Objectiye To investigate the expression of regulated on activation, normal T-cell expressed and secreted (RANTES) and occluding in brain, and investigate the role of expression of RANTES in the change of blood-brain barrier permeability of rats with acute pancreatitis. Methods 49 Sprague-Dawley rats were randomly divided into 7 groups according to random number table, including sham-operated group (SO) , acute edematous pancreatitis (AEP) 3, 6 h group and acute necrotizing pancreatitis (ANP) 3, 6, 12 and 24 h group. AEP and ANP were induced by retrograde injection of 0. 5% and 5% sodium taurocholate, respectively. RT-PCR, Western blot, and immunohistochemistry methods were performed to assess the expression of RANTES and occludin mRNA and protein in brain tissue. Results The RANTES mRNA expressions in SO, AEP 3, 6 h, ANP 3, 6, 12, 24 h group were 0, 0, 0, 0.36±0.05, 0.47±0.04, 0.65±0.05, 0.83±0.07, respectively; The RANTES protein expressions were 0, 0, 0, 0.42±0. 03, 0. 57±0.04,0.78±0.08, 1.05±0.08, respectively; the values in ANP group were significantly lower than those in SO group and AEP group (P<0.01). The occludin mRNA expressions were 1.21±0.07,1.17±0.07, 1.15±0.08,0.84±0.07,0.77±0.05,0.64±0.09,0.56±0.09, respectively, the occludin protein expressions were 1.18 ±0.08, 1. 16 ±0. 10, 1. 11 ±0. 10, 0. 90 ±0. 03, 0. 65 ±0."05, 0.57 ±0.05, 0.48 ±0.05, respectively, the values in ANP group were significantly lower than those in SO group and AEP group (P< 0.01). The expression of RANTES was positively related to the pancreatic pathologic score (r = 0. 936,P< 0. 001) ; the expression of occluding was negatively related to the pancreatic pathologic score (r = - 0. 943, P < 0.001). The expression of RANTES was negatively related to the expression of occluding (r = -0. 943, P <0. 001). Conclusions The expression of RANTES was progressively increased in the brain tissue in rats with ANP, while the expression of occluding was progressively decreased; RANTES may play an important role in the change of blood-brain barrier permeability via down-regulating the expression of occluding.

Objective:To identify the hub differentially expressed genes(DEGs)of glomerular pathological changes and potential pathways in molecular process of type 2 diabetic nephropathy(DN)based on bioinformatics technology.Methods:The differentially expressed genes of Gene Expression Omnibus(GEO)dataset GSE96804 in DN and normal kidney tissues were analyzed by R 3.6.2 software. DEGs were further assessed by Gene Ontology(GO)function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signal pathway analysis. Subsequently, the hub genes and their associated pathways were analyzed using String 11.0 and Cytoscape 3.7.2 software.Results:A total of 168 DEGs were obtained in the dataset. Among them, seven hub genes were identified, including ALB, FN1, EGF, PTGS2, PLG, KDR, and LOX. Three hub genes, ALB, EGF, PLG, exerted a direct action on glomerulus. GO enrichment analysis of DEGs was mainly manifested in extracellular matrix organization, extracellular structure organization, platelet degranulation and other biological processes, extracellular matrix, secretory granule lumen, platelet alpha granule and other cell components, chaperone binding, copper ion binding, antioxidant activity, and other molecular functions. DEGs mainly regulated metabolic process, which was related to fatty acid degradation signal pathway, exogenous substance metabolism related to CYP enzyme and drug metabolism signal pathway.Conclusion:A bioinformatics analysis of DN from the perspective of glomerulopathy is helpful to understand the potential molecular mechanism of DN and provide reference for further validation.

Objective To observe the relationship of viral load,serum cytokines and tissue damage after severe fever with thrombocytopenia syndrome virus (SFTSV)infection,and to explore the impact of SFTSV levels on tissue injury and prognosis.Methods Twenty-four ambulatory and hospitalized patients who were infected with SFTSV were enrolled between May 2011 and July 2012 at Department of Infectious Disease, First Affiliated Hospital with Nanjiang Medical University. According to their prognosis,they were divided into cure and death group,while 32 healthy blood donators were also enrolled from center blood station in Nanjing as control.The serum SFTSV load was detected using fluorescence quantitative polymerase chain reaction (PCR).The serum T helper (Th)1/Th2/Th17 cytokines in patients with severe fever with thrombocytopenia syndrome (SFTS)were determined dynamically and quantitatively by flow cytometry.The relationships between viral load,cytokines and serum enzymes, white blood cell (WBC),platelet (PLT)counts were analyzed.Comparisons among groups were achieved by rank sum test and correlation analysis among serum cytokines,blood cell counts and tissue damage was done by Spearman correlation test.Results All of the 24 patients showed a positive reaction to SFTSV RNA.The SFTSV loads of 21 cured cases,those of 2 were > 7.0 lg copy/mL,and those of 3 death patients were 6.7 lg copy/mL,8.8 lg copy/mL and 9.8 lg copy/mL,respectively.Serum level of interleukin (IL)-6 (21 .76 pg/mL in day 5 and 7.12 pg/mL in day 7)and IL-10 (14.33 pg/mL in day 5 , 14.13 pg/mL in day 7 and 3.01 pg/mL in day 9)of cured patients were significantly higher than those of healthy controls (IL-6:2.82 pg/mL and IL-10:1 .56 pg/mL)(P <0.05 ).At day 7 and day 9,serum levels of IL-6 of death cases were 137.61 pg/mL and 1 450.83 pg/mL,respectively and serum levels of IL-10 were 50.26 pg/mL and 49.43 pg/mL,respectively.Both of the indicators in the death group were significantly higher than those of cure group (P <0.05 ).However,serum levels of IL-2 and IL-4 were significantly lower than those in healthy control group (P <0.05 ).In the cure group,WBC and PLT counts were lowest during the early course of the disease,and serum alamine aminotransferase (ALT), aspartate aminotransferase (AST ), lactic dehydrogenase (LDH ) and creatine kinase (CK ) were significantly higher than their upper limits of normal.The correlation analysis showed that serum IL-6, IL-10 levels were negatively correlated with PLT count (r=-0.390 and -0.608,respectively;both P <0.01),and positively correlated with SFTSV load (r=0.560 and 0.758,respectively),ALT (r=0.412 and 0.390,respectively),AST (r = 0.686 and 0.764,respectively),LDH (r = 0.633 and 0.677, respectively)and CK (r =0.527 and 0.636,respectively)(all P <0.01 ).Conclusions SFTSV load, IL-6,IL-10 and serum enzyme levels are closely related to the severity of the disease.The inflammatory and anti-inflammatory cytokine storm after SFTSV infection may be involved in the immune pathological injury in patients with SFTS.

Abstract: Objective　To analyze the influencing factors of mother-to-child transmission of hepatitis B virus after combined immunological blockade, and to evaluate the effect of mother-to-child blockade, and to provide a basis for health policies and health interventions for preventing mother-to-child blockade of hepatitis B virus. Methods A total of 11 363 pairs of HBsAg positive pregnant women and their infants aged 7-12 months in Hainan Province from 2015 to 2020 were included in the study. The general situation, the situation of health care and delivery in this pregnancy and perinatal period, the detection of hepatitis B markers, the situation of antiviral therapy, the general situation of mother and infant during delivery and the implementation of blockade measures for mother-to-child transmission of hepatitis B were collected and analyzed. Results Among the 11 363 pairs of HBsAg positive pregnant women and their infants delivered in hospitals in Hainan province from 2015 to 2020, the positive rate of HBsAg in children at 7-12 months after birth was 1.47 %, and the difference in HBsAg positive rate of infants born in different years was not statistically significant (P>0.05). There were no significant differences in the positive rate of HBsAg among children born to pregnant women with different nationalities, educational levels, occupations, delivery modes, delivery places, obstetric operations and perineal laceration, abnormal perinatal period, children with different genders and premature delivery and perinatal (all P<0.05). There was significant difference in HBsAg positive rate among infants born to pregnant women of different ages, the positive rate of HBsAg of infants born to young pregnant women was higher than that of older pregnant women (P<0.05). The rate of antiviral therapy was low in HBeAg positive pregnant women, and the positive rate of HBsAg in their infants was 2.54%, which was higher than 0.83% in HBeAg negative pregnant women (P<0.05). Conclusions Combined immunological blockade with hepatitis B vaccine and hepatitis B immunoglobulin can effectively prevent the mother-to-child transmission of HBV. HBsAg-positive women can give birth at the right age, and HBeAg-positive pregnant women can be treated with antiviral therapy to block mother-to-child transmission, providing the important basis for the formulation of hepatitis B prevention and control strategies and measures.

Objective:To evaluate the role of tumour necrosis factor-α-induced protein 8-like 2 (TIPE2) in the acute lung injury (ALI) induced by endotoxin in mice.Methods:Forty SPF healthy adult male BALB/c mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=10 each) using a random number table method: vehicle plasmid group (VP group), vehicle plasmid plus ALI group (VP+ ALI group), TIPE2 adeno-associated virus overexpression group (T group) and TIPE2 adeno-associated virus overexpression plus ALI group (T+ ALI group). The mice in VP and VP+ ALI groups were injected with empty adeno-associated virus, while the mice in T and T+ ALI groups were intratracheally given adeno-associated virus carrying TIPE interference sequence.Three weeks later, the model of endotoxin-induced ALI was established.Lipopolysaccharide (LPS) 5 mg/kg was intratracheally given in VP+ ALI and T+ ALI groups, and the equal volume of phosphate buffered saline (PBS) was given in VP and T groups.Blood samples were obtained from the abdominal aorta at 24 h after injection of LPS for blood gas analysis, oxygenation index (OI) was calculated, and tumor necrosis factor-alpha (TNF-α) in serum were detected by enzyme-linked immunosorbent assay.The animals were then sacrificed, and lung tissues were removed for examination of pathological changes which were scored after haematoxylin and eosin staining, for calculation of the wet/dry weight ratio (W/D ratio) and for determination of myeloperoxidase (MPO) activity and the expression of TIPE2, phosphorylated c-Jun N-terminal kinase (p-JNK) and nuclear factor kappa B(NF-κB) (by Western blot). Results:Compared with VP group, the lung injury score, W/D ratio, MPO activity and concentration of serum TNF-α were significantly increased, PaO 2 and OI were decreased, expression of TIPE2 was down-regulated and expression of p-JNK and NF-κB was up-regulated in VP+ ALI group ( P<0.05). Compared with VP+ ALI group, the lung injury score, W/D ratio, MPO activity and concentration of serum TNF-α were significantly decreased, PaO 2 and OI were increased, expression of TIPE2 was up-regulated and expression of p-JNK and NF-κB was down-regulated in T+ ALI group ( P<0.05). Conclusion:The down-regulation of TIPE2 expression is involved in the process of ALI induced by endotoxin in mice.

Objective To study the diagnostic values of X-ray,CT and MRI for the hip joint lesion of the patient with ankylosing spondylitis (AS).Methods Totally 180 AS patients underwent the examinations of X-ray,CT and MRI from January 2014 to December 2015,and the hip joint lesion,clinical features and the efficacies of the three imaging techniques were observed.Results The rates for positive findings were 69.44％,71.11％ and 72.22％ respectively by X-ray,CT and MRI,and more than 90％ patients had the lesions occurred at the bilateral hip joints simultaneously.Hip joint lesion of AS had the imaging features of bone marrow and joint space changes,which related to the disease progress closely.Early positive signs consisted of medial joint space narrowing by X-ray,rear joint space narrowing by CT as well as bone marrow fat deposition (BMFD) and bone marrow edema (BME) by MRI.Conclusion X ray,CT and MRI all gain their advantages when used to diagnose the hip joint lesion of AS.X-ray behaves well in early diagnosis of medial joint space narrowing,while have disadvantages in displaying soft tissues and minute structures when compared with CT and MRI.CT displays the minute structure of hip joint clearly,and is not so good at soft tissue resolution as MRI.MRI gains the highest resolution for displaying soft tissues in the three techniques,and can show BMFD,BME and etc which can not be revealed by X-ray and CT.

Objective To evaluate the role of tumor necrosis factor α-induced protein 8-like-2 ( TIPE2) in pyroptosis in macrophage of mice using small interfering RNA ( siRNA) technique. Methods J774A. 1 macrophages of mice were divided into 4 groups ( n=6 each) using a random number table method: non-specific siRNA (Scr-siRNA) group (S group), Scr-siRNA plus LPS∕ATP group (S+LPS∕ATP group ) , TIPE2-siRNA group ( T group ) and TIPE2-siRNA plus LPS∕ATP group ( T+LPS∕ATP group) . The corresponding siRNA and Lipofectamine2000 transfection reagents were added to each group, and transfection was performed for 24-48 h, and in addition LPS 1. 0 μg∕ml was then added, cells were incubated for 5 h, then ATP 5. 0 mmol∕L was added, and cells were incubated for 1 h in S+LPS∕ATP and T+LPS∕ATP groups. Cells were collected to detect the expression of TIPE2, NOD-like receptor familypyrin domain containing 3 (NLRP3), caspase-1, apoptosis-associated speck-like protein containing a caspase activation and recruitment domain ( ASC) and Gasdermin D ( GSDMD) ( by Western blot) . The superna-tant was collected for determination of lactic dehydrogenase ( LDH) activity and concentrations of interleu-kin-1beta (IL-1β) and IL-18 (by enzyme-linked immunosorbent assay). Results Compared with group S, the expression of TIPE2 was significantly down-regulated, the expression of NLRP3, caspase-1, ASC and GSDMD was up-regulated, and the LDH activity and concentrations of IL-1βand IL-18 in supernatant were increased in group S+LPS∕ATP ( P<0. 05) . Compared with group T, the expression of TIPE2 was sig-nificantly down-regulated, the expression of NLRP3, caspase-1, ASC and GSDMD was up-regulated, and the LDH activity and concentrations of IL-1βand IL-18 in supernatant were increased in group T+LPS∕ATP (P<0. 05). Compared with group S+LPS∕ATP, the expression of TIPE2 was significantly down-regulated, the expression of NLRP3, caspase-1, ASC and GSDMD was up-regulated, and the LDH activity and con-centrations of IL-1βand IL-18 in supernatant were increased in group T+LPS∕ATP ( P<0. 05) . Conclusion Application of siRNA technique once again confirms that TIPE2 can inhibit pyroptosis in macrophages of mice.

Objective To evaluate the effects of dexmedetomidine on NLRP3 inflammasome during acute lung injury ( ALI ) induced by blunt chest trauma and hemorrhagic shock-resuscitation in rats. Methods Forty-five SPF healthy male Sprague-Dawley rats, weighing 200-220 g, were divided into 3 groups (n=15 each) using a random number table method: sham operation group (Sham group), ALI group and dexmedetomidine group ( Dex group) . A rat model of ALI was established by blunt chest trauma and hemorrhagic shock-resuscitation. Dexmedetomidine 5 μg·kg-1 ·h-1 was infused into the femoral vein after blunt chest trauma in Dex group. Blood samples were collected from the femoral artery at 6 h after blunt chest trauma for measurement of arterial oxygen partial pressure and concentrations of pro-inflammatory cytokines interleukin-1beta ( IL-1β) and IL-18 in serum ( by enzyme-linked immunosorbent assay ) . The oxygenation index was calculated. The rats were sacrificed and lungs were removed for examination of the pathological changes which were scored and for determination of the expression of NLRP3, caspase-1 and apoptosis-associated speck-like protein containing CARD ( ASC) in lung tissues ( by Western blot) and ac-tivities of lactic dehydrogenase ( LDH) and myeloperoxidase ( MPO) in lung tissues ( by colorimetry) . Re-sults Compared with Sham group, PaO2 and oxygenation index were significantly decreased, the expres-sion of NLRP3, caspase-1 and ASC was up-regulated, the activities of LDH and MPO were increased, and the concentrations of IL-1β and IL-18 in serum were increased in ALI group ( P＜0. 05 ) . Compared with ALI group, PaO2 and oxygenation index were significantly decreased, the expression of NLRP3, caspase-1 and ASC was down-regulated, the activities of LDH and MPO were decreased, and the concentrations of IL-1β and IL-18 in serum were decreased in ALI group ( P＜0. 05) . Conclusion Dexmedetomidine can al-leviate blunt chest trauma and hemorrhagic shock-resuscitation induced ALI in rats, and the mechanism is related to inhibition of NLRP3 inflammasome activation and reduction of inflammatory response.

Objective To evaluate the role of tumor necrosis factor alpha-inducible protein 8 like-2 ( TIPE2) in macrophage pyroptosis in mice. Methods Mouse macrophages J774A. 1 were seeded in 6-cm culture dishes (5 ml∕dish) at the density of 2×105 cells∕ml and divided into 4 groups (n=18 each) using a random number table method: blank vector control group ( C group) , blank vector plus lipopolysaccharide ( LPS)∕ATP group ( C+LPS∕ATP group) , TIPE2 overexpression group ( T group) and TIPE2 overexpres-sion plus LPS∕ATP group ( T+LPS∕ATP group) . Cells were infected with lentivirus without TIPE2 in C and C+LPS∕ATP groups. TIPE2 overexpression stable cell line was constructed in T group and T+LPS∕ATP group. LPS 1. 0 μg∕ml was added and cells were incubated for 5 h, and then ATP 5. 0 mmol∕L was added and cells were incubated for 1 h in C+LPS∕ATP group and T+LPS∕ATP group. Cells were collected for de-tection of the expression of TIPE2, NOD-like receptor protein 3 ( NLRP3) , interleukin-1beta ( IL-1β) and interleukin-8 ( IL-18) by Western blot. Flow cytometry was used to detect the pyroptotic cells, and the per-centage of pyroptotic cells was calculated. The concentrations of tumor necrosis factor-alpha ( TNF-α) , IL-6, IL-1β and IL-18 in cell culture media were determined by enzyme-linked immunosorbent assay. Results Compared with group C, the expression of TIPE2 was significantly down-regulated, the expression of NL-RP3, IL-1β and IL-18 was up-regulated, and the percentage of pyroptotic cells and concentrations of TNF-α, IL-6, IL-1β and IL-18 in cell culture media were increased in group C+LPS∕ATP (P<0. 05). Com-pared with group T, the expression of TIPE2 was significantly down-regulated, the expression of NLRP3, IL-1βand IL-18 was up-regulated, and the percentage of pyroptotic cells and concentrations of TNF-α, IL-6, IL-1β and IL-18 in cell culture media were increased in group T+LPS∕ATP ( P<0. 05) . Compared with group C+LPS∕ATP, the expression of TIPE2 was significantly up-regulated, the expression of NLRP3, IL-1β and IL-18 was down-regulated, and the percentage of pyroptotic cells and concentrations of TNF-α, IL-6, IL-1β and IL-18 in cell culture media were decreased in group T+LPS∕ATP ( P<0. 05) . Conclusion TIPE2 can inhibit macrophage pyroptosis, and the mechanism may be related to inhibiting activation of NL-RP3 inflammasome in mice.

Objective: To evaluate the effect of penehyelidine hydrochloride (PHCD) on tumor necrosis factor α-induced protein 8-like-2 (TIPE2)-Toll-like receptor 4 (TLR4)-myeloid differentiation factor 88 (MyD88) signaling pathway in a rat model of traumatic acute lung injury (ALI). Methods: Thirty SPF healthy male Sprague-Dawley rats, aged 8 weeks, weighing 190-210 g, were divided into 3 groups (n=15 each) by a random number table method: sham operation group (group Sham), traumatic ALI group (group ALI) and group PHCD.ALI was induced by blunt chest trauma in ALI and PHCD groups.PHCD 2 mg/kg was intraperitoneally injected immediately after blunt chest trauma in group PHCD.The rats were sacrificed and lung tissues were removed at 8 h after the model was successfully established for examination of the pathological changes and ultrastructure of lung tissues (with a light microscope or an electron microscope) and for determination of the wet to dry weight ratio (W/D ratio) and expression of TLR4 and MyD88 in lung tissues. Results: Compared with group Sham, the W/D ratio was significantly increased, TIPE2 expression was down-regulated, and the expression of TLR4 and MyD88 was up-regulated in ALI and PHCD groups (P<0.05). Compared with group ALI, the W/D ratio was significantly decreased, TIPE2 expression was up-regulated, and the expression of TLR4 and MyD88 was down-regulated (P<0.05), and the pathological changes of lung tissues and ultrastructure were significantly attenuated in group PHCD. Conclusion The mechanism by which PHCD reduces traumatic AIL is related to activating TIPE2-TLR4-MyD88 signaling pathway in rats.