Objective To define which hyaluronan synthase (HAS), of three hyaluronan synthesizing enzymes HAS-1, HAS-2, and HAS-3, is primarily responsible for hyaluronan synthesis and extracellular matrix/extracellular coat formation in human peritoneal mesothelial cells (HPMCs) . Methods As a prerequisite study, the expression of each HAS mRNA in cultured HPMCs was measured by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). Only the expression of HAS-2 and HAS-3 mRNA could be detected. The level of HAS-2 mRNA expression was about 10 fold higher than that of HAS-3. HAS-2 specific antisense oligonucleotide was then transfected into cultured HPMCs by lipofectamine. After 0, 8, 24, and 48 hours, the expression of HAS-2 mRNA was detected by RT-PCR and the extracellular coat was measured by particle exclusion test. Results 8 hours and 24 hours after transfection, the expression of HAS-2 mRNA in HPMCs decreased by 58% and 89% respectively; 48 hours after transfection, the expression of HAS-2 mRNA in HPMCs partially restored to 25% of the normal level. Correspondingly, 24 hours after transfection, the extracellular matrix/extracellular coat in HPMCs almost completely disappeared. However, as control, sense and reverse oligonucleotides showed no effect. Conclusion HAS-2 plays a leading role in HPMCs hyaluronan synthesis and the formation of extracellualr matrix/extracellular coat.

Objective To study the clinical curative effect and complication of methylprednisolone in the treatment of patients with acute myelitis.Methods 42 patients with acute myelitis were selected and randomly divid-ed into two groups,21 cases in each group.The control group was treated with dexamethasone,the observation group was treated with methylprednisolone.After treatment,the clinical parameters and effect of the two groups were com-pared.Results The total effective rate of the observation group was 85.7%,which was higher than 42.9% of the control group (χ2 =2.41,P <0.05).The improvement of muscle strength,restore micturition,walk on their own time in the observation group were (10.1 ±4.1)d,(7.1 ±3.4)d,(16.1 ±4.2)d,respectively,which were better than (23.4 ±7.3)d,(10.2 ±3.1)d,(22.6 ±7.3)d in the control group(t =3.15,1.69,1.97,all P <0.05).The total incidence of gastrointestinal reaction and complication rates of the observation group was 4.8%,which was lower than 19.0% of the control group (χ2 =6.37,P <0.05).Conclusion In treatment of patients with acute myelitis,methyl-prednisolone has good effect,which should be used in clinical practice.

Aim To investigate whether there is IL-12 signaling pathway through lck/P38/c-jun in splenic cells obtained from lupus mouse and its effect on splenic cells. Methods Mice with graft versus host disease were used as lupus nephritis model. Activity of Lck tyrosine kinase, p38 phosphorylation and mRNA expression of c-jun in splenic cells were determined by autoradiography, Western blot and Northern blot, respectively. Results There were higher levels of Lck activity, p38 phosphorylation and c-jun expression of IL-12-stimulated splenic cells from lupus model when compared with that observed in similarly treated splenic cells from normal control. The Lck activity and p38 phosphorylation were almost inhibited by Lck inhibitor PP1, on the other hand, p35 specific inhibitor SB203580 decreased phosphorylation of p38. In addition, expression of c-Jun was also inhibited by PP1 or SB203580 although splenic cells were stimulated with IL-12. Conclusion Aberrant murine splenic cell, IL-12-me-diated intracelluar signaling pathway through lck/p38/c-Jun were involved in immunologic damage.

AIM: To investigate the effect of antisense RNA on osteopontin (OPN) expression in renal tubular epithelial cells. METHODS: Cell clone expressing stably OPN antisense RNA was formed by transfering retroviral vector expressing OPN antisense RNA into renal tubular epithelial cells, NRK52E cells, using liposome, with cell clones transfected by empty vector and vector expressing OPN sense RNA as controls. Ribonuclease protection assay(RPA), Western Blot, ELISA and assay of OPN activity were performed to detect expression of OPN mRNA and protein in above clones cultured with or without epidermal growth factor(EGF). RESULTS: The antisense RNA was only expressed by antisense clone. Antisense clone, sense clone and empty clone all expressed OPN mRNA. EGF enhanced expression of OPN mRNA, but not OPN antisense RNA or OPN sense RNA in above clones. OPN protein was not expressed in antisense clone cultured with or without EGF and empty clone cultured without EGF, but was expressed in sense clone cultured with or without EGF and empty clone cultured with EGF. CONCLUSION: Antisense RNA can inhibit OPN protein expression by means of preventing OPN mRNA translation, but not inhibit OPN mRNA transcription in renal tubular epithelial cells.

Objective: To study the epidemiology and the etiology characteristics of first imported severe fever with thrombocytopenia syndrome (SFTS) case reported in Shenzhen city in 2017. Methods: Data on descriptive epidemiology was collected to study the characteristics to the epidemic. The serum sample collected from the suspect SFTS case was detected for IgM, IgG by ELISA and severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) nucleic acid by real-time RT-PCR. The samples were further inoculated in Vero cell for virus isolation. The partial fragements of L and S gene were amplified by RT-PCR and sequenced to construct homology comparison and phylogenetic tree with the strains isolated from other areas. Results: The case was laboratory confirmed imported SFTS case in Shenzhen on May 2017. IgM antibody and RNA of SFTSV were detected in the serum sample. SFTSV named GDSZ01/2017/China was successfully isolated from the serum sample. The high nucleotide homology of L and S genome segments were found at 95.3%-98.2% and 93.8%-98.8% with other representative strains from the popular provinces, respectively. The phylogenetic tree indicated that GDSZ01 was most close to SDTA_3 strain, next to strains in Hubei procince. The isolated SFTSV belonged to genotype C3 with HB29, HB154. Conclusions The virological, serological and molecular features showed that the imported case of SFTS in 2017 was caused by SFTSV C3 genotype.

Objective:To verify the clinical value of the good outcome following attempted resuscitation (GO-FAR) score in predicting the neurological status of patients with in-hospital cardiac arrest (IHCA) in the Chinese population.Methods:The clinical data of patients with IHCA who were admitted to the Zigong Fourth People's Hospital from January 1 to December 31, 2020 were retrospectively analyzed. Used Glasgow-Pittsburgh cerebral performance category (CPC) score 1 point as the end point, the subjects were divided into 4 groups according to the score: ≤ 0 group, 1-8 group, 9-20 group and ≥ 21 group. Taken the group which GO-FAR score ≤ 0 as the reference group, the odds ratio ( OR) of the other three groups compared with this group was calculated. The receiver operator characteristic curve (ROC curve) was performed to evaluate the predictive value of the GO-FAR score in favorable neurological outcome. A calibration curve was drawn for the Hosmer-Lemeshow test to analyze the degree of calibration of the GO-FAR score for predicting good neurological outcome. Results:A total of 230 IHCA patients were enrolled in the study, including 130 males, aged 74 (65, 81) years old, and 23 case (10.0%) had good neurological prognosis. There were statistically significant differences in GO-FAR-related variables, including age, a normal neurological function on admitted, acute stroke, metastatic cancer, septicemia, medical noncardiac admission, hepatic insufficiency, hypotension, renal insufficiency or dialysis, respiratory insufficiency, pneumonia, etc (all P < 0.05). Taken the GO-FAR score ≤ 0 group as the reference group, the OR values of good neurological prognosis in the GO-FAR score 1-8 group were 0.54 [95% confidence interval (95% CI) was 0.17-1.53, P = 0.250], 9-20 group were 0.17 (95% CI was 0.02-0.67, P = 0.009) and ≥ 21 group were 0.25 (95% CI was 0.05-0.85, P = 0.025). The area under the ROC curve (AUC) of the GO-FAR score for predicting favorable neurological outcome in IHCA patients was 0.653 (95% CI was 0.529-0.777, P = 0.015) and there was no significant difference in Hosmer-Lemeshow test ( P = 0.311). All these suggested that there was no significant difference between the predicted value and the actual value. Conclusions:GO-FAR score can be applied to predict neurological prognosis of IHCA patients in Chinese population. It can help clinicians to predict the prognosis of cardio-pulmonary resuscitation (CPR) and propose critical recommendations in treatment for these patients or their families.

OBJECTIVETo determine the in vitro expression of interleukin-12 (IL-12) and its effect on signal transducers and activators of transcription (STAT) signaling molecules in peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE).

METHODSPeripheral blood mononuclear cells in 39 patients with definite systemic lupus erythematosus and 11 healthy volunteers were collected. Expression of IL-12 P40mRNA in PBMCs was determined with reverse transcription-polymerase chain reaction (RT-PCR). Quantity of IL-12 protein supernatant was measured by enzyme-linked immunosorbent assay (ELISA). The levels of phosphorylated STAT3 and STAT4 signaling molecules in PBMCs were detected by immunoblot.

RESULTSLevels of IL-12 protein and mRNA expression in patients with active or inactive SLE were significantly higher than those in controls. Phytohemagglutinin (PHA ) may promote the expression of IL-12. IL-12 alone induced the phosphorylation of STAT3 and STAT4 in PBMCs from patients with SLE, especially in active SLE. However it had no obvious effect on normal PBMCs. Phosphorylated STAT3 and STAT4 might be observed in normal PBMCs treated with IL-12 plus PHA.

CONCLUSIONIL-12 is produced aberrantly in patients with SLE. IL-12 might exert its biological role in SLE via the aberrantly phosphorylated STAT3 and STAT4 signaling molecules.

Adolescent ; Adult ; Cells, Cultured ; DNA-Binding Proteins ; metabolism ; Humans ; Interleukin-12 ; blood ; genetics ; Leukocytes, Mononuclear ; metabolism ; Lupus Erythematosus, Systemic ; metabolism ; Middle Aged ; Phosphorylation ; RNA, Messenger ; analysis ; STAT3 Transcription Factor ; STAT4 Transcription Factor ; Trans-Activators ; metabolism

Objective:To investigate the epidemiological characteristics of dengue fever and genotyping of the epidemic strains of dengue virus in Shenzhen City in 2018.Methods:Descriptive epidemiological analysis was used to analyze dengue fever prevalence in Shenzhen City in 2018. Blood samples of patients with dengue fever were collected. The colloidal gold immunochromatography was used to detect serum specific IgM and IgG antibodies, and real-time fluorescence quantitative polymerase chain reaction (PCR) was used to detect viral nucleic acids and to identify genotypes. The E gene sequence of isolated virus strain was amplified by reverse transcription PCR. Homology comparison and phylogenetic tree of dengue fever epidemic strains in different countries and regions were conducted. Results:A total of 234 cases of dengue fever were reported in Shenzhen City from January 1 to December 31, 2018. The incidence rate was 1.87/100 000. There were 144 (61.54%) local patients and 90 (38.46%) imported patients, who were mainly from Southeast Asia and surrounding cities. Two hundred and two (86.32%) cases were reported during the epidemic peak period from August to November of the year. The patients mainly aged 20 to 50 years old (195 cases, 83.33%). Dengue virus type (DENV)-1 accounted for 86.01%(166/193), DENV-2 accounted for 10.36%(20/193), DENV-3 accounted for 2.59%(5/193), and DENV-4 accounted for 1.04%(2/193). The local cases were all infected with DENV-1. The homologies of nucleotide sequence and the deduced amino acid sequence of E gene of 24 DENV-1 strains with HAWAII45 strain were 93.0% to 94.6% and 96.6% to 97.2%, respectively. The phylogenetic tree of DENV-1 strains revealed that 23 strains belonged to genotypeⅠ, and one strain belonged to genotype Ⅳ which was the first reported imported cases in Shenzhen City. The homologies of nucleotide sequence and the deduced amino acid sequence of E gene of six DENV-2 strains with NGC strain were 93.1%to 93.9% and 97.0% to 97.8%, respectively. The phylogenetic tree of DENV-2 strains showed that two strains belonged to genotype Cosmopolitan and four strains belonged to genotype Asian Ⅰ, which were first reported in Shenzhen City. Conclusions:The epidemic of dengue fever in Shenzhen City in 2018 has the characteristics of coexistence of local and imported transmission. The main epidemic genotype is DENV-1. It infers that the major virus strains may be imported from Southeast Asia countries and surrounding cities. Therefore, attention should be paid to the epidemic trend of local dengue fever.

Objective To investigate the clinical efficacy and safety of high-dose caspofungin (70 mg/d)as initial or salvage treatment for invasive pulmonary aspergillosis.Methods Twenty-one patients with proven or probable invasive pulmonary aspergillosis from June 2014 to October 2017 in Huashan Hospital,Fudan University were retrospectively reviewed.According to the anti-fungal treatment before high-dose caspofungin application,patients were divided into initial treatment group and salvage treatment group.Patients' clinical data and laboratory data were collected.The characteristics,clinical efficacy,adverse reactions,one-year survival rate and the overall effective rate were evaluated.The prognosis of the two groups was compared by Kaplan-Meier analysis.Results Twenty of the 21 patients opportunistic acquired invasive pulmonary aspergillosis during the treatment of underlying diseases.Five patients were initially treated with high-dose caspofungin for 68 (62) days.At week 12,one patient achieved complete response,3 patients achieved partial response,and the overall effective rate was 80％ (4/5).Sixteen patients received caspofungin as salvage therapy for 66.50 (58) days,of which one patient got complete response at week 12,10 had partial response,and the overall effective rate was 68.75％ (11/16).One-year follow-up showed that no patient died in the initial treatment group,and the one-year survival rate was 100％ (5/5).In salvage treatment group,3 patients died of pulmonary bacterial infections and the one-year survival rate was 81.25％ (13/16).During treatment,one patient had elevated total bilirubin,which was possibly associated with high-dose caspofungin.Conclusions High-dose caspofungin regimen has good efficacy and safety,both for initial treatment and salvage therapy in patients with invasive pulmonary aspergillosis.

Objective:To investigate the clinical characteristics and prognosis of cryptococcal meningitis patients with anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) autoantibodies.Methods:A total of 216 non-acquired immunodeficiency syndrome (AIDS) related cryptococcal meningitis cases with positive cultures of Cryptococcus, hospitalized at Huashan Hospital, Fudan University during January 2014 and December 2021, were retrospectively included. The serum anti-GM-CSF autoantibodies were detected by enzyme linked immunosorbent assay, and the clinical characteristics and prognosis were compared between patients with and without anti-GM-CSF autoantibodies. Statistical comparisons were mainly performed using the chi-square test or Fisher′s exact test. Cox proportional-hazards model was used to analyze the risk factors associated with prognosis. Results:Among 216 enrolled patients, 23 patients were positive of anti-GM-CSF autoantibodies, with a positive rate of 10.6%. Among 23 patients, seven cases were infected with Cryptococcus gattii, and 16 cases were infected with Cryptococcus neoformans. In the group with positive anti-GM-CSF autoantibodies, 30.4%(7/23) of the patients were infected with Cryptococcus gattii, which was higher than that of 1.6%(3/193) in the group with negative anti-GM-CSF autoantibodies, and the difference was statistically significant ( χ2=38.82, P<0.001). In the group with positive anti-GM-CSF autoantibodies, 30.0% (6/20) had mass lesions with a diameter greater than three centimeters in the lungs, and the one-year all-cause mortality rate was 50.0% (10/20), which were both higher than those of 3.4%(5/145) and 16.1% (29/180) in the negative group, respectively. The differences were both statistically significant (both Fisher′s exact test, P<0.01). Age≥60 years (hazard ratio ( HR)=4.146, P=0.002), predisposing factors ( HR=3.160, P=0.021), epilepsy ( HR=6.129, P=0.002), positive anti-GM-CSF autoantibodies ( HR=2.675, P=0.034), white blood cell count of cerebrospinal fluid (CSF)<100 ×10 6/L ( HR=2.736, P=0.039), the titers of cryptococcal capsular polysaccharide antigen of CSF≥1∶1 280 ( HR=4.361, P=0.009) were independent risk factors for one-year all-cause mortality in patients with cryptococcal meningitis. Conclusions:In non-AIDS related cryptococcal meningitis patients, the positive rate of serum anti-GM-CSF autoantibodies is as high as 10.6%. Patients with anti-GM-CSF autoantibodies could be infected with both Cryptococcus neoformans and Cryptococcus gattii, and they have higher proportion of lung mass lesions than patients with negative anti-GM-CSF autoantibodies. The one-year survival rate decreases significantly in patients with anti-GM-CSF autoantibodies, which is an independent risk factor for the prognosis of cryptococcal meningitis.