The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea，and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.

Objective To discuss the clinical value of 99Tcm-sestamibi(99Tcm-MIBI) myocardial perfusion imaging on detecting myocardial ischemia in children with Kawasaki disease(KD) at convalescence period.Methods Twenty-one children wih KD at convalescence period were divided into 2 groups according to results of echocardiography.Four cases with coronary artery dilation,17 cases without coronary artery dilation.All cases accepted dipyridamole stress myocardial perfusion imaging.These patients who had positive results were given rest myocardial perfusion imaging again next day.Results Among 21 cases,9 cases(42.8%) were positive in perfusion imaging.Four cases with coronary artery dilation showed myocardial ischemia in different degree detected by myocardial perfusion imaging.Among 17 cases without coronary artery dilation,5 cases(29.4%) were positive.Conclusions Compared to echocardiography,99Tcm-MIBI myocardial perfusion imaging can objectively evaluate the location,extent and degree of myocardial ischemia of children with KD.It will be a routine test in observing its phase development.

The primary object of this fundamental research was to survey the synergistic cardiovascular effects of iptakalim, a novel ATP-sensitive potassium channel (K(ATP)) opener, and clinical first-line antihypertensive drugs, such as calcium antagonists, thiazide diuretics and β receptor blockers by a 2 x 2 factorial-design experiment. It would provide a theoretical basis for the development of new combined antihypertensive therapy program after iptakalim is applied to the clinic. Amlodipine besylate, hydrochlorothiazide and propranolol were chosen as clinical first-line antihypertensive drugs. Blood pressure, heart rate (HR) and cardiac functions were observed in anesthetized normal rats by an eight-channel physiological recorder. The results showed that iptakalim monotherapy in a low dose could produce significant antihypertensive effect. There was no interaction between iptakalim and amlodipine on the maximal changes of systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial blood pressure (MABP), the left ventricular systolic pressure (LVSP), and the left ventricular end-diastolic pressure (LVEDP) (P > 0.05). However, the effects of combination iptakalim/amlodipine on the maximal changes of SBP, DBP, MABP, LVSP and LVEDP were more obvious than those of iptakalim or amlodipine monotherapy. And there was strong positive interaction between iptakalim and amlodipine on the maximal changes of HR (P>0.05). According to the maximal changes of DBP, MABP, LVSP and LVEDP (P < 0.05) of combination iptakalim with hydrochlorothiazide, there was strong positive interaction between them. But there was no interaction between iptakalim and hydrochlorothiazide on the maximal drop of SBP and HR (P > 0.05). According to the maximal drops of DBP, MABP of combination iptakalim with propranolol, there was strong positive interaction between them (P < 0.05). But there was no interaction between iptakalim and propranolol on the maximal changes of SBP, LVSP, LVEDP and HR (P > 0.05). In conclusion, it was the first time to study the effects of amlodipine, hydrochlorothiazide or propranolol, which had different mechanisms of action from iptakalim, on cardiovascular effects of iptakalim in anesthetized normal rats. This study proved that the combination of iptakalim with hydrochlorothiazide or propranolol respectively had significant synergism on lowering blood pressure, while the combination of iptakalim/amlodipine had additive action on lowering blood pressure. Meanwhile the antihypertensive effect was explicit, stable and long-lasting. Iptakalim thus appears suitable for the clinical treatment of hypertensive people who need two or more kinds of antihypertensive agents.
Amlodipine ; pharmacology ; Animals ; Antihypertensive Agents ; pharmacology ; Blood Pressure ; drug effects ; Drug Synergism ; Heart Rate ; Hydrochlorothiazide ; pharmacology ; Hypertension ; Propranolol ; pharmacology ; Propylamines ; pharmacology ; Rats

OBJECTIVETo study the effects of iptakalim (IPT) on pressure-overload induced cardiac remodeling in rats, and investigate correlation between this protection effects and plasma PGI2 content.

METHODThe pressure-overload induced cardiac remodeling model was induced by abdominal aorta constriction for 6 weeks, and the rats were divided into 5 groups repectively: (1) sham group, (2) control group, (3) IPT 3 mg/kg group (IPT 3), (4) indomethacin 2 mg/kg group (Indo 2), (5) indomethacin 2 mg/kg + IPT 3 mg/kg group (Indo 2 + IPT 3). RM6000 eight channel physiological recorder was used to record haemodynamics index, heart weight was weighed and the cardiac remodeling index was calculated, HE stain and Masson's stain were employed to perform histological analysis, colorimetric method was used to detect the hydroxyproline content in cardiac tissue, radioimmunological method was used to measure the plasma PGI2 content.

RESULTSAfter 42 days of aortic banding, the hyperdynamic circulation state, cardiac remodeling and decreased plasma PGI2 content were observed in the model group compared with those in the sham group, which were effectively reserved by treatment with IPT 3 mg/kg. Single-use indomethacin led to further deterioration of this pathophysiological changes, however, combination administration of IPT 3 mg/kg prevented these from worsening characteristic by ameliorating hyperdynamic circulation state and cardiac remodeling, augmnent plasma PGI2 content.

CONCLUSIONIPT can significantly reverse abdominal aorta binding/pressure-overload induced cardiac remodeling, its mechanism may contribute to binding K(ATP) channel in endothelial cells, ameliorating endothelium cells function, augmenting PGI2 synthesis and secretion.

Animals ; Aorta, Abdominal ; surgery ; Constriction ; Endothelium, Vascular ; metabolism ; physiology ; Epoprostenol ; blood ; Hypertension ; blood ; physiopathology ; KATP Channels ; drug effects ; Male ; Propylamines ; pharmacology ; Rats ; Ventricular Remodeling ; drug effects

Objective To investigate the screening and prevalence of tuberculosis among freshmen in different schools in Baoding City,and provide reference for tuberculosis control in schools.Methods Screening data of tu-berculosis and tuberculin test(PPD)of freshmen from 156 schools in different regions of Baoding City from Septem-ber 2021 to March 2022 were collected.PPD screening results of students from different regions and different school stages were analyzed and compared.Results A total of 68 177 freshmen from 156 schools were investigated for suspected symptoms and close contact history of pulmonary tuberculosis.PPD screening was conducted on 63 939 students.13 821 students were PPD positive,with a positive rate of 21.62％.3 083 students were strongly posi-tive,with a strong positive rate of 4.82％.15 cases of tuberculosis were found,and the reported incidence was 23.46/100 000.PPD positive rate and strong positive rate as well as incidence of tuberculosis in students in different school stages presented statistically significant differences(all P＜0.01).Positive rate and strong positive rate in students in different school stages showed upward trends(all P＜0.01).PPD positive rate and strong positive rate of students from schools in plain and mountainous areas presented statistically significant differences([22.28％vs 17.89％];[4.85％vs 3.62％],both P＜0.01).PPD positive rate and strong positive rate between students from boarding junior school and non-boarding junior school were significantly different,respectively([23.94％vs 21.60％];[5.07％vs 3.56％],both P＜0.01).Conclusion It is necessary to strengthen tuberculosis screening and health education for freshmen,especially those from boarding schools in plain areas,screening latent Mycobac-terium tuberculosis infection as early as possible,take corresponding measures to prevent and control the spread of tuberculosis,and reduce the risk of tuberculosis.

OBJECTIVE Lapachol is a natural naphthoquinone compound that possesses extensive biological activities. The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in vivo, as well as the potential mechanisms. METHODS The antitumor effect of lapachol was firstly evaluated in the C6 glioma model in Wistar rats. The effects of lapachol on C6 cell proliferation, apoptosis and DNA damage were detected by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS)/ phenazinemethosulfate (PMS) assay, hoechst 33358 staining, annexinⅤ-FITC/PI staining, and comet assay. Effects of lapachol on topoisomerase I (TOP I) and topoi?somerase Ⅱ (TOP Ⅱ) activities were detected by TOP Ⅰ and TOP Ⅱ mediated supercoiled pBR322 DNA relaxation assays and molecular docking. TOPⅠ and TOPⅡ expression levels in C6 cells were also determined. RESULTS High dose lapachol showed significant inhibitory effect on the C6 glioma in Wistar rats (P<0.05). It was showed that lapachol could inhibit proliferation, induce apoptosis and DNA damage of C6 cells in dose dependent manners. Lapachol could inhibit the activities of both TOPⅠ and Ⅱ. Lapachol-TOPⅠ showed relatively stronger interaction than that of lapachol-TOPⅡ in molecular docking study. Also, lapachol could inhibit TOPⅡ expression levels, but not TOPⅠ expression levels. CONCLUSION These results showed that lapachol could significantly inhibit C6 glioma both in vivo and in vitro, which might be related with inhibiting TOPⅠ and TOPⅡ activities, as well as TOPⅡ expression.

OBJECTIVETo investigate the work-related musculoskeletal disorders among automobile assembly workers, to discusses the related risk factors and their relationship.

METHODThe selected 1508 automobile assembly workers from a north car manufacturing company were regarded as the study object. The hazard zone jobs checklist, Nordic musculoskeletal symptom questionnaire (NMQ) and pain questionnaire were used to perform the epidemiological cross-sectional and retrospective survey and study for the General status, awkward ergonomics factors and related influencing factors, and musculoskeletal disorders of workers.

RESULTSThe predominant body sites of occurring WMSDs among automobile assembly workers were mainly low back, wrist, neck and shoulders, the predominant workshop section of occurring WMSDs were mostly concentrated in engine compartment, interior ornament, door cover, chassis and debugging section. The predominant body site of WMSDs among engine compartment and chassis section workers was low back, interior ornament workers were low back and wrist, door cover workers was wrist, chassis workers was low back, debugging workers were neck and low back. Neck musculoskeletal disorders had the trend with the increase of a body height; Smoking may increase the occurrence of musculoskeletal disorders.

CONCLUSIONThe WMSDs appears to be a serious ergonomic proble assem among automobile assembly workers, predominant occurring site of WMSDs is with different workshop section, its characteristics is quite obvious, probably related to its existing awkward work position or activities. The worker height and smoking habits may be important factors which affect musculoskeletal disorders happen.

Adult ; Cross-Sectional Studies ; Cumulative Trauma Disorders ; epidemiology ; Ergonomics ; Humans ; Male ; Musculoskeletal Diseases ; epidemiology ; Occupational Diseases ; epidemiology ; Prevalence ; Retrospective Studies ; Risk Factors ; Surveys and Questionnaires ; Young Adult

Objective To investigate the effect of emodin on spinal cord edema induced by acute spinal cord injury (SCI) and its mechanism. Methods A total of 180 healthy female Sprague-Dawley rats were randomly divided into sham group(group A),model group(group B),and low-dose group(group C),middle-dose group(group D)and high-dose group(group E)of emodin,with 36 cases in each group.The SCI model was established with the modified Allen's method.Function-al recovery was evaluated with Basso-Beattie-Bresnahan(BBB)score and inclined plate test three days,seven days,14 days and 28 days after modeling.Three days after modeling,the pathological changes of the spinal cord were observed by HE staining; the water content of spinal cord was detected by dry-wet weight method, the blood-spinal cord barrier(BSCB)permeability was detected by Evans blue(EB)staining,and the expression of aquaporin-4 (AQP-4) and matrix metalloproteinase-2 (MMP-2) mRNA and protein were detected by RT-PCR and Western blotting respectively.Results The BBB score and inclined plate scores were better in groups C,D and E than in group B(P<0.05)seven days, 14 days and 28 days after modeling, especially in group E (P<0.05). Three days after modeling, HE staining showed that there was a large hemorrhage in the section of group B,the nerve cells were swollen and damaged, and a large number of inflammatory cells were infiltrated,the tissue gap was widened and the edema was severe. The above pathological changes were better in groups C,D and E than in group B,especially in group E.The spi-nal cord water content was higher in group B than in group A(P<0.05),and was lower in groups D and E than in groups B and C(P<0.05).EB content was higher in group B than in group A(P<0.05),and was lower in groups C, D and E than in group B (P<0.05). The expression of AQP-4, MMP-2 mRNA and protein were lower in groups C,D and E than in group B(P<0.05),especially in group E(P<0.05). Conclusion Emodin can alleviate spinal cord edema,and improve hind limb movement function after SCI,which could be related with the down-regulation of AQP-4 and MMP-2 expression,and the reduction of the permeability of BSCB.

Objective To investigate the effect of cold-inducible RNA binding protein(CIRP)on the viability of cryopreserved sciat-ic nerve and nerve regeneration after allograft. Methods Sciatic nerve segments of 15 mm from male Sprague-Dawley rats were placed in DMEM solution and pretreat-ed with 4 ℃, 15 ℃ and 32 ℃ for 24 hours (group A, group B and group C, respectively). Fresh nerve group (group D)without pretreatment was set up.The mRNA and protein level of CIRP was detected by RT-PCR and Western blotting,respectively.The above nerves were cryopreserved in liquid nitrogen for four weeks.The via-ble cells of the nerve segments after cryopreservation were observed by calcein-AM/propidium iodide staining. The expression of Bax and Bcl-2 was detected by Western blotting.After cryopreservation,the nerve segments were cultured in vitro for one week, the protein level of nerve growth factor (NGF) and glial cell line-derived neurotrophic factor(GDNF)was detected by Western blotting.In addition,the above four cryopreserved groups were transplanted to the Wistar rats by allografting(groups A',B',C'and D').Fresh nerve allograft group(E')and isograft group(F')were set up.At four-week post operation,the expression of CD4of the nerve and plasma level of interleukin(IL)-6 and interferon(IFN)-γ were detected by immunohistochemistry and ELISA,respectively.At 20-week postoperation, the muscle compound action potential (CMAP) and motor nerve conduction velocity (MNCV) were examined by electrophysiological examination. The number and the thickness of myelinated nerve fibers were analyzed by toluidine blue staining. The ultrastructure of the sciatic nerve was observed by electron microscopy. Results The mRNA and protein of CIRP were significantly higher in group C than in groups A and B(P<0.05).After 4 weeks of cryopreservation,compared with groups A,B and D,the viable cells increased,the expression of Bax decreased and the expression of Bcl-2 increased in group C (P<0.05). The expression of NGF and GDNF in-creased in group C than in groups A and B(P<0.05).At four-week postoperation,the expression of CD4and plas-ma concentration of IL-6 and IFN-γ significantly decreased in group C'than in group E'(P<0.05),however,no significant difference was found in group C'compared with groups D'and F'(P>0.05).At 20-week postopera-tion, CMAP, MNCV, the number of axon, and thickness of myelin sheath were significantly better in group C' than in groups A',B',D'and E'(P<0.05).Compared with groups A',B',D'and E',the myelinated nerve fibers were more, the fiber thickness was more uniform, the fiber distribution was wider, and the myelin sheath was thicker in groups C'and F'. Conclusion CIRP was induced at 32℃in the sciatic nerve,which exerted a significant protective effect on the viability of the nerves during cryopreservation,and promoted nerve regeneration and functional recovery after transplanta-tion.

OBJECTIVETo investigate the protective effects of Shengui tablet (Chinese Traditional Medicine) on experimental cerebral ischemia by acute cerebral ischemia hypoxia in mice and bilateral ligation of the carotid artery in rats.

METHODSIn the acute cerebral ischemia hypoxia model, the mice were randomly divided into control group, low-, middle- and high-dose (0.16, 0.33 and 1.00 g/kg) groups of Shengui tablet, after oral treatment for 30 d, gasping time of isolated heads of mice were observed. In bilateral ligation of the carotid artery cerebral ischemia model, the rats were randomly divided into control group, model group and low-, middle-, high-dose (0.072, 0.149 and 0.450 g/kg) groups of Shengui tablet. After oral treatment for 7 d, the cerebral index, superoxide dismutase (SOD) activity and the content of malondialdehyde (MDA) were measured.

RESULTSCompared with the control model, Shengui tablet middle- and high-dose could significantly prolong gasping time of isolate heads of mice. Compared with model group, Shengui tablet low-, middle- and high-dose could significantly decrease the cerebral index and enhance SOD activity in brain tissue; only high-dose could reduce the content of MDA.

CONCLUSIONShengui tablet has significant protective effect on the cerebral ischemia.

Animals ; Brain ; metabolism ; Brain Ischemia ; metabolism ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; Female ; Male ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred Strains ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism