The aim of this study is to establish stable transfected cell lines which could produce SPAG4L protein labeled with Myc and His tags in vitro. The open reading frame (ORF) of human SPAG4L was amplified by PCR and the fragments were cloned into eukaryotic expression vector pcDNA3.1/myc-His(-)A. The recombined plasmids of pcDNA3.1/myc-His(-)A/SPAG4L were verified by sequencing and digestion with enzymes. Then, the recombined plasmids were introduced into HeLa cells and screened by G418. Western blotting was performed to detect the expression of SPAG4L and its tags in stable transfected cell lines. SPAG4L and its tags were expressed in the stable cell lines transfected with pcDNA3.1/myc-His(-)A/SPAG4L, but not in the control group. Further study showed that SPAG4L colocalized with the endoplasmic reticulum(ER) marker PDI by immunofluorescence. The stable transfected cell lines established in this study will provide a powerful tool for further studies such as co-immunoprecipitation and pull-down.
Carrier Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Female ; Genetic Vectors ; genetics ; HeLa Cells ; Histidine ; genetics ; Humans ; Male ; Proto-Oncogene Proteins c-myc ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Transfection

Objective To investigate the impact of glucose metabolism-related protein 1 (GMRP1) on the regulation of c-Myc gene transcription, to establish GMRP1 gene knockout mouse model, and to study the influence of GMRP1 on glucose metabolism. Methods Chromatin immunoprecipitation-PCR (ChIP-PCR) was utilized to screen out the genes transcriptionally regulated by GMRP1. Luciferase reporter system was applied to assay the regulation of c-Myc promoter activity by GMRP1. GMRP1 knockout mice were constructed and validated by PCR and western blotting. Body weight and random blood glucose were measured in both knockout and wild type mice fed with either chow diet or high-fat diet. The levels of glucose metabolism in mice of 20 or 28 weeks old were evaluated by intraperitoneal glucose tolerance test. Results GMRP1 was capable of binding to the promoter of c-Myc gene and activating the expression of c-Myc gene. There were no significant differences in body weight, random blood glucose or glucose tolerance between GMRP1-deficient and wild type mice fed with either chow diet or high-fat diet (all P > 0.05). Conclusion GMRP1 may activate the transcription of c-Myc gene. GMRP1 deficiency exerted no significant effects on mouse glucose metabolism.

OBJECTIVETo probe into the mechanism and the best program of acupuncture for slimming.

METHODSThe patients were randomly divided into a control group (n = 60) treated with diet control and exercise, and a treatment group treated with acupuncture, auricular point sticking, TDP radiation. Changes of body weight, cholesterol (TC), triglyceride (TG) and blood sugar (BS) before and after treatment were investigated.

RESULTSThe effective rate of the treatment group (91.6%) was better than that of the control group (28.3%).

CONCLUSIONAcupuncture, auricular point sticking and TDP radiation combined with diet control and exercise are a better program for slimming, and decreasing appetite, promoting lipid decomposition metabolism are the mechanism of acupuncture in slimming.

Objective To explore the clinical characteristics, treatment and prognosis of bronchopulmonary dysplasia (BPD) in extremely preterm infants. Method The clinical data of extremely preterm infants with gestational age ＜ 28 weeks hospitalized between August 2014 and August 2017 were retrospectively analyzed. According to oxygen inhalation and BPD clinical grading at 28 days after the birth, extremely preterm infants were divided into non-BPD group, mild, moderate and severe BPD groups, and their clinical characteristics, treatment and prognosis were discussed. Results A total of 205 extremely preterm infants were enrolled, with an average gestational age of (26.2±1.2) weeks and an average birth weight of (0.87±0.19) kg. Among them, 46 infants were gave up treatment, 15 died, and 144 (70.3％) were discharged eventually. There were statistically significant differences in the rates of patent ductus arteriosus (PDA) and infection among groups with different degrees of BPD (P＜0.01) . The incidence of PDA was higher in the moderate and severe BPD groups, and the infection rate was higher in the severe BPD group. The oxygen use time, length of stay, hospitalization cost, glucocorticoid treatment rate and incidence of pulmonary sequelae among groups with different degrees of BPD were significantly different (P＜0.01) . With oxygen use time, length of hospital stay and hospitalization cost increased gradually over the aggravation of BPD. Glucocorticoids were used in all moderate and severe children, and the incidence of pulmonary sequelae was higher. Conclusion BPD still is an important complication affecting the prognosis of extremely preterm infants. Early prevention and systematic treatment can alleviate severe sequela.

Immune checkpoint therapy is a new mode of tumor therapy. At present, many kinds of PD-1/PD-L1 inhibitors have been approved for clinical application on various tumors, such as malignant melanoma, lung cancer, bladder cancer, lymphoma, etc. However, with the gradual promotion and application in clinical practice, various immune-related adverse reactions caused by PD-1/PD-L1 inhibitors have attracted extensive attention. Especially in recent years, a number of adverse reactions related to endocrine gland injury have been reported. This paper reviews the research status and corresponding clinical treatment methods of endocrine gland related adverse reactions caused by PD-1/PD-L1 inhibitors.

Objective:To investigate the demand and influencing factors of home care social support services for the elderly with dementia in urban areas, and provide a decision-making basis for improving home care social support services for the elderly with dementia.Methods:This was a cross-sectional study. From March 2021 to March 2022, 386 family caregivers of elderly people with dementia were randomly selected from tertiary hospitals in four central urban areas of Wuhan by stratified sampling method. A questionnaire was used to investigate the current situation and attributes of the demand for home care social support services for the elderly with dementia in urban areas. The importance of the demand for home care social support services for the elderly with dementia in urban areas was calculated by the Better-Worse coefficient method and sensitivity formula method. The optimal scale regression model was used to analyze the influencing factors and influencing forces of independent variables on service demand.Results:The score of social support service demand for home care was (88.6 ± 29.71) points. The Better coefficient of demand for all social support services was greater than the absolute value of the Worse coefficient. One requirement (pressure ulcer prevention and treatment guidance) was a one-dimensional attribute, and the remaining 29 items were charm attributes. The top 6 items in importance ranking were 2 items of informational support (pressure ulcer prevention and treatment guidance; telephone counselling guidance for family caregivers), 1 items of instrumental support (financial assistance for family caregivers) and 3 items of social companionship support(cultural recreation/physical activity activities, door-to-door accompaniment, accompany the elderly to social activities). Optimal scale regression model test: all four models of social support service demand were statistically significant ( F values were 2.75-4.23, all P <0.01). The gender, education level, number of children of the elderly with dementia, age,education level, self rated physical condition of family caregivers, the average monthly income of families, and the relationship between family caregivers and the elderly with dementia were the influencing factors of self-esteem support (all P<0.05). The educational level, marital status, type of dementia of the elderly with dementia, educational level, self rated physical condition of family caregivers, the average monthly income of the family, and the relationship between family caregivers and the elderly with dementia were the influencing factors of informational support (all P <0.05). The educational level and type of dementia of the elderly with dementia, the educational level, self rated physical condition of family caregivers, and the average monthly income of families were the influencing factors of social companionship support (all P <0.05). The type of dementia of the elderly with dementia, the self rated physical condition of family caregivers, the average monthly income of families, and the relationship between family caregivers and elderly relatives were the influencing factors of instrumental support (all P <0.05). Five times for caregiver human resources, 3 times for family economic resources and 3 times for the elderly with dementia, and 1 time for relatives resource factors were the top 3 times of independent variable influence force entered the ranking. Conclusions:The demand for social support services for home care of the elderly with dementia is low, with the characteristics of expectant tendency and centralized demand. The provision of home-based care social support services should give priority to meeting the needs of "pressure ulcer prevention and treatment guidance", and provide financial assistance, counseling and guidance, social companionship and other services as much as possible. Caregiver human resources are the key factors affecting home care social support services, family economic resources and the elderly with dementia are important factors, it is recommended to fully tap the potential of relatives resources and community resources.

In order to improve the quality and yield of Gastrodia elata f. glauca,determine the suitable Armillaria strains for the accompanying experiment in Xiaocaoba,Yiliang,four Armillaria strains were selected. They were used for G. elata cultivation,and the gene sequence,r DNA-ITS,β-tubulin and EF1-α of four Armillaria strains,were compared and analyzed. The yield was mesured in November which was based on previous laboratory research. The tubers were washed and steamed,then dried and powdered. The content of gastrodin and p-hydroxybenzyl alcohol was determined by UPLC,the polysaccharide was determined by phenol-concentrated sulfuric acid method. The results showed that the strains M1,M2,M3 and M4 were Armillaria gallica group but there were differences in the yield and active ingredient content when they were cultivated with the same G. elata. The yield of G. elata( Jian Ma) was the lowest when cultivated with Armillaria strain M3,but it was not the same when used M1,0. 981 kg·m-2,the highest yield in the four stains.The content of gastrodin was 0. 581%,the total content of gastrodin and p-hydroxybenzyl alcohol was 0. 595%,when accompanied with M1 strains. It was higher than other strains. The content of G. elata polysaccharide was 2. 132%,which was similar to the content of M3 strain,higher than that of M2 and M4 strain. Selecting phylogenesis of Armillaria strians,the content of active ingredient,and the yield as indicators,it was concluded concerned that the M1 strain was the best of four strains. The results will provide a theoretical basis and guidance for higher yield and quality in cultivation of G. elata in Yiliang.
Armillaria/physiology* ; Gastrodia/microbiology* ; Phylogeny ; Plant Tubers/chemistry* ; Plants, Medicinal/microbiology*

Objective To analyse the risk factors associated with spontaneous intestinal perforation (SIP) in extremely premature infants/extremely low birth weight infants. Method From January 2015 to December 2018, infants with gestational age (GA)<28 weeks or birth weight (BW)<1000 g admitted to our neonatal intensive care unit were enrolled to the retrospective nested case-control study.The clinical data of SIP infants (SIP group) and infants with the same GA but without SIP (control group) were randomly selected and compared. Multivariable Logistic regression was used to analyse the risk factors of SIP. Result A total of 409 extremely premature infants/extremely low birth weight infants were born during the study period. Among them, 25 SIP infants and 55 controls were enrolled. The incidence of SIP in infants with GA 22～25 weeks was 11.8％(16/136), which is higher than infants with GA 26～27 weeks (2.0％, 5/247) (χ2=16.057, P<0.001). The incidence of SIP in infants with BW 400～749 g was 13.0％(14/108), which is higher than infants with BW 750～999 g (3.4％, 8/236) (χ2=11.343, P=0.001). Multivariate Logistic regression analysis showed that twins (OR=4.153, 95％CI 1.392～12.384, P=0.011), umbilical veins catheterization (OR=15.942, 95％CI 1.026～247.789, P=0.048) and ibuprofen use within 3 days after birth (OR=15.387, 95％CI 1.519～155.883, P=0.021) were independent risk factors of SIP. Conclusion The smaller the GA and BW, the higher the incidence of SIP. Twins,umbilical veins catheterization and ibuprofen use early after birth may be independent risk factors of SIP.

Objective:To examine the effect of surgical treatment in children with pulmonary artery sling and the surgical treatment strategy.Methods:Relevant data of 110 children with pulmonary artery sling admitted to the Department of Cardiac Surgery, Children′s Hospital Affiliated to Shandong University from February 2017 to July 2022 were retrospectively analyzed. There were 55 males and 55 females, aging ( M(IQR)) 9.0 (10.6) months (range: 1 to 96 months). The weight was 7.8 (3.5) kg (range: 2.5 to 25.0 kg). Of the 110 patients, 108 had different degrees of tracheal stenosis and 2 had normal trachea. Left pulmonary artery transplantation and tracheoplasty were performed in 78 patients. Left pulmonary artery transplantation was performed in 30 patients (11 in our hospital and 19 in other hospitals) due to the lack of an early tracheoplasty technique, in which 24 patients needed stage Ⅱ tracheoplasty due to obvious respiratory symptoms and limited activity endurance, and 6 cases did not intervene. Two children with normal trachea only underwent left pulmonary artery transplantation. Results:Among the 78 children who underwent surgery in the same period, 70 cases recovered smoothly after surgery, of whom respiratory symptoms were significantly reduced or disappeared during the 1 to 65 months follow-up, with similar activity endurance to normal children of the same age. Eight cases died, including 4 cases of postoperative multi-drug resistant bacteria infection, died from tracheal anastomotic opening or septic shock, 1 cases with severe congenital heart disease died from postoperative low cardiac output syndrome difficult to correct, 1 case died from blood pressure could not be maintained due to the compressed left pulmonary artery after transplantation, 2 cases of postoperative digestive system diseases (adhesive intestinal obstruction, gastrointestinal bleeding, etc.). The 24 patients in the staging group were followed for 1 to 84 months. All patients needed stage Ⅱ tracheoplasty due to respiratory symptoms and decreased endurance to activity. Eight cases of the non-intervention tracheal group were successfully separated from the ventilator, cured and discharged in a short period of time.Conclusions:Most children with pulmonary artery sling have tracheal stenosis. Children with low degree of tracheal stenosis and inconspicuous respiratory symptoms can only undergo left pulmonary artery transplantation by lateral thoracotomy. For patients combined with severe tracheal stenosis or obvious respiratory symptoms, a simultaneous left pulmonary artery transplantat and tracheoplasty is recommended.

Objective:To examine the treatment strategy of congenital tracheal stenosis associated with non-vascular ring cardiac malformations.Methods:This is a retrospective case series study. Clinic data from 24 children with tracheal stenosis who underwent surgical treatment in the Department of Cardiac Surgery, Children′s Hospital Affiliated to Shandong University from February 2017 to March 2023 were retrospectively collected. There were 16 males and 8 females, aged ( M(IQR)) 6.5 (19.6) months (range: 2.2 to 66.3 months) and weighted 5.95 (4.76) kg (range: 3.2 to 20.0 kg). All patients had obvious respiratory symptoms. Eighteen patients underwent cardiac malformation correction and tracheoplasty at the same time (simultaneous group). Six patients in the staged operation group were treated with cardiac malformation correction in the first stage operation and tracheoplasty in the second stage operation due to missed diagnosis or delayed diagnosis of tracheal stenosis or no condition for tracheoplasty. Slide tracheoplasty was used to correct tracheal stenosis in both groups. The recovery of the children was followed. Wilcoxon sign rank test was used for comparison between the two groups. Results:There was no death during the perioperative period and hospitalization. In the simultaneous group, 1 case with delayed chest closure underwent bedside chest closure after 52 hours, 2 cases were intubated again after operation, and 1 case was implanted with an endotracheal stent. The duration of mechanical ventilation was 40.5 (39.6) hours (range: 19.0 to 438.8 hours). In the staged group, there was 1 case of re-intubation after operation, combined with left vocal cord paralysis and respiratory multidrug-resistant bacterial infection ( Acinetobacter baumanii). One patient underwent 3 times of bronchoscopic balloon dilatation of the right middle bronchus, and heart rate returned to normal range. The duration of mechanical ventilation was 19.0 (21.4) hours (range: 17.1 to 96.7 hours). During follow-up, a patient in the simultaneous group was prone to respiratory infection and had good exercise tolerance, 1 case in the staged group still had sputum stridor in the throat 3 months after the operation, and symptoms improved significantly 6 months after the operation. The other children didn′t have obvious respiratory symptoms. Conclusions:The diagnosis of tracheal stenosis may be delayed or missed when tracheal stenosis is complicated by non-vascular ring cardiac malformations. One-stage correction of tracheal stenosis and cardiac malformation can achieve a good outcome.