effect of miR-7 on the 3′UTR of Raf1 was verified by dual luciferase reporter assay.The expressions of Raf1 protein in hepatocellular carcinoma tissues, normal tissues and miR-7 mimics transfected HepG2 cells was detected by Western blot. The correlation of the levels of miR-7 and Raf1 mRNA was determined by Pearson correlation analysis. Results: The relative expression level of miR-7 in HCC was 0.49±0.02, significantly lower than in ANT (1.21±0.05, P<0.01). The level of miR-7 was significantly correlated the tumor volume, metastasis and prognosis of HCC patients (P<0.05). The relative expression level of miR-7 in miR-7 mimics transfected HepG2 group was 12.67±0.40, significantly higher than that in blank group (P<0.01). Compared with the blank group, the A value and invasion ability of miR-7 mimics transfected group were significantly down-regulated at 48 hours and 72 hours after transfection (P<0.01). Compared with miR-7 NC group, the luciferase activity of wild-type Raf1 reporter gene in miR-7 mimics transfected group was significantly reduced (P<0.01). The relative expression of Raf1 protein in HCC was 3.15±0.10, significant higher than in ANT (0.53±0.03, P<0.01). The relative expression of Raf1 protein in miR-7 mimics transfected group was 0.24±0.01, significantly lower than in miR-7 NC group (0.98±0.02, P<0.01). Furthermore, an negative correlation was observed between the levels of miR-7 and Raf1 in HCC tissues (P<0.05). Conclusions The expression of miR-7 in HCC is significantly decreased and inversely correlated with poor survival of HCC patients. Overexpression of miR-7 can inhibit the proliferation and invasion ability of hepatocellular carcinoma cells HepG2 by downregulating Raf1 in vitro.

Objective To study the effects of hyperbaric oxygen (HBO) preconditioning (PC) on motor function and mitochondrial apoptosis after spinal cord injury (SCI) in rats.Methods Thirty-six healthy,male Wistar rats were randomly divided into 3 groups (each n =12):a control (CON) group,an SCI model group and an HBO-PC group.The SCI group and the HBO-PC group after 7 sessions of HBO-PC treatment were subjected to SCI modeling using Allen's method.The CON group was not given any special treatment.Two weeks after the modeling,Basso,Beattie,and Bresnahan BBB scale was used to rate the rats' locomotor function,and the T8 segment of the rats' spinal cords was removed.After extracting total RNA from the spinal cord tissue,real-time quantitative PCR was used to detect the mRNA expression levels ofcaspase-3,cytochrome C (CYC),Bak,Bax,Bcl-x and Bcl-2 in the spinal cord tissue.Results Two weeks after modeling,compared with the CON group,the average BBB locomotion score of the SCI model group had decreased.In contrast,caspase-3,CYC,Bak and Bax mRNA expressions had increased significantly.Compared with the CON group,the average BBB locomotion score of the HBO-PC group had decreased,caspase-3 expression had increased,and Bax mRNA expression had decreased,but the differences in the changes between the two groups were not statistically significant.Bak mRNA expression increased and CYC expression decreased,and the difference between the two groups was statistically significant.Compared with the SCI model group,the average BBB locomotion score of the HBO-PC group increased while caspase-3,CYC,Bak and Bax mRNA expressions decreased,but none of these differences between the two groups was statistically significant.Bak mRNA expression increased and CYC mRNA expression decreased,and those differences between the two groups were statistically significant.Bcl-x and Bcl-2 mRNA expression in the CON,HBO-PC and SCI model groups were significantly different.Conclusion HBO-PC can reduce the loss of motor function after SCI,at least in rats.The mechanism may be related to reducing neuron mitochondrial apoptosis.

Objective To compare the therapeutic effects of hyperbaric oxygen (HBO) therapy at different time points on patients with spinal cord injury. Methods A total of 284 cases of spinal cord injury (SCI) were divided into an HBO group and a control group. The control group was treated with routine interventions (including a dehydrator, medication for neurotrophy, rehabilitation management, acupuncture and supportive treatment). The HBO group was treated with HBO in addition to the routine treatments, initiated at different time points after onset of SCI ( from less than 8 hours to longer than 1 week). All of the patients were evaluated in terms of scores on the American Spinal Cord Injury Association (ASIA) assessment and Barthel's index (BI) before and after treatment.Results Both groups had excellent effects when the treatment was initiated within the first 8 hours after the onset of SCI. There was no significant difference between the two groups. Average ASIA and BI scores in both groups improved significantly within the first 24 hours, and over the 1 st week after the onset of SCI. The recovery of neurological function and ability in the activities of daily living (ADL) was significantly better when HBO treatment was initiated within 8 hours after SCI compared with that initiated after 24 hour and 1 week. The therapeutic effects in the HBO group patients were significantly better than in the control group when HBO was initiated at 8 and 24 hours after onset of SCI. Neither the HBO group nor the control group had significant further improvement in ASIA scores or ADL performance when treatments initiated after 1 week. Conclusions HBO can significantly improve neurological function and ADL performance after SCI if it is administered within 1-8 hours.

Animals ; China ; epidemiology ; Humans ; Risk ; Streptococcal Infections ; epidemiology ; microbiology ; pathology ; prevention & control ; Streptococcus suis ; classification ; isolation & purification ; metabolism ; pathogenicity ; Virulence Factors ; metabolism

AIM: To observe the efficacy of ondansetron by intravenous injection at different time in preventing nausea and vomiting after pediatric strabismus surgery.METHODS: Totally 90 children aged 3 - 11y were randomly selected for pediatric strabismus surgery from June 2013 to August 2013 in our hospital. The ASA grade of all children were Ⅰ - II. Children were randomly divided into three groups with 30 cases each. Group A received intravenous drip of ondansetron 0. 1mg/kg before surgery. Group B received intravenous drip of ondansetron 0. 1mg/kg after surgery. Group C as control group was not given ondansetron. The number and severity of nausea and vomiting were observed within 24h after surgery.
RESULTS: There were no statistical significance in patients' gender, weight, age, duration of anesthesia, ketamine dosage and vital signs intraoperative between the three groups ( P > 0. 05 ). The incidence rate of postoperative nausea and vomiting ( PONV) of group A and B were significantly lower than group C (P<0. 05). The incidence rate of PONV of group A and group B have no significant difference (P>0. 05).
CONCLUSION:Using ondansetron is effective and safe in preventing PONV before and at the end of the pediatric strabismus surgery, which can also improve safety and be lower cost. It is a worthy promoting antiemetic approach for eye surgery.

Objective The epidemiological characteristics and risk factors of infection of non-tuberculous mycobacterial (NTM) by elderly bronchiectasis patients in Huai 'an area were analyzed, and the theoretical basis for prevention of NTM infection by elderly bronchiectasis patients in Huai 'an area was provided. Methods Among the 371 elderly patients with bronchiectasis admitted to our hospital from January 2020 to June 2022 were selected and divided into control group and NTM group according to whether they had NTM or not. The NTM strains were isolated and identified. Clinical data of patients were collected from the medical record system. Independent risk factors of NTM infection in elderly patients with ramus were analyzed by univariate analysis and logistic regression, including gender, age, previous smoking status, number of ramus, pulmonary cavities, hypoproteinemia, and CD4+T cell level. Results A total of 108 cases NTM infection (29.11%) among the 371 patients with branch enlargement. There was no statistical significance in cough, phlegm, hemoptysis and fever between the two groups (P>0.05). The proportion of chest tightness and shortness in NTM group was significantly higher than that in control group (P<0.05). Six mycobacterium species were isolated from 108 patients with NTM infection, including 57 intracellular, 22 abscess, 14 avium, 7 Kansas, 5 terrapin and 3 flavus. There were significant differences in age, sex, smoking history, place of residence, number of brandished lobes, hypoproteinemia, pulmonary cavities, and number of CD4+T lymphocytes between the two groups (P<0.05). Multivariate logistic regression analysis showed that smoking history >20 years (OR=1.692), number of branchial dilated lobe ≥5 (OR=2.671) and thin-walled cavity (OR=2.458) were independent risk factors for NTM infection in elderly patients with branchial dilated lobe (P<0.05). Conclusion Elderly patients with bronchiectasis in Huai 'an area have a high risk of NTM infection, which are mainly Mycobacterium intracellular and mycobacterium abscess. The main risk factors are the number of bronchiectasis lung ≥5, smoking history >20 years, and thin-walled cavity. Patients should actively quit smoking, improve the body immunity, and prevent NTM infection in patients with bronchiectasis.

The symbiotic bacterium exists in the intestines of entomopathogenic nematodes and is a potential biological agent.Systematic classification of these bacteria is scarce in China.In this paper,seven strains of symbiotic bacteria from local entomopathogenic nematodes were identified by both observation of mor-phology,physiological,biochemical characteristics and sequence analysis of 16S rDNA fragments.

OBJECTIVETo investigate the effect of homoharringtonine (HHT) on proliferation and apoptosis of CML cell line K562 cells and to explore its possible mechanism through mTOR pathway.

METHODSK562 cells were cultured with different concentrations of HHT or in its combination with mTOR inhibitor rapamycin (RAPA) for 24 hours. The cell viability was analyzed by CCK-8 assay, the cell apoptosis was detected by flow cytometry, the expressions of BCL-6, Caspase-3 and mTOR signal pathway related proteins was assayed by Western blot, the expression of BCL-6 mRNA was determined by RT-PCR.

RESULTSThe HHT inhibited proliferation and induced apoptosis of K562 cells in a concentration-dependent manner(r=0.970). With the increasing of HHT concentration, the expression level mTOR signal pathway related proteins increased(r=0.908), while the mRNA and protein expression levels of BCL-6 decreased(r=-0.961, r=-0.981), as compared with the HHT alone, the combination of HHT with RAPA could down-regulate the expression of mTOR signal pathway related protein and caspase-3, and up-regulated expression of BCL-6.

CONCLUSIONHHT induces apoptosis of K562 cells by inhibiting BCL-6 expression through mTOR signal pathway.