Objective:To observe the expression of BTLA on T cells during activation and further analyze its inhibitory effects on T cell activation in different phases.Methods: T cells from PBMC were enriched by negative selection using magnetic beads.Expression of BTLA,CTLA-4 and PD-1 on freshly isolated human T cells and kinetics expression of BTLA,CILA-4 and PD-1 on CD3 mAb stimulated T cells were examined by flow cytometry.T cells were stimulated by anti-CD3 mAb combined with anti-CD28 mAb in the presence of anti-BTLA mAb 8H9,then T cell proliferation was tested by MTT assay in the different culture time.Immature DCs were generated from monocytes cultured in the medium containing GM-CSF and IL-4, and further driven to maturation by anti-CD40 mAb.Expression of HVEM on DCs was measured by flow cytometry.T cells were co-cultured with DCs in the presence of soluble 8H9 or anti-HVEM antibody to block HVEM-BTLA interaction,T cell proliferation was measured by MTT assay.Results:Freshly isolated T cells exhibited high levels of BTLA expression, but not CTLA-4 and PD-1.After T cell activation, BTLA expression decreased on first 2 days, with rapidly increasing to high levels.Unlike BTLA, expression of CTLA-4 and PD-1 was gradually increased during T cell activation.8H9 significantly inhibited the proliferation of T cell stimulated by CD3 mAb and CD28 mAb.8H9 could still exhibit inhibitory effect on T cell proliferation after 24 h or 48 h of preactivation by CD3 mAb plus CD28 mAb stimulation.HVEM was highly expressed on immature DCs, and down-regulated on mature DCs.Blockade of BTLA by soluble 8H9 or anti-HVEM antibody enhanced DC-mediated T cell proliferation within 48 h.Conclusion: BTLA signal enhances the threshold of T cell activation and plays importantly negative regulatory role in the initiation and early phase of T cell activation.

Objective: To develop a molecular screening test for genetic defects on hearing loss related genes has significant impacts on early identification of hereditary hearing loss and genetic susceptibility to aminoglycoside ototoxicity. Early identification of pre-lingual hearing loss is very important for patient's language development, academic achievement, and social skill. Two common mutations, the 235delC in GJB2 gene and the mutation A1555G in mitochondrial DNA, are included in the newly developed screening panel for Chinese population. Methods: A molecular genetic assay, based on fluorescent labeled multiplex PCR and automatic DNA fragment analyzing techniques, was developed to detect both mutations simultaneously. Results: This assay was able to detect both mutations from patient's samples, and pooled DNA tests, as well as suitable to detect mutation from the DNA extracted from dried blood spot and buccal swab. Conclusion: This assay could be a useful tool for newborn screening and carrier screening for the hereditary hearing loss for the Chinese population.

The stage of impaired glucose regulation is a necessary stage in the development of diabetes.Nearly a quarter of patients without diabetes intervention develop into diabetes within four to seven years without intervention.Therefore,the intervention of pre-diabetes treatment in the prevention and control of diabetes is of great significance.This study was to investigate the effect of Tangnaikang granule at the early stage of type 2 diabetes mellitus.The age,course of disease,sex,symptom score of TCM were recorded,so were the blood lipid,blood glucose,liver and kidney function,electrocardiogram were recorded.Under the condition of low sugar and low salt diet and healthy exercise,the patients were divided into the control group (n--31) and the treatment group (n=43).The patients in the double-blind control group were treated with Kangnaikang granule and Tangnaikang granule to treat 74 patients with type 2 diabetes mellitus three months.Three months after the detection of fasting and postprandial blood glucose,glycosylated hemoglobin,glucose tolerance,insulin tolerance and other indicators to examine the efficacy of both.Tangnaikang granules were superior to placebo in terms of effective rate of treatment,TCM symptom score and blood sugar recovery rate,and taking Tangnaikang granule without liver and kidney function and other injuries.Tangnaikang is safe and effective for pre-diabetes patients.

Specimens of skin, skeletal muscle, adipose tissue, brain, peripheral nerve and catilage were taken from 6 cadavers (divided into 2 groups) perfused and embalmed with formalin. 90 pieces of the above-mentioned tissues were taken in different times and the paraffin sections were stained by routine HE. Electron microscopic observation was made from some skin and skeletal muscle specimens, and histochemical methods were used to demonstrate DNA, RNA, keratin(-SH, -SS), fatty acids and calcium in some skin materials. Loyez stain method for myelin sheath was applied to some brain and nerve sections.1. Skin tissue: In both group A and B, 83.3% and 78.9% of skin materials were appraised as grade I and II (excellent and good) respectively. The morphological changes appeared mainly as karyopyknosis of the epidermal cells and shrinkage of the secretory part of the sweat glands. Most of the skin tissues of the corpses perfused 3～4 hrs after death were well preserved, only 8.3% being in grade Ⅲ (moderate). On the other hand, 43% of skin materials taken from those perfused 30 hrs after death were in grade Ⅲ. Ultrastructurally, the shape of the epidermal cells and their adhensions still remained intact, but karyopyknosis or chromolysis and autolysis of most of the cytoplasmic membraneous structures were found. In those cadavers preserved for less than one year, there was no evidence of progressive changes in microscopic and submicroscopic structures of the skin. The tonofillaments and desmosomes were still kept well in those specimens preserved for many years, and their presence may be the cause of maintaining cell integrity. Histochemically, mercapto-reaction of keratin was kept stable and found to be well shown even in old samples of skin, but the disulfide reaction usually showed marked fading with time, probably due to exofolition of the stratum corneum. However, this reaction was well shown in the hairs and not weakened for many years.2. Fat tissue: Hydrolysis of lipid substances of subcutancous fat and sebaceous gland, as shown by the presence of fatty acid, was found in the corpses preserved even as short as half a year.3. Skeletal muscle: The preservation state of the skeletal muscle was also dependent on the complete perfusion in time. The structure of the muscle fibers was generally worse preserved than that of the skin as shown in severe myolysis. Since stagnation of the small blood vessels frequently accompanied myolysis, obstruction of microcirculation which prevented the permeation of fomalin into the muscle tissues may be the cause of rapid myolysis. The ultrastructural changes of the muscle fibers were the same as those of the epidermal cells. In all specimens even as old as 30 years, two types of myofillaments were well preserved, and bands could be seen distinctly.4. Nervous tissue: The brain tissues were well preserved in those corpses that were perfused shortly after death, but the peripheral nerves usually showed severe autolysis, obviously it is due to the same reasons as those for myolysis.5. Cartilage: The structural changes of cartilage after death were slow and all specimens of cartilage were well preserved.

Objective To investigate the effects and the mechanism of sorafenib on hepatocellular car-cinoma growth and vasculogenic mimicry (VM)in mice.Methods A subcutaneous implantation mouse model of human hepatocellular carcinoma (HCC)HepG2 cells were established.Mice inoculated with HepG2 cells were randomly divided into the treatment group (sorafenib 30 mg·kg -1 ·d -1 )and the control group by using of paired comparison method.Growth of established tumor xenografts was monitored at least twice weekly by vernier caliper measurements.VMwas assessed by immunohistochemical assay and periodic acid schiff reaction (PAS)histochemical double-staining.The expressions of HIF-1 α,VEGFA,VEGFR-1 and MMP-2 in tumor tissues were also assessed by immunohistochemical assay,Western blotting and real-time quantitative PCR. Results The tumor volume in the sorafenib group was obviously decreased compared with the control group (809.69 mm3 ±208.71 mm3 vs 1 678.00 mm3 ±31 3.29 mm3 ),with a statistically significant difference (t =6.1 03,P =0.030).Haematoxylin and eosin (HE)staining showed that tumor tissues treated with sorafenib were characterized by obvious necrosis,but there were not the same cases in the control group.Sorafenib group significantly reduced the number of tumor functional vessel in HepG2 xenografts compared with the control group,as assessed by tumor vasculature uptake of DiOC7 (4.77 ±0.1 5 vs 8.44 ±0.68,t =9.1 92,P =0.01 3).The number of VMwas significantly decreased by sorafenib (1 .04 ±0.46 vs 2.66 ±0.42,t =4.51 0, P =0.041 ).Relative to controls,CD31 -positive vessels decreased after treatments (3.42 ±0.1 0 vs 1 .26 ± 0.1 4),with a statistically significant difference (t =21 .580,P =0.002).Compared with the control group, the protein levels of HIF-1 α(0.65 ±0.03 vs 1 .00 ±0.00),VEGFA (0.51 ±0.02 vs 1 .00 ±0.00), VEGFR-1 (0.45 ±0.04 vs 1 .00 ±0.00)and MMP-2 (0.69 ±0.02 vs 1 .00 ±0.00)were significantly decreased in the sorafenib group (t =1 9.650,P =0.003;t =40.493,P =0.000;t =23.429,P =0.002;t =26.071 ,P =0.002).Compared with the control group,the mRNA levels of HIF-1 α(0.78 ±0.05 vs 1 .00 ±0.00),VEGFA (0.52 ±0.05 vs 1 .00 ±0.00),VEGFR-1 (0.45 ±0.02 vs 1 .00 ±0.00)and MMP-2 (0.71 ±0.02 vs 1 .00 ±0.00)were also significantly decreased in sorafenib group (t =6.840,P =0.021 ;t =27.71 0,P =0.001 ;t =62.740,P =0.000;t =23.850,P =0.002).Conclusion Sorafenib can inhibit the tumor growth and VMchannels formation,which may be related with the HIF-1 αand VEGFA /VEGFR-1 signa-ling pathway.

Objective: To prepare anti-VSIG4 monoclonal antibodies and characterize their biological functions.Methods: BALB/c mice were immunized with transfected cell line (L929/VSIG4L) as immunogen.The spleen B cells of the mice were fused with SP2/0 and hybridoma cells were screened with transfected cell line (L929/VSIG4) by FCM.After acquisition of the hybridomas secreting anti-VSIG4 mAb,their biological activities were investigated by indirect immunofluorescence,Western blot,competitive inhibition test,and MTT assay.Results:Two stable hybridomas,9A7 and 9D5 were obtained,which could continuously secrete specific anti-VSIG4 monoclonal antibodies.The following biological activity studies showed that these monoclonal antibodies could recognize the natural VSIG4 expressed on the macrophages and several cancer cell lines,such as Jurkat,THP-1 and H446.Furthermore,they could block the inhibitory effects of VSIG4 on proliferation of T cells in vitro.Conclusion: Two hybridomas secreting anti-VSIG4 monoclonal antibodies have been established.These monoclonal antibodies provide useful tools for further studying VSIG4's biological function and its unknown receptor.

Objective:To summarize the midterm to long-term outcomes and experiences of endovascular treatment (ET) of spontaneous isolated superior mesenteric artery dissection (SISMAD).Methods:The clinical data of 31 SISMAD patients from Jan 2011 to Dec 2019 treated with ET was retrospectively analyzed.Results:Successful ET was achieved in 29 patients with a technical success rate of 93.5%. A total of 36 self-expandable bare stents were planted in 28 patients and plain old balloon angioplastry (POBA) was performed in 1 patient. Abdominal pain disappeared within 24 hours in 89.3% of the patients after stenting. The rate of perioperative complication was 3.2%. There was no SMA dissection rupture bleeding, nor perioperative death occurred. The mean follow-up time was 53.5 (range, 6 to 110) months. There was no dissecting aneurysm formation, no SMA rupture and bleeding, and no stent rupture during the follow-up. The post ET 1-year, 3-year, and 5-year free from reintervention rate were 100%, 100%, and 91.7%, respectively.Conclusions:ET for SISMAD is safe and effective with satisfactory perioperative and midterm to long-term outcomes.

Objective To study the critical thinking situation among different nationalities undergraduate nursing students and explore its training in basic nursing teaching , so as to provide the evidence for the teaching reform .Methods Routine teaching methods and case teaching methods were respectively used in ‘basic nursing ’ teaching among different nationalities undergraduate nursing students , and the Critical Thinking Disposition Inventory ( CTDI-CV ) was used to investigate their critical thinking abilities . Results Among 69 undergraduate nursing students in the study , gender:55 female students (79.7%),and 14 male students(20.3%), nationality:37 minorities(53.6%) ,and 32 Han nationality(46.4%),and more than 91.3%of students approved the importance of critical thinking training .Before and after the study , critical thinking total scores of different nationalities respectively were (251.21 ±22.94) and (256.65 ±31.46), both less 280 ,and the dimensions the score of which were more than 40 were self-confidence and thirst for knowledge . The critical thinking total scores of male students before and after the study were respectively higher than those of female students [(254.28 ±34.20) vs (250.87 ±21.68), (289.71 ±54.62) vs (252.91 ±25.89)].The dimensions the score of which were more than 40 in male students respectively were analysis , self-confidence , thirst for knowledge , and systemic capacity , and that in female students only was self-confidence dimension before and after the study .Conclusions Undergraduate nursing students have high approval for the training of critical thinking , however their critical thinking level is in medium .Critical thinking total score of male students is higher than that of female students , and male students have positive tendency of critical thinking after the study,which show that reformed teaching methods can significantly improve the male students ’ critical thinking while no significant effectiveness is found in improving the entirety critical thinking of undergraduate nursing students.More explore and lots work need to do for the critical thinking training in the future .

OBJECTIVETo study the effects of polypeptide extract from scorpion venom (PESV) on immune escape of Lewis lung carcinomas (LLC) and its mechanism.

METHODForty C57BL/6J mice were inoculated with LLC cells suspension (1 x 10(7) cells/ mL) in right armpit subcutaneously. The tumor-bearing mice were randomly divided into two groups: the control group and the PESV group. PESV was intragastrically subjected to the mice of the experimental group for 18 days. The tumor volume and tumor inhibitory rate were determined. The expression levels of VEGF,TGF-beta1 and IL-10 in tumor microenvironment were determined by immunohisto-chemistry-staining and ELISA. Surface co-stimulatory molecules CD80 and CD86 of tumor infiltrating dendritic cells (DC) were determined by immunohistochemistry-staining and flow cytometry.

RESULTThe growth inhibitory rate of PESV was 56. 60%. The expression levels of VEGF,TGF-beta1 and IL-10 were decreased in tumor and serum, while the expression of co-stimulatory molecules CD80 and CD86 on DC were increased in tumor. Compared with the control group, the differences were all significant (P < 6.05).

CONCLUSIONPESV was effective in recovering immuno-surveillance and intervening immune escape of lung cancer through multi-pathway. And its effects might be attained by decreasing the level of VEGF, TGF-beta1 and IL-10 in tumor microenvironment and increasing the expression of co-stimulatory molecules CD80 and CD86 on DC.

Animals ; B7-1 Antigen ; immunology ; B7-2 Antigen ; immunology ; Carcinoma, Lewis Lung ; drug therapy ; immunology ; Disease Models, Animal ; Humans ; Interleukin-10 ; immunology ; Lung Neoplasms ; drug therapy ; immunology ; Male ; Mice ; Mice, Inbred C57BL ; Peptides ; administration & dosage ; immunology ; isolation & purification ; Scorpion Venoms ; chemistry ; immunology ; Tumor Escape ; drug effects

Objective Through the community health management of hypertension , to determine the input and output , change the knowledge , attitude, practice ( KAP) of hypertension patients and put forward the reasonable suggestions of antihypertensive treatment in order to reduce hypertensive patients 'medical expenses . Methods By random cluster sampling method , we selected 809 hypertension patients , over 18 years old, from two communities in Urumqi .414 patients were in the management group and 395 patients were in the control group.The health management the cost benefit was analyzed by the change of patient 's KAP, medical expenses and so on.Results For the knowledge and behavior changes , 17.94% patients in the management group and 3.04%patients in the control knew the classification of blood pressure (Z=-5.688,P=0.000).The master of normal range of blood pressure (Z=-11.927,P=0.000), knowledge of obesity (χ2 =15.608,P=0.000) and knowledge of risk factors (χ2 =-17.677,P=0.000) between groups had significant differences (P<0.01).The total drug expenses of the management group and the control group were (1 583.88 ±981.24) and (1 923.90 ±1 523.81) yuan, respectively;the total hospitalization expenses of the management group and the control group were (10 725.77 ±11 688.26) and (15 327.03 ±10 280.42) yuan, respectively, the differences were significant (t=2.14,2.75, respectively;P<0.05).The costs of patients taking the drug and other costs in the management group were lower than the control group , the differences were significant ( P <0.01 ). Conclusions Hypertension health management can obtain a good cost benefit , reduce hypertension sickness cost, and improve quality of life .