Abstract: Objective To express Mycobacterium tuberculosis Rv2626c protein in Escherichia coli (E. coli) and study the antigenicity of the purified recombinant Rv2626c protein. Methods The amino acid sequence of Rv2626c protein from Mycobacterium tuberculosis H37Rv strain (accession number: CCP45424.1) in GenBank was retrieved and converted into the corresponding DNA sequence according to the codon preference of E. coli. This DNA sequence was synthesized and cloned into pET24a(+) plasmid to construct pET24a(+)-Rv2626c recombinant plasmid. This plasmid was transformed into E. coli BL21(DE3) cells, and the expression of Rv2626c protein was induced under various conditions of isopropyl β-D-thiogalactopyranoside (IPTG) concentrations, temperature, and period. The recombinant Rv2626c protein was identified by SDS-PAGE and Western Blot. The recombinant Rv2626c protein was purified by nickel chelate affinity chromatography and used to immunize violet blue rabbits to prepare anti-Rv2626c anti-serum. The specificity and titer of the serum were respectively detected by Western Blot and enzyme-linked immunosorbent assay (ELISA). Results The recombinant plasmid pET24a(+)-Rv2626c was successfully constructed. SDS-PAGE analysis showed that recombinant Rv2626c was expressed in the recombinant plasmid transformed E. coli with IPTG induction, with a molecular weight of about 14 500, and the size was consistent with the expectation. The optimal expression condition for recombinant Rv2626c protein was at 31 ℃ with 1.0 mmol/L IPTG for 6 hours. The target protein was mainly present in a soluble form, which was consistent with the results of Western blot. The hyperimmunized serum with recombinant Rv2626c protein vaccination showed good specificity, with a titer of 1∶ 256 000 detected by ELISA. Conclusions Mycobacterium tuberculosis Rv2626c protein is successfully expressed in E. coli, and the purified protein has good purity and antigenic activity, laying the foundation for further reveals of its biological functions.

Objective: To analyze the association between different sleep behaviors and overweight and obesity of junior high school students in Yangzhou City, and to provide a basis for policies and interventions related to adolescent health management. Methods: A total of 1 589 students in grades 7-9 from two middle schools in Yangzhou City were selected using the cluster sampling method and were administered with sleep time, bedtime, social jetlag difference, and sleep habits. Results: Totally 64.38% were sleep deprived during the school days, 86.78% went to bed too late, 46.51% had a social jetlag of ≥1 h, and 37.44% took a nap every day(Incluldes holidays and school days). Social jetlag length was statistically different between grades( F =6.97, P < 0.01 ). Girls[(0.95±0.65)h] shown significantly higher social jetlag than the boys[(0.76±0.59)h]( t=6.19, P <0.01). Later bedtime on weekends, later wake up time on weekends and poor sleep behavior were risk factors for overweight and obesity in junior high school students( OR=1.20, 1.14, 1.04, P <0.05). Conclusion Junior high school students had less sleep and later bedtimes with the increase of grade, and weekend bedtimes,wake up times and poor sleep behavior were independently associated with the risk of overweight and obesity in junior high school students. Parents and schools should be instructed to pay attention to their sleep health and carry out adolescent sleep health guidance.

Lipophagy, the selective engulfment of lipid droplets (LDs) by autophagosomes for lysosomal degradation, is critical to lipid and energy homeostasis. Here we show that the lipid transfer protein ORP8 is located on LDs and mediates the encapsulation of LDs by autophagosomal membranes. This function of ORP8 is independent of its lipid transporter activity and is achieved through direct interaction with phagophore-anchored LC3/GABARAPs. Upon lipophagy induction, ORP8 has increased localization on LDs and is phosphorylated by AMPK, thereby enhancing its affinity for LC3/GABARAPs. Deletion of ORP8 or interruption of ORP8-LC3/GABARAP interaction results in accumulation of LDs and increased intracellular triglyceride. Overexpression of ORP8 alleviates LD and triglyceride deposition in the liver of ob/ob mice, and Osbpl8-/- mice exhibit liver lipid clearance defects. Our results suggest that ORP8 is a lipophagy receptor that plays a key role in cellular lipid metabolism.
Animals ; Mice ; Lipid Droplets ; Autophagy ; Autophagosomes ; Homeostasis ; Triglycerides

OBJECTIVES: Cardiac hypertrophy and fibrosis are major pathological manifestations observed in left ventricular remodeling induced by angiotensin II (AngII). Low-intensity pulsed ultrasound (LIPUS) has been reported to ameliorate cardiac dysfunction and myocardial fibrosis in myocardial infarction (MI) through mechano-transduction and its downstream pathways. In this study, we aimed to investigate whether LIPUS could exert a protective effect by ameliorating AngII-induced cardiac hypertrophy and fibrosis and if so, to further elucidate the underlying molecular mechanisms. METHODS: We used AngII to mimic animal and cell culture models of cardiac hypertrophy and fibrosis. LIPUS irradiation was applied in vivo for 20 min every 2 d from one week before mini-pump implantation to four weeks after mini-pump implantation, and in vitro for 20 min on each of two occasions 6 h apart. Cardiac hypertrophy and fibrosis levels were then evaluated by echocardiographic, histopathological, and molecular biological methods. RESULTS: Our results showed that LIPUS could ameliorate left ventricular remodeling in vivo and cardiac fibrosis in vitro by reducing AngII-induced release of inflammatory cytokines, but the protective effects on cardiac hypertrophy were limited in vitro. Given that LIPUS increased the expression of caveolin-1 in response to mechanical stimulation, we inhibited caveolin-1 activity with pyrazolopyrimidine 2 (pp2) in vivo and in vitro. LIPUS-induced downregulation of inflammation was reversed and the anti-fibrotic effects of LIPUS were absent. CONCLUSIONS These results indicated that LIPUS could ameliorate AngII-induced cardiac fibrosis by alleviating inflammation via a caveolin-1-dependent pathway, providing new insights for the development of novel therapeutic apparatus in clinical practice.

Abdominoscrotal hydrocele is rare in clinic. In the past, routine examination and diagnosis were difficult, easy to be misdiagnosed. The daily operations were mostly completed through the groin area or abdominal incision, the wound is large. The application of laparoscopy can clearly diagnose the abdominoscrotal hydrocele through "springing back ball" sign, and can cure the disease by laparoscopic resection of interperitoneal mass and closure of the internal ring. It is worthy of clinical application. In this article, we summarized and analyzed the clinical experience of 15 cases of children with abdominoscrotal hydrocele diagnosed and treated by laparoscopy, to explore the value of the laparoscopic technology in the diagnosis and treatment of the abdominoscrotal hydrocele.

Objective To investigate the application of venous superdrainage technique based anterolateral thigh flap based on oblique branch for the soft tissue defect of proximal shank.Methods Between October,2012 and March,2017,11 cases of the soft tissue defect in proximal shank were treated.There were 8 males and 3 females with a mean age of 43 (range,26-59) years.Causes of injury:7 cases of traffic injury,3 cases of bruise,1 case of machine wound;The defect sizes ranged from 10 cm×5 cm to 13 cm×10 cm,with fracture or plate exposure.Accord ing to the size of the defect,the distally based anterolateral thigh flap was designed to repair the defect.The flap sizes ranged from 14 cm×6 cm to 16 cm×8 cm.The oblique branch and their accompanying vein with great saphenous vein was anastomosed to construct the venous superdrainage,and the donor site suture directly.Three cases followed-up by WeChat,and 8 cases by return visit.Results Distally based anterolateral thigh flaps were derived from oblique branches,and no arteriovenous crisis occurred postoperatively.Eleven flaps healed well after operation.There were no distal end blood supply disorder.Followed-up for 6-12 months,the flaps survived well.The texture was closed to the surrounding skin,and no ulcer exposed.There were no serious complications in donor site.And the thighs were linear scars.Conclusion The distally based anterolateral thigh flap based on oblique branch have a long vascular pedicle.The venous superdrainage technique can effectively avoid venous crisis,and improve the survival quality of the flap and the survival rate.

Objective To retrospectively analyze the injury characteristics of victims and treatment strategies in the explosion accident on the 17th May 2018 in Xixia county (Xixia "May 17th" explosion accident). Methods Based on the practice featured in pre-hospital emergency of Henan province and Nanyang city Emergency Center in the explosion accident, a retrospective analysis for the Level Three medical rescue was conducted, where a total of thirteen survived victims in Xixia "May 17" explosion accident were studied retrospectively. The data included the gender, age, burned extent and depth of the patients, burns complicated by trauma, complication of burn, respiratory function maintenance, resuscitation during shock stage, skin grafting with excision and scab. Furthermore, the data of organ function and the effect of the 90-day comprehensive treatment for the burned victims wereanalyzed. Results completion the Level Three treatment on time, which was depended on the leading role played by the regional trauma centers was the main rescuing mode of the work in Xixia county, where the primary and secondary treatments were the key parts. The three-level treatment model includes: the local hospital acts as a level-one emergency medical institution, county hospitals function as secondary emergency medical institutions, and other higher medical institutions are the tertiary first aid medical institutions. The pre-hospital and in-hospital emergency procedures were initiated immediately after the large-scale explosive burn being identified, the key to the successfully rescue was to set up a comprehensive treatment team for burns and trauma. Rescue team should involve burn department and other related departments, including the departments of emergency, general surgery, orthopedic, thoracic surgery, neurosurgery, plastic surgery, intensive care unit, blood transfusion unit, anesthesiology, and interventional radiology, etc. All the thirteen burned patients were male, with inhalation injury, blast injury, hemopneumothorax, brain injury, bone fractures, and etc. Eight of them (61.54%) had multiple organ dysfunction syndrome (MODS). MODS mainly involved respiratory, circulatory, liver, gastrointestinal tract, kidney and coagulation function. With the multi-discipline treatment, the wound of 6 severely-burned patients started healing and can be discharged after keeping the patency of airway, applying resuscitation fluid and comprehensive treatments such as debridement and dressing change. Among 7 patients with extensive deep burns, one case with skull-based fracture, open craniocerebral, extensive intracranial hemorrhage and hemopneumothorax, died 9 hours later. Another case died within 24 hours after injury due to obvious exudation on the site of early incision and relaxation of wound. The escharotomy, micro-dermis and allograft skin transplantation were carried out for five cases with extensive deep burns from the 4th day after the recovery of shock. One week later, the second stage of microsphere skin transplantation was performed. But all died of sepsis or fungal infection. Conclusions MODS and infection often occur during the course especially for patients with extensive and deep burns due to the great explosion in Xixia county, most of whom were accompanied with MODS and infection. Therefore, assembling multi-discipline team for treating the group of explosively-burned patients can increase the survival rate and reduce the possibility of disability.

Objective To study the effect of cyclosporine combined with eleven acid testosterone on serum T helper cells in patients with chronic aplastic anemia.Methods A total of 86 patients with chronic aplastic anemia received treatment in our hospital were collected as the research objects,and divided into experimental group and control group,patients in the experimental group received eleven acid testosterone combined with cyclosporine therapy,patients in the control group received eleven acid testosterone therapy.Blood routine indexes,Th1,Th2,Th17 and Treg cells contents of the two groups before and after treatment were compared.Results Before treatment,hemoglobin content,leukocyte count,platelet count,peripheral blood Th1,Th2,Th17 and Treg cells contents of the two groups had no significant difference(P>0.05).After two courses of treatment,hemoglobin content,leukocyte count,platelet count of the experimental group were significantly higher than those of the control group.After two courses and four courses of treatment,peripheral blood Th1 and Th17 contents of experimental group were significantly lower than those of the control group,Th2 and Treg contents were significantly higher than those of the control group.Conclusion Eleven acid testosterone combined with cyclosporine treatment could be more effective to stimulate hematopoiesis,regulate immune function and T lymphocyte subsets content,it′s an effective method for the treatment of chronic aplastic anemia.

Objective To analyze and compare the epidemiological features of Monteggia fractures between the east and west areas in China from 2010 to 2011.Methods The data of Monteggia fractures treated from January 2010 through December 2011 in 63 hospitals in the east and west China were collected through the PACS system and case reports checking system.The data from the 35 east hospitals were classified as group A while those from the 28 west hospitals as group B.The analytic items included general situation,gender,age,and Bado classification.Results A total of 593 Monteggia fractures were included,including 428 males and 165 females,with a male to female ratio of 2.59:1.The constituent ratios of Bado classification,from high to low,were type Ⅰ (42.50％),type Ⅲ (33.22％),type Ⅱ (14.00％) and type ⅣV (10.29％).There were 376 patients in group A including 263 (69.95％) males and 113 (30.05％)females while 217 patients in group B including 165 (76.04％) males and 52 (23.96％) females.The median age in group B was 31 yearn (from 2 to 75 years),significantly older than that in group A [18 years (from 1 to 83 years)] (Z =2.877,P =0.000).The differences in constituent ratios of age and Bado classification were statistically significant between the 2 groups (P ＜ 0.05).The Monteggia fractures predominated in the age range of 0 to 10 years in group A while in the age range of 31 to 40 years in group B.The high risk fracture type was Bado type Ⅰ in group A and Bado type Ⅲ in group B.The Monteggia fractures accounted for 1.15％ of the forearm fractures and 0.19％ of the systemic fractures.Conclusions More males suffered from Monteggia fracture than females.In the east China,the high risk age range was from 0 to 10 years and the high risk type Bado type Ⅰ;in the west China,the high risk age range was from 31 to 40 years and the high risk type Bado type Ⅲ.The proportion of Bado type Ⅳ in the west China was significantly higher than in the east China.

Objective To measure the concentration of serum transthyretin (TTR) of patients with different stages of diabetic retinopathy (DR).Methods A total of 176 patients with diabetes mellitus were included in this study.There were 104 males and 72 females.The patients aged from 21 to 74 years,with the mean age of(56± 11) years.The diabetes duration raged from 1 to 30 years,with the mean diabetes duration of (10 ± 7) years.The HbA 1C was 5.2％-14.1％,with the mean HbA 1C of (8.6 ± 2.0)％.According to the fundus examination,58 patients had DR (33.0％),but the other 118 patients not (67.0％).For these DR patients,10 patients were in stage Ⅰ (5.7％),26 patients in stage Ⅱ (14.8％),8 patients in stage Ⅲ (4.5％),and 14 patients in stage Ⅳ (8.0％).The concentration of serum TTR was measured by enzyme-linked immunosorbentassay kit.The differences in the concentration of serum TTR between different DR stages were compared.Bivariate analysis was used to analyze the influencing factors of TTR.Results The concentrations of serum TTR of the patients without DR or with DR of stage Ⅰ to Ⅳ were (224.96±65.47),(383.68± 102.99),(247.44±63.21),(228.2 ± 45.89),(189.34± 70.12) mg/L,respectively.The difference between different DR stages was statistically significant (F=14.690,P＜ 0.001).Bivariate analysis showed that the concentration of TTR was correlation to DR (r=0.179,P=0.017).There was no correlation between the concentration of TTR and diabetes duration (r=-0.027,P=0.727),hypertension (r=0.018,P=0.810),hyperlipoidemia (r=0.101,P=0.182),and the use of insulin (r=-0.032,P=0.675).Conclusion The concentration of serum TTR was increased in early DR patients,and gradually decreased with the progression of DR.The concentration of TTR is correlated to DR.