Objective This research aimed to investigate the expression and mechanism of miR-155 and suppressor of cytokine signaling 1(SOCS1)in rheumatoid arthritis(RA).Methods RT-PCR and flow cytometry were applied to detect the expression differences of miR-155,Th17,and Treg cells in peripheral blood of RA patients(RA group)and control group(HC group).Bioinformatics analysis and dual-luciferase reporter assay were conducted to investigate the regulatory relationship between miR-155 and SOCS1.CD4+T cells were isolated from peripheral blood of RA patients and transfected with miR-155 inhibitor,si-SOCS1,and their respective negative control sequences,and divided into four groups:miR-NC group,miR-155 inhibitor group,miR-155 inhibitor+si-NC group,and miR-155 inhibitor+si-SOCS1 group.The cells were treated with Th17-inducing differentiation medium,and flow cytometry was used to determine the ratio of Th17 cells in each group of CD4+T cells.Western blot was used to determine the ratio of p-STAT3/STAT3 in the cells.Results Compared to the HC group,RA patients showed increased expression of miR-155 and Th17 ratio(P<0.01),and decreased Treg cell ratio(P<0.01).MiR-155 could target and inhibit the expression of SOCS1.Compared to the miR-NC group,the miR-155 inhibitor group,miR-155 inhibitor+si-NC group,and miR-155 inhibitor+si-SOCS1 group showed decreased Th17 ratio and p-STAT3/STAT3 ratio(P<0.01).Compared to the miR-155 inhibitor group,the miR-155 inhibitor+si-SOCS1 group exhibited increased Th17 ratio and p-STAT3/STAT3 ratio in CD4+T cells(P<0.05).Conclusion Elevated expression of miR-155 in RA patients may mediate the differentiation of CD4+T cells into Th17 cells through the SOCS1/STAT3 pathway,contributing to the imbalance of Th17/Treg cells in peripheral blood of RA patients.

Objective:To investigate the effect of low expression of silencing information regulator-6 (SIRT-6) on inflammatory reaction and autophagy in monocytes.Methods:Human acute monocytic leukemia cell line THP-1 was transfected with si-SIRT6 to establish THP-1 cell line with low expressed SIRT-6. The cells were divided into control group, MUS group and MUS+ RAPA group. Cells in control group were cultured with medium added with PBS, cells in MUS Group were cultured with medium added with MUS, and cells in MUS+ RAPA Group were added with MUS and Rapamycin. Cells in each group were cultured for 48 hours. The levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the supernatant of each group were detected by enzyme-linked immunosorbent assay (ELISA). The gene expression levels of autophagy-associated protein-5 (ATG-5), Beclin-1, lysosomal-associated membrane protein-1 (LAMP-1), microtubule-associated protein-1 light chain 3B (LC3B) and p62 in cells of each group were detected by Q-PCR. The protein expression levels of p62, ATG-5 and LC3B Ⅱ/LC3BⅠ in cells of each group. The one-way analysis of variance (ANOVA) was used for the measurement data in multi-groups, and the LSD- t test was used for the measurement data in both groups. Results:The gene and protein expression of SIRT-6 in THP-1 cells decreased significantly after si-SIRT6 transfection (Gene: 1.09±0.08 vs. 0.57±0.03, t=14.91, P<0.001; Protein: 0.21±0.04 vs. 0.12±0.03, t=4.41, P=0.070). The levels of IL-1β, IL-6, and TNF-α in the supernatant of si-SIRT6/si-SIRT6 NC-transfected THP-1 cells increased significantly by MUS ( P<0.05), and the levels of IL-1β, IL-6, and TNF-α in the supernatant of cells further increased by MUS ( P<0.05). The levels of IL-1β, IL-6 and TNF-α in the supernatant of si-SIRT6-transfected THP-1 cells increased significantly compared with those of si-SIRT6 NC-transfected THP-1 cells ( P<0.05). The gene expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly decreased by MUS ( P<0.05), the gene expression of ATG-5, Beclin-1, LAMP-1 and LC3B in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly increased by MUS ( P<0.05). The gene expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further decreased by RAPA ( P<0.05), the gene expression of ATG-5, Beclin-1, LAMP-1 and LC3B in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further increased by RAPA ( P<0.05). The gene expression level of p62 in si-SIRT6 transfected THP-1 cells significantly decreased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05), and the gene expression level of ATG-5, LC3B, Beclin-1 and LAMP-1 significantly increased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05). The protein expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly decreased by MUS ( P<0.05), the protein expression of ATG-5 and LC3B Ⅱ/LC3BⅠ protein in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly increased by MUS ( P<0.05). The protein expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further decreased by RAPA P<0.05), the protein expression of ATG-5 and LC3B Ⅱ/LC3BⅠ in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further increased by RAPA ( P<0.05). The protein expression level of p62 in si-SIRT6 transfected THP-1 cells significantly decreased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05), and the protein expression level of ATG-5 and LC3B Ⅱ/LC3BⅠ significantly increased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05). Conclusion:Low expression of SIRT-6 gene can promote inflammatory reaction and autophagy in monocytes, and Monosodium urate and autophagy agonist rapamycin can aggravate inflammatory reaction and autophagy.

Background: Cardiopulmonary resuscitation (CPR) strategies in COVID-19 patients differ from those in patients suffering from cardiogenic cardiac arrest. During CPR, both healthcare and non-healthcare workers who provide resuscitation are at risk of infection. The Working Group for Expert Consensus on Prevention and Cardiopulmonary Resuscitation for Cardiac Arrest in COVID-19 has developed this Chinese Expert Consensus to guide clinical practice of CPR in COVID-19 patients. Main recommendations: 1) A medical team should be assigned to evaluate severe and critical COVID-19 for early monitoring of cardiac-arrest warning signs. 2) Psychological counseling and treatment are highly recommended, since sympathetic and vagal abnormalities induced by psychological stress from the COVID-19 pandemic can induce cardiac arrest. 3) Healthcare workers should wear personal protective equipment (PPE). 4) Mouth-to-mouth ventilation should be avoided on patients suspected of having or diagnosed with COVID-19. 5) Hands-only chest compression and mechanical chest compression are recommended. 6) Tracheal-intubation procedures should be optimized and tracheal-intubation strategies should be implemented early. 7) CPR should be provided for 20-30 min. 8) Various factors should be taken into consideration such as the interests of patients and family members, ethics, transmission risks, and laws and regulations governing infectious disease control. Changes in management: The following changes or modifications to CPR strategy in COVID-19 patients are proposed: 1) Healthcare workers should wear PPE. 2) Hands-only chest compression and mechanical chest compression can be implemented to reduce or avoid the spread of viruses by aerosols. 3) Both the benefits to patients and the risk of infection should be considered. 4) Hhealthcare workers should be fully aware of and trained in CPR strategies and procedures specifically for patients with COVID-19.

Objective:To explore the clinicopathological characteristics, immunophenotype, diagnosis and differential diagnosis of renal mucinous tubular and spindle cell carcinoma (MTSCC), and to explore the all-exon mutations, microsatellite stability and tumor mutational burden (TMB) in MTSCC cases.Methods:The data of 5 patients with MTSCC that were submitted to the Department of Pathology, First Affiliated Hospital of Soochow University, China from January 2008 to May 2020, were reviewed and analyzed. The whole exome sequencing (WES) was conducted in all patients, while 3 of them were subject to the analyses of microsatellite stability and TMB.Results:Among the 5 patients, 3 were males and 2 were females. They were 37-76 years old. The maximum diameter of the tumor was 3.5-6.0 cm. The borders of the tumors were well defined. Microscopically, MTSCC was characterized by tubular structure, spindle cell and mucinous stroma, and the nuclear grade of tumor cells was overall low. The average follow-up was 15 months, and no recurrence or metastasis was found. Immunohistochemistry showed that all 5 cases were positive for broad-spectrum cytokeratin (CKpan), cytokeratin (CK)7, CK19, vimentin, PAX8, and P504s (varying expression levels), and the Ki-67 positive index was low. The WES of 5 cases showed that NF2 and PTPN14 exhibited higher mutation rates, which were 3/5 and 2/5, respectively. The microsatellite stability analysis indicated that the 3 cases were all microsatellite stable, and the TMB analysis showed that the TMB of the 3 cases were all <9 mut/Mb.Conclusions:MTSCC is a unique, low-grade pleomorphic kidney tumor. The WES analyses suggest that NF2 and PTPN14 have a higher mutation rate, indicating that the occurrence and development of MTSCC may be closely related to the Hippo pathway. The analysis of microsatellite stability indicates that there is no significant relationship between microsatellite stability and MTSCC, and the TMB analysis suggests that MTSCC patients may not benefit from immunotherapy.

The clinical data and the genetic study results of a patient with Liddle syndrome were reported.The genomic DNA of peripheral blood mononuclear cells was extracted and the mutation sites of all the exons in 36 genes related to hypokalemia were screened by high-throughput sequencing.Genetic validation of patients and their parents was performed by direct sequencing.The patient presented with severe hypokalemia,low aldosterone and hypertension.The results of gene sequencing showed that the SCNN1B gene exon 13 584 codon 1751C-T,and the corresponding encoded amino acid changed from alanine to valine (A-V).Eleven families and 33 cases of Liddle syndrome diagnosed by genetic analysis were reported in Chinese literature from 1998 to June 2018.The literature review showed that all patients had hypertension,87％ (28/32) had hypokalemia and 85％ (23/27) had low aldosterone.The onset age of 21 patients was＜20 year,among whom cerebral stroke occurred in 3 patients.The normal blood pressure can be achieved by low-salt diet and administration of amiloride or triamterene.The most common mutations were missense mutations (7/11).Liddle syndrome is a controllable and treatable disease,early detection,diagnosis and treatment can avoid serious complications.

Lesch-Nyhan syndrome(LNS) is a congenital X-linked recessive inherited disorder caused by mutations in the hypoxanthine guanine phosphoribosyl transferase (HPRT) gene.A deficiency of the HPRT enzyme is responsible for the disease.The main clinical manifestation includes hyperuricemia, juvenile-onset gouty arthritis and neurological developmental disorders.Studies have reported there are more than 400 HPRT gene mutation sites, but the incidence of LNS in the Chinese population is extremely low.Here we report a 16-year-old male patient who suffered neurological dysfunction at an early age and gouty arthritis in his youth.DNA of patient and his family members were extracted from peripheral blood lymphocytes.The coding region and the intron-exon boundaries of HPRT gene were sequenced by standard methods.We found a mutation in exon 3 of the HPRT gene of the patient and his mother (Exon3:c.143G>A), which resulted in an arginine to histidine (p.R48H) substitution in the encoded protein.No activity of the enzyme HPRT was detected in the erythrocytes.The same mutation was reported in several European families, but was found in Chinese family for the first time.Clinicians in China have poor experience in diagnosing LNS case, due to the low incidence in China.Therefore LNS screening for infants or adolescents with hyperuricemia, gouty arthritis and neurological dysfunction should be performed.

Objective To observe the effect of rosuvastatin calcium on lipid,vascular endothelial growth factor (VEGF),nitric oxide (NO),tumor necrosis factor (TNF)-α and interleukin (IL)-1 in hyperlipidemia patients.Methods One hundred and twenty-seven hyperlipidemia patients were randomly divided into two groups.Patients in the study group included 72 patients which were given rosuvastatin calcium 10 mg and enteric-coated aspirin 100 mg,orally,once a day for 8 weeks.The control group included 55 patients which were only given enteric-coated aspirin 100 mg,orally,once a day for 8 weeks.The change of lipid,VEGF,NO,TNF-α and IL-1 was observed before and after treatment.Results Before treatment,the level of total cholesterol(TC),triglyceride (TG),low density lipoprotein-cholesterol (LDL-C),high density lipoprotein-cholesterol (HDL-C),VEGF,NO,TNF-α and IL-1 in two groups had no significant difference (P ＞ 0.05).After treatment,the level of TC,TG,LDL-C,TNF-α and IL-1 in study group were significantly lower than those in control group [(4.410 ± 0.688) mmol/L vs.(6.491 ± 0.744) mmol/L,(1.762 ± 0.834) mmol/L vs.(2.632 ± 0.792) mmol/L,(2.256 ± 0.347) mmol/L vs.(4.544 ± 0.493) mmol/L,(41.14 ± 5.41) ng/L vs.(71.34 ± 6.76) ng/L,(0.22 ± 0.18) μ g/L vs.(0.42 ± 0.23) μ g/L] (P ＜ 0.05).The level of HDL-C,VEGF and NO in study group were significantly higer than those in control group [(1.807 ± 0.730) mmol/L vs.(1.432 ± 0.514) mmol/L,(564.86 ± 120.02) ng/L vs.(451.23 ± 100.72) ng/L,(42.39 ± 6.71) μ mol/L vs.(33.65 ± 6.24) μ mol/L](P＜ 0.05).No adverse reaction occurred in two groups.Conclusions Rosuvastatin calcium can obviously decrease the level of lipid,elevate the expression of VEGF and NO,and reduce the expression of TNF-α and IL-1.Rosuvastatin calcium can improve vascular endothelial function obviously in hyperlipidemia patients.

ObjectiveTo explore the diagnosis and prognosis value of clinical pneumonic infection score (CPIS) in patients with stroke-associated pneumonia (SAP).MethodsOne hundred and fifty stroke patients were evaluated and analyzed by CPIS.SAP was regarded as gold standard,and sensitivity,specificity,diagnose accordance rate,positive predictive value and negative predictive value were calculated.Results Thirty-nine patients had SAP and 111 patients did not have SAP by CPIS,and the incidence of SAP was 22.0％ (33/150).CPIS diagnostic sensitivity was 84.8％ (28/33),specificity was 90.6％ ( 106/117 ),positive predictive value was 71.8％ (28/39),negative predictive value was 95.5％ (106/111),and diagnose accordance rate was 89.3％(134/150).The patients of SAP were divided into good prognosis (28 cases) and bad prognosis (5 cases),and the CPIS was significantly lower in patients of good prognosis 7 days after SAP than that in patients of bad prognosis[(4.21 ± 2.23) scores vs. (6.05 ±2.32) scores,P ＜0.05].ConclusionsNot only CPIS has higher diagnosis rate in SAP incidence,but also has good judgment to prognosis.It is worthy of clinical application.

Objective To study the effect of blood glucose level and duration of type 2 diabetes mellitus on carotid intima-media thickness (CIMT) and quantitative carotid stiffness (QCS) in the patients with type 2 diabetes mellitus. Methods Two hundred and seventy-four type 2 diabetic patients (diagnosed according to the World Health Organization criteria) were involved in this study. The subjects were divided into three groups according to the HbAlclevels and duration of diabetes mellitus: 60 with duration ≤2 years (as newly diagnosed) , 96 with duration≥5 years and HbAlc <6.5%, and 118 with duration≥5 years and HbAlc≥6.5%. CIMT and QCS of the common carotid artery were assessed by the high-resolution B-mode ultrasound unit and an echo-tracking system integrated in an uhrasound unit respectively. Results (1) In categorical analyses, the mean level of CIMT was increased from 720 μm in subjects with a duration of diabetes mellitus ≤2 years to 770 μm in subjects with a duration of diabetes mellitus > 2 years and plasma HbAlc< 6.5% (P < 0.05), and to 800 μm in subjects with a duration of diabetes mellitus > 5 years and plasma HbAlc≥ 6.5 % (P < 0.01). The corresponding values for Q CS values were 4.5, 4.5 and 5.1 (P <0.05), respectively. (2) In multiple stepwise regression analyses, CIMT wassignificantly associated with the duration of diabetes mellitus, systolic blood pressure and serum concentration of total cholesterol, whereas QCS was significantly associated with age, duration of diabetes mellitus, smoking, HbAlc, systolic blood pressure (P < 0.05 or P < 0.01). Conclusion QCS is suitable in evaluating the impact of blood glucose on carotid artery, while CIMT is an appropriate tool for long-term follow-up of diabetic patients.

Objective To study the relationship between hyperferritinemia and early metabolism disturbance of sugar and Lipid.Methods 382 people in health examination from Hangzhou First People's Hospital were divided into hyperferritin group(A group)and hypoferritin group(B group)according to the level of ferritin.Blood fat,fasting blood glucose(FBG),uric acid(UA),hematoglobin(Hb),red blood cell count(RBC),hematocrit(HCT),mean corpuscular-hemoglobin concentration(MCHC)were determined,and adiposis hepatica was judged by ultrasonic inspection,then we analyzed by meams of statistics.Results The result showed that TG,LDL,HDL,ApoA1,ApoB,FBG,UA,Hb,RBC,HCT,MCHC had significant deviation between two groups.Ferritin was positive correlated with TG,TC,LDL,ApoB,Hb,RBC,HCT,and negative correlated with HDL,ApoA1.The incidence of adiposis hepatica in A group was obviously higher than B group.Conclusions Probably,ferritin was one of the markers of early metabolism disturbance of sugar and Lipid.