Objective To investigate the characteristics of epitope distribution on HLA class Ⅰ antigen in the regular platelet donors from Nanjing area and establish the HLA epitope database for regular platelet donors.Methods High-resolution HLA typing was per-formed using Sanger method for the blood samples from 649 regular platelet donors in Nanjing area.The polymorphism of HLA antigen epitopes corresponding to the high-resolution HLA typing results was analyzed using the HLA Eplet Registry website.The frequencies of allele frequencies,HLA haplotype,and HLA antigen epitope were calculated by using the direct counting method.Results Among the 649 regular platelet donors,38 HLA-A alleles were detected,corresponding to 36 HLA-A epitopes,and the higher frequencies were 79GT,144K and 138MI.Seventy-three HLA-B alleles were detected corresponding to 35 HLA-B epitopes,and the higher frequencies were 131S,69TNT,and 80N.Sixty-four HLA class Ⅰ antigen epitopes were detected,in which the higher frequencies were 79GT,131S,and 144K.Conclusions The distribution of HLA antigen epitopes in the regular platelet donors of Nanjing area exhibited u-nique polymorphic characteristics.The epitope matching strategies should be established based on the distribution characteristics of HLA antigen epitopes,which may expand the range of available donors and reduce the incidence of platelet transfusion refractoriness.

Objective To investigate the influence of limonin on the malignant biological behavior of non-small cell lung cancer (NSCLC) cells by regulating the protein tyrosine kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway. Methods CCK-8 method was applied to detect the survival rate of A549 cells treated with different concentrations of limonin (0, 5, 10, 25, 50, 75, 100 μmol/L). A549 cells were separated into normal culture (NC) group, low-dose limonin group (treatment with 10 μmol/L limonin for 24 h), medium-dose limonin group (treatment with 25 μmol/L limonin for 24 h), high-dose limonin group (treatment with 50 μmol/L limonin for 24 h), coumermycin A1 group (treatment with 10 μmol/L JAK2 activator coumermycin A1+50 μmol/L limonin for 24 h), and AG490 group (treatment with 10 μmol/L JAK2 inhibitor AG490+50 μmol/L limonin for 24 h). Clone formation assay was applied to detect the clones of each group of cells. Transwell assay was applied to detect cell migration and invasion, and flow cytometry was applied to detect apoptosis. Western blot analysis was applied to detect the protein expression levels of JAK2, p-JAK2, STAT3, p-STAT3, E-cadherin, N-cadherin, and vimentin in each group. Results The viability of A549 cells decreased significantly in a limonin concentration-dependent manner (P < 0.05), with IC₅₀ of 45.16±1.66 μmol/L. Concentrations of 10, 25, and 50 μmol/L were selected for subsequent experiments. The numbers of clones, migration, and invasion of A549 cells and the protein expression levels of IL-6, p-JAK2, p-STAT3, N-cadherin, and vimentin in the low-, medium-, and high-dose limonin groups significantly decreased, compared with those in the NC group, and the apoptosis rate and E-cadherin protein expression significantly increased (P < 0.05). The JAK2 activator coumermycin A1 attenuated the ability of limonin to inhibit the proliferation, migration, invasion, and other malignant biological behavior of A549 cells and attenuated the apoptosis ability. The JAK2 inhibitor AG490 enhanced the ability of limonin to inhibit the proliferation, migration, invasion, and other malignant biological behavior of A549 cells and enhanced the apoptosis ability. Conclusion Limonin can inhibit the malignant biological behavior of NSCLC cells, such as proliferation, migration, and invasion, by inhibiting the JAK2/STAT3 pathway.

Objective: To explore the growth characteristics of rat calvaria by detecting the calvaria of SD rats in different periods. Methods: The calvaria of SD rats at 1 , 4 , 7 , 10 , and 12 weeks from the same littermate were selected (3 rats per week) . Real⁃time PCR and Western blot techniques were used to detect the expression of focal adhesion kinase ( FAK) Ⅳ phosphatidylinositol 3 kinase/protein kinase B ( PI3K/AKT ) signal pathway in the calvaria , and the role of FAK⁃PI3K/AKT in the growth and development of the calvaria was analyzed by correlation. Results: The increase of brain volume and the thickness of calvaria increased synchronously , the expression of FAK was positively correlated with the changes of meridians , and the expression of FAK was positively correlated with the expression of PI3K/AKT. Conclusion The expression of FAK is related to the growth and development of rat skull. FAK plays a role in calvaria by activating PI3K/AKT signal pathway. FAK may be used as a marker of rapid skull growth and development , which provides a basic theoretical basis for the timing of clinical skull defect repair and treatment.

Objective:To investigate the role and mechanisms of fibulin-1 in senescence-related calcification of rat vascular smooth muscle cells induced by high-concentrationphosphate treatment.Methods:From September 2020 to September 2021, rat primary vascular smooth muscle cells were extracted from the thoracic aorta and abdominal aorta of 10 male SD rats aged 6 to 8 weeks.Phosphate(2.5 mmol/L Pi)was used to stimulate the calcification of vascular smooth muscle cells(VSMCs)in a model of stress-induced senescence-related calcification.Cellular senescence was assessed by SA-β-gal staining.Cellular calcification was determined by alizarin red staining and quantification of calcium deposition.Phenotypic transformation indexes and the expression of fibulin-1 during the process of calcification were detected by Western blot.The expression of fibulin-1 in primary rat vascular smooth muscle cells was knocked down by siRNA, the expression of pSmad3 was detected by immunofluorescence, and the effects of fibulin-1 on phenotypic transformation indexes of smooth muscle cells were detected by Western blot.The cells were cultured with recombinant fibulin-1 while transforming growth factor beta(TGF-β)inhibitor A83-01 and pSmad3 inhibitor SIS3 were also added.The senescence and calcification indexes of smooth muscle cells were detected by Western blot.Results:In the stress-induced aging model with phosphate stimulation of calcification in rat VSMCs, the expression of fibulin-1 was up-regulated( t=11.20, P<0.01), the expressions of MHC and SM22α was down-regulated( t=7.97, P<0.01; t=10.27, P<0.01), and the expression of osteoblastic phenotype markers OPN and Bmp2 and senescence marker P53 was up-regulated( t=4.79, P<0.01; t=9.56, P<0.01; t=14.07, P<0.01). Knockdown of fibulin-1 attenuated the degree of senescence and calcium deposition in VSMCs( t=12.90, P<0.05)and decreased the expression of OPN, Bmp2 and P53( t=5.92, P<0.05; t=10.15, P<0.01; t=8.28, P<0.01), at the same time, and TGF-β and pSmad3 expression was inhibited( t=12.90, P<0.01; t=7.46, P<0.01). After the addition of TGF-β/ smad3 pathway inhibitors, the stimulatory effect of recombinant fibulin-1 on phenotypic transformation and senescence protein expression inVSMCs was significantly reduced( t=4.52, P<0.01; t=9.82, P<0.01; t=3.85, P<0.05). Conclusions:Fibulin-1 can promote aging-related calcification of vascular smooth muscle cells through the TGF-β/smad3 signaling pathway.

Esophagogastric variceal bleeding (EVB) is a common fatal complication secondary to cirrhotic portal hypertension. Terlipressin can shrink visceral blood vessels, reduce portal pressure, and increase renal perfusion and is mainly used in the treatment of EVB. This article summarizes the mechanism of action and clinical effect of terlipressin in the treatment of EVB in liver cirrhosis and points out that if conditions permit, terlipressin combined with endoscopic variceal ligation should be the preferred treatment method for EVB.

Objective To evaluate the effect of an emollient containing Prinsepia utilis Royle oil extracts and other extracts on clinical symptoms and disease recurrence in children aged 2-12 years with atopic dermatitis (AD) in the remission period.Methods A multicenter,randomized,parallel-group,controlled clinical trial was conducted from December 2017 to September 2018.A total of 297 children aged 2-12 years with moderate AD were enrolled from 5 hospitals in China,and randomly divided into the test group (148 cases) and control group (149 cases).In the acute stage,the two groups were both topically treated with mometasone furoate cream once a day on the skin lesions,and with an emollient containing Prinsepia utilis Royle oil extracts and other extracts twice a day throughout the whole body for 2-4 weeks.The children would be enrolled into the remission stage if their Investigator's Global Assessment (IGA) score was ≤ 1 at following visits.In the remission stage,the test group was only topically treated with the emollient twice a day throughout the whole body,while mometasone furoate cream and the emollient were both withdrawn in the control group.At weeks 4,8 and 12 in the remission stage,the recurrence of AD,eczema area and severity index (EASI),children's dermatology life quality index (CDQOL) and adverse events were evaluated.Statistical analysis was carried out with SAS 9.4 software by using t test for comparison of normally distributed continuous data between two groups,chi-square test for comparison of unordered categorical data,Kaplan-Meier method for analysis of survival rates,Cox regression analysis for evaluating the effect of different therapies on AD recurrence in children in the remission stage,and Logistic regression analysis for analysis of odds ratio (OR) of EASI or CDQOL at week 4 in the remission stage between the test group and control group.Results Of the 297 children with AD,31 breached the clinical trial protocol,and 266 were included in the per protocol set (PPS),including 132 in the test group and 134 in the control group.In the PPS,114 and 106 patients completed the follow-up in the test group and control group respectively,and the recurrence rate was significantly lower in the test group (47,41.23％) than in the control group (84,79.25％;x2 =32.96,P ＜ 0.001).The time to recurrence was significantly longer in the test group(61.99 d ± 2.80 d)than in the control group(39.17 d ± 2.54 d,t =6.03,P ＜ 0.001),and the recurrence risk was significantly lower in the test group than in the control group (Log rank test,x2 =32.02,P ＜ 0.001).After adjustment for age and gender,Cox regression analysis showed that the recurrence risk in the test group was 0.35 times that in the control group (HR =0.35,95％ CI:0.24-0.51,P ＜ 0.01).At week 4 in the remission stage,the EASI score at P50-P75 and P75-P100 in the test group were 0.42,0.25 times that in the control group respectively (95％ CI:0.20-0.86,0.12-0.54 respectively;P =0.02,＜ 0.01respectively).Moreover,the CDQOL score at P75-P100 in the test group was 0.33 times that in the control group (95％ CI:0.17-0.65,P ＜ 0.01).No significant difference in the incidence of adverse events was observed between the two groups (P ＞ 0.05).Conclusion Maintenance treatment with the emollient containing Prinsepia utilis Royle oil extracts and other extracts can markedly reduce the recurrence risk in AD children,improve clinical symptoms,and enhance the quality of life.

Midbrain periaqueductal gray ( PAG) is the midline structure of the brain and plays a crucial role in the regulation of a series of physiological functions, emotions and motivated behaviors. In recent years, with the developments of functional tracer technology, genetic technology and neuroimaging technology, the understandings of physiological and psychological functions of PAG have been expanded. The recent research progresses of PAG in the neural circuits in regulation of behaviors, emotions and the related mechanisms are reviewed.

Objective To analyze the CT and MR imaging characteristics of the subcutaneous and organ lesions of uvenile xanthogranuloma (JXG) in children.Methods The CT and MR images of the head,chest,abdomen and subcutaneous soft tissue of 11 patients (6 females and 5 males,aged 2 months-12 years old,median age 8 months old) with JXG from 2006 to 2018 were collected and retrospectively analyzed.Eight patients underwent CT scan and/or contrast-enhanced CT scan,and 7 patients underwent MRI and/or MRI and contrast-enhancement MRI.Results In 11 cases of JXG,5 cases were classified as simple skin type,involving the skin and deep soft tissue in the right posterior neck,the right frontal and parietal parts of the head,the nose root,and the fat layer and the deep muscle in the extremities.The lesions were the subcutaneous soft tissue nodules or mass showing isodensity on CT or high signal on T2WI.And the other 6 cases were classified as systemic type whose lesions involved one or more than one part except the skin.In the 6 cases,3 cases involved the central nervous system,including the pituitary gland,cerebral hemisphere,cerebellum,brain stem,basal ganglia,thalamus,optic chiasma and spinal cord.The lesions of the pituitary gland were manifested as pituitary enlargement,thickening of the pituitary stalk,and disappearance of high signal on T1WI in the neurohypophysis.The brain parenchyma,the spinal cord and the optic chiasma showed multiple flake or nodular masses,which were iso/low signal on T1WI,low / high signal on T2WI,diffusion restricted,and enhanced markedly.Two cases involved lung tissue,and their lesions showed diffuse or scattered multiple cloudy shadow,reticular opacity,strip and small nodule shadow on chest CT.There was pleural effusion in one case.Two cases involved the livers and 2 cases involved the kidneys,showing multiple nodules with low signal on T2,WI and slight enhancement.One case involving the pancreas showed a low density in the head of the pancreas and slight enhancement on CT.One case involving the greater omentum,chest wall and peritoneum showed thickening of the corresponding tissues on CT.Conclusions Subcutaneous and systemic lesions of JXG show some characteristics on CT and MRI images.The main manifestations are single or multiple nodulars,which are diffusely distributed and vary in size in subcutaneous soft tissue and various organs.The combination of imaging appearances,clinical manifestations and biopsy has important clinical significance for the correct diagnosis and evaluation of JXG.

Objective To investigate the in-stent restenosis after vertebral artery ostium stenting (VAOS),and to determine the risk factors for in-stent restenosis. Methods Respective analysis of clinical data of 775 cases received VAOS in Xuan Wu Hospital of Capital Medical University from Jan. 2006 to Dec. 2012. Severe stenosis of vertebral artery ostium were diagnosed by DSA,and followed-up by ultrasound. The risk factors were assessed by COX analysis for in-stent restenosis ≥50%. Results This study included 775 patients. Surgical success rate was 99. 87%(n=774),technique success rate was 99. 48%(n=771 ). Two patients had cerebral hemorrhage after operation,one of them was dead. Four patients had cerebral infarction. The mean follow-up period was 12 months. The restenosis rate was 35. 89%(234/652 ). 79. 91% of restenosis occurred within 12 months after operation. COX analysis showed the vessels diameter after stenting was the independent predictors of in-stent restenosis (P<0. 01). The in-stent restenosis rate of drug-eluting stents was lower than metal-bare stents (HR 0. 532,95%CI 0. 397-0. 713,P<0. 01). Conclusion The in-stent restenosis was peculiarly prone to the smaller vessels diameter after VAOS. Drug-eluting stents were superior to metal-bare stents in preventing in-stent restenosis.

Objective To analyze the chemical constituents in fruits of Capparis spinosa by GC-MS analysis .Methods The compounds were extracted from Capparis spinosa by 70% ethanol after smash .After extraction by petroleum ether (boil-ing range was from 60 to 90 ℃) ,the compounds were analyzed by gas chromatography with a VF-5ms capillary column .The column was heated up from 80 ℃ to 300 ℃ ,15 ℃/min ,then maintained for 10 minutes ,vaporization temperature was 250 ℃ , carrier gas was Helium and the flow rate was 1 ml/min .The detection was carried out by mass spectrometry using electron im-pact ion source with 70 eV ionization voltages .The ion source temperature was 200 ℃ and split ratio was 30∶1 .The injection volume was 1 .0 μl .The content of the compounds was determined with peak area normalization method .Results 53 chroma-tographically peaks were separated and 48 chemical constituents were identified including hexadecanoic acid (21 .82% ) ,octade-canoic acid (7 .49% ) ,oleinic acid (42 .93% ) and monoolein (2 .39% ) .These chemical constituents were mainly saturated fatty acid ester ,unsaturated fatty acid ester and alkanes .Conclusion The method is rapid ,accurate and sensitive with the usage of GC-MS ,which could be used to identify main chemical constituents of Capparis spinosa in fruit .