Objective To clone and express the ns5a gene of hepatitis C virus(HCV) 1b strain DY.Methods By using the prokaryotic cell gene engineering,HCV ns5a gene was amplified with nested PCR from the plasmid HCV17 of HCV 1b strain DY full-length gene and inserted into the cloning pMD18-T vector.The cloned HCV ns5a gene was separated and subcloned into expression vector pET-28a and induced by IPTG in E.coli.BL21.The expressed product was identified by SDS-PAGE and Western-blot methods.Results Recombinant expression plasmid pet-28a-ns5a was constructed and expressed successfully.Conclusion HCV ns5a gene was cloned and expressed.This might be helpful for further studies on the nature and biological properties of the ns5a gene.

Objective： The current study was designed to clarify whether hepatopoietin (HPO) stimulates autonomous growth of hepatoma cell by autocrine loop. Methods： The authors conducted experiments in vitro with hepatoma cell lines. RT-PCR, ELISA and Western blot were used to examine HPO expression in hepatoma cells. Blocking effect of HPO by HPO neutralizing antibody was utilized and the changes of cell proliferation was observed. Results： HPO was expressed by hepatoma cells and secreted into the medium. Moreover, the HPO antibody inhibited specifically the autonomous proliferation of hepatoma cell and antagonized the stimulatory effect of concentrated conditioned medium derived from hepatoma cell HepG2. Conclusion： The results strongly suggest that HPO acts as an autocrine factor to maintain the autonomous growth of hepatoma cells.

Objective To understand the dietary nutrients among rural stranded children.Methods 2551 children aged 2 to 7, including 1278 stranded children in the rural areas and another 1273 children served as controls were selected, using multistage stratified cluster random sampling. Dietary survey was performed with three-day weighing dietary method and questionnaire on food intake. Data on diet were analyzed and evaluated by the Dietary Reference Intakes (DRIs) recommend by Chinese Nutrition Society, to evaluate the levels on energy and nutrient intake among stranded children in the rural areas. Results The dietary pattern among rural stranded children mainly consisted of grains and vegetables, but the intakes of animal products, fruits, and snacks were significantly less than in the control group. The intakes of three major energy-producing nutrients were below the recommend nutrient intake. Minerals as calcium, zinc, selenium, kalium and vitamins as vitamin A, B1, B2were insufficient.Most of the rural stranded children took nutrients insufficiently, with 50% lack of adequate energy and 80% of protein, 90% of minerals (calcium, zinc etc.) and vitamins (vitamin B1 and vitamin B2 etc.).Sources from high quality protein was insufficient, only consisting 35% of the total protein, but overabundant (over 64 % ) from the plants. The intake of plant-sourced iron was overabundant, accounted for 87%. Conclusion The dietary pattern was unsatisfactory with insufficient intake of energy-sourced proteins and some nutrients. The sources of energy, protein, and iron were mostly obtained from underbalaneed foods. It is necessary to improve the dietary nutrients status among rural stranded children aged 2-7 years.

Objective To explore knowledge, attitude, practice (KAP) and related determinants on nutrition among caregivers of those rural stranded children under 7 years of age in China and to provide evidence for setting up relevant health education program. Methods 1691 caregivers of the stranded children randomly selected were surveyed by a standard questionnaire. Logistic regression models were used to screen the determinants on KAP regarding nutrition. Results Rates on awareness, positive attitude and approprite behavior were lower in caregivers of children whose parents both left (47.8%, 55.4%, 41.8%, respectively) the countryside, when compared with those only one parent was away from home (59.9%, 59.5%, 38.0%, respectively). Data from multivariable logistic regression models showed that caregivers' KAP on nutrition was related to age, educational background, average family income, and willingness on the job as well as the age of the child. Conclusion Improving caregivers' KAP on nutrition and setting up appropriate health education program were in urgent need.

king condition before pregnancy were associated with the development of polydactyly and syndactyly.

OBJECTIVETo study the expression and location of early growth response gene-1 (Egr-1), transforming growth factor-beta(1) (TGF-beta(1)), fibronectin (FN) in silicotic rat and to discuss the role of Egr-1 in the development of silicosis.

METHODSSilicotic animal model of rat was established, and the expressions of Egr-1, TGF-beta(1), FN in various lung cells of silicotic rat were analysed by using immunohistochemical technique (SP) and the image analysis.

RESULTSThe expressions of Egr-1 in bronchial epithelial cell, pulmonary macrophage, alveolar epithelium cell and interstitial cell in lung silicotic tissue (gray values: 118.58 +/- 5.65 - 168.52 +/- 5.67) were higher than those of controls (gray values: 166.23 +/- 5.23 - 188.12 +/- 8.35) during 1 - 28 days, and the expression was mainly in nucleus; the expressions of TGF-beta(1) in these cells (gray values: 123.49 +/- 5.65 - 170.24 +/- 3.56) were also higher than those of controls (166.53 +/- 6.25 - 198.56 +/- 4.53), and the expression was mainly in cytoplasm. The expressions of FN in bronchial epithelial cell, pulmonary macrophage and alveolar epithelial cell (gray values: 150.32 +/- 6.54 - 201.54 +/- 7.38) were lower, while those in interstitial cell (gray values: 121.43 +/- 5.65 - 167.55 +/- 6.35) were higher than those of controls. The changes of TGF-beta(1) and Egr-1 expression level in bronchial epithelial cell, pulmonary macrophage, alveolar epithelium cell and interstitial cell were synchronous during the experiment (1 - 28 days). Both of them were correlated with each other (r = 0.61, P < 0.01), while the expression of FN was not correlated with Egr-1, but correlated to TGF-beta(1) in interstitial cell (r = 0.46, P < 0.01).

CONCLUSIONSilicon dioxide could up-regulate the expression of nuclear transcription factor Egr-1 in several kinds of cell in lung. The activated Egr-1 may coordinate the expression of TGF-beta(1) and FN to regulate the development of silicosis.

Animals ; DNA-Binding Proteins ; analysis ; physiology ; Disease Models, Animal ; Early Growth Response Protein 1 ; Fibronectins ; analysis ; physiology ; Immediate-Early Proteins ; analysis ; physiology ; Immunohistochemistry ; Lung ; chemistry ; physiopathology ; Rats ; Silicosis ; etiology ; metabolism ; Transcription Factors ; analysis ; physiology ; Transforming Growth Factor beta ; analysis ; physiology

Objective To investigate the treatment and prevention of infection after alveolar crest onlay bone graft. Methods From January 2006 to May 2010, 11 infection cases after onlay graft on alveolar crest were reviewed to evaluate the infection time, clinical situation, treatment measure, and therapeutic effect Results The infection of all 11 cases occurred about 15 days after bone graft, which showed either soft tissue fistulae or bone graft exposure in the oral cavity. Three cases failed because of persistent infection.The infection of the other 8 cases was controlled after a series of comprehensive therapy, and most of the bone graft was reserved and implant restoration finally completed. Conclusions After the effective and comprehensive therapy, infected bone graft can be reserved. But to ensure the survival rate of bone graft, the most important thing is to prevent infection in perioperative period.

Objective:To explore the clinical application value of immediate breast reconstruction with implants after nipple-areola complex-sparing subcutaneous mastectomy for nonpuerperal mastitis .Methods:This study observed 22 women with nonpuerperal mastitis undergoing immediate breast reconstruction with implants after nipple-areola complex-sparing subcutaneous mastectomy from June 2012 to September 2014 .The perioperative results and follow-up records were analyzed . Results:The operation time was 104-132 min (mean 118 min) ,blood loss was 87-119 mL (mean 94 mL ) ,and the time of drainage tube removal was 10-21 d after operation .The length of hospitalization was 10-14 d (mean 12 d) ,and the average expenditure was about 20 000 RMB .There was no wound subcutaneous effusion ,no skin flap necrosis and no ischemia or necrosis in nipple-areola complex .In the evaluation of postoperative breast appearance ,9 cases were excellent ,13 cases were good ,and the excellent and good rate was 100% .All patients were followed up for 4-36 months (median 23 months) after operation .During the follow-up period ,no local recurrence ,breast deformation or fiber cystis contracture were formed .No upper extremity edema or dysfunction were found .Conclusions:Immediate breast reconstruction with implants after nipple-areola complex-sparing subcutaneous mastectomy possesses the advantage of minimal invasion , safety , quick postoperative recovery and simple operation for patients with nonpuerperal mastitis and the appearance of reconstructed breast is optimal .

Objective To design a self-aid pressurized heating tourniquet to decrease the mortality and morbidity of post-traumatic hemorrhage of limbs in highland cold regions.Methods The tourniquet was in the form of long strip and consisted of a gas generating heating mechanism,an inflatable cuff,a decompression mechanism and a micro timer.The gas generating heating mechanism was composed of an inner cylinder and an outer cylinder,which were fixed with screws;the inflatable cuff had two variations for the upper limb and lower limb respectively,with fabric for fixing on the outer side and an airbag set on the inner side;the decompression mechanism had a pressurized valve as its main component;the electronic micro timer was secured to the slot on the gas generating heating mechanism by an insertion clasp.Results The tourniquet developed enabled self-aid hemostasis in 1 to 2 s for post-traumatic hemorrhage of limbs in highland cold regions,and could effectively prevent the affected limb from losing temperature.Conclusion The tourniquet developed gains advantages in easy operation,hemostasis,heat preservation and adjustable pressure,which is of significance for enhancing the self-aid ability for post-traumatic hemorrhage of limbs in highland cold regions.[Chinese Medical Equipment Journal,2024,45(3):111-114]

This study aims to explore the effect and mechanism of Jiao-tai-wan (JTW) on systemic and tissue-specific inflammation and insulin resistance in obesity-resistant (OR)rats with chronic partial sleep deprivation (PSD).OR rats with PSD were orally given JTW and Estazolam for 4 weeks.The amount of food intake and metabolic parameters such as body weight increase rate,fasting plasma glucose (FPG),fasting insulin (FINS),homeostasis model assessment-insulin resistance (HOMA-IR) and plasma inflammatory markers were measured.The expression levels of circadian proteins cryptochrome 1 (Cry1)and cryptochrome 2 (Cry2) in hypothalamus,adipose and liver tissues were also determined.Meanwhile,the mRNA expression of inflammatory markers,activity of nuclear factor kappa B (NF-κB) p65 protein,as well as the expression levels of insulin signaling pathway proteins in hypothalamus,adipose and liver tissues were measured.Additionally,cyclic adenosine 3',5'-monophosphate (cAMP) and activity of vasodilator-stimulated phosphoprotein (VASP)in hypothalamus tissue were measured.JTW significantly decreased the body weight increase rate and food intake,ameliorated systemic inflammation and insulin resistance.JTW effectively ameliorated inflammation and increased PI3K/AKT signaling activation in hypothalamus,adipose and liver.Interestingly,all these changes were associated with the up-regulation of circadian gene Cryl and Cry2 protein expression.We also found that in hypothalamus tissue of P SD rats,down-regulation of Cry 1 and Cry2 activated cAMP/PKA signaling and then led to inflammation,while JTW inhibited this signaling.These results suggested that JTW has the beneficial effect on ameliorating inflammation and insulin resistance in partially sleep-deprived rats by up-regulating Cry expression.