Purpose: To investigate the effect and underlying mechanism of RAR related orphan receptor A (RORA) on preeclampsia (PE). Materials and Methods: Differentially expressed genes (DEGs) in four datasets were obtained by using the Venn diagram method. RORA mRNA and protein expressions were detected by qRT-PCR, western blot, and immunohistochemistry. HTR-8/SVneo cell viability, proliferation, invasion, migration, and angiogenesis were detected by CCK-8 assay, EdU assay, Transwell, wound healing assay, and tube formation assay, respectively. The concentration of Ang-1 in cells was assessed using available ELISA kit.Epithelial-mesenchymal transition, proliferation, and angiogenesis-related proteins were detected by western blot. GSEA analysis were performed for common DEGs, and the expression of enriched pathway-related proteins was also detected. Results: The expression of RORA was increased in PE tissue and HTR-8/SVneo cells. Silencing RORA could promote the migration, invasion, epithelial-mesenchymal transition, proliferation, and angiogenesis of hypoxia-treated HTR-8/SVneo cells. Mechanistically, RORA contributed to the deterioration of PE by activating the JAK2/STAT3 signaling pathway to promote cell proliferation, migration, invasion, and angiogenesis. Conclusion RORA was up-regulated in PE and affected HTR-8/SVneo cell proliferation, invasion, migration, apoptosis, and angiogenesis via the JAK2/STAT3 signaling pathway. This provided a novel strategy for the prevention and treatment of PE.

BACKGROUND: Coronavirus disease 2019 (Covid-19) remains a serious health threat worldwide. We aimed to investigate whether low molecular weight heparin (LMWH) can promote organ function recovery in moderate Covid-19 pneumonia patients. METHODS: We initiated an LMWH protocol in Covid-19 patients with increased D-dimer, body mass index >30 kg/m2 or a history of diabetes from January 18, 2020 at Shanghai Public Health Clinical Center. In this retrospective study, we assigned moderate Covid- 19 pneumonia patients admitted between January 18th and April 18, 2020 receiving the LMWH protocol to the LMWH group. Moderate patients who met the inclusion criteria but did not receive LMWH protocol were included in the control group by 1:2 propensity score matching. General clinical information, indicators for renal function, arterial blood gas analyses, arterial blood lactic acid content (mmol/L), and coagulation indexes at 0 day, 3 days, 7 days, and 11 days after admission were recorded and compared between the two groups. RESULTS: There were 41 patients in the LMWH group and 82 patients in the control group. General information in both groups were similar. Compared to the control group, the arterial blood lactic acid content (mmol/L) at day 11 (1.3 [1.1, 1.7] vs. 1.2 [0.9, 1.3], P = 0.016) was reduced in the LMWH group. The estimated glomerular filtration rate (eGFR) in the LMWH group was higher than that in the control group at day 7 (108.54 [89.11, 128.17] vs. 116.85 [103.39, 133.47], P = 0.039) and day 11 (113.74 [94.49, 126.34] vs. 128.31 [112.75, 144, 12], P  = 0.003). The serum creatinine levels (Scr) in the LMWH group were lower than that in the control group at day 7 (62.13 [51.47, 77.64] vs. 55.49 [49.50, 65.75], P = 0.038) and day 11 (63.35 [50.17, 75.73] vs. 51.62 [44.62, 61.24], P = 0.005). CONCLUSIONS: LMWH treatment can reduce arterial blood lactic acid levels and improve eGFR in moderate Covid-19 pneumonia patients. Randomized controlled trials are warranted to further investigate this issue. TRIAL REGISTRATION ChiCTR.org.cn, ChiCTR2000034796.
COVID-19 ; China ; Glomerular Filtration Rate ; Heparin, Low-Molecular-Weight/therapeutic use* ; Humans ; Lactic Acid ; Retrospective Studies

Objective:To summarize the application of phage therapy in patients with urinary tract complicated pandrug-resistant Klebsiella pneumoniae infection, and analyze its feasibility and effectiveness.Methods:To retrospectively analyze the clinical data of a patient with complicated urinary tract complex pan-resistant Klebsiella pneumoniae treated by phage from August to September, 2019 in Shanghai Public Health Clinical Center. The female patient, 65 years old, was admitted to the hospital on August 6, 2020. The patient repeated with frequent micturition and urgent micturition half a year before admission. These symptoms were not accompanied by back pain, fever, chills, dysuria, gross hematuria. Urinary culture results in outpatient hospital was pan-resistant Klebsiella pneumoniae. After the patient discontinued application of cefoperazone sulbactam, levofloxacin and other drugs, symptoms such as frequent urination could be relieved after treatment, but appeared repeatedly. In August 2019, the center innovatively applied phage therapy to treat this patient with urinary tract pandrug-resistant bacteria infection.Results:For the first time, we applied 117, 135, 178, GD168 phage mixed solution once a day, for 5 days of continuous bladder infusion. At the same time, meropenem and amikacin was intravenous administration to strengthened the anti-infection treatment. Urine culture was negative for two consecutive times after treatment. However, half a month after the end of the bladder infusion, the patient experienced discomfort such as frequent urination. Urine culture: pan-resistant Klebsiella pneumoniae. The second time, we applied a mixture of three phage strains 130, 131, 909, once a day, for 5 days of continuous bladder infusion. And in the afternoon of the third day of treatment, the renal pelvis was retrogradely intubated and perfused with the above three strains of phage mixture. During the second treatment follow-up until March 30, 2020, the patient's urine culture was reviewed once a month. As a result, no pan-resistant Klebsiella pneumoniae was found, and the patient no longer experienced frequent urination and other symptoms of urination. The treatment process was successful and without severe complications and side effects.Conclusions:Phage urinary tract perfusion is an effective method for the treatment of pan-resistant Klebsiella pneumoniae urinary tract infections. The curative effect is accurate and reliable. The patient did not show obvious complications and adverse reactions during treatment. It can be used as an alternative treatment plan for complex pan-resistant Klebsiella pneumoniae infection.

Objective:To explore the in vitro antiviral activity of Mizoribine (Miz) against BK polyomavirus (BKV) and analyze preliminarily the replication stage during inhibition.Methods:The solvent of Miz, was employed as a negative control, while Sirolimus (Sir) with in vitro anti-BKV activity was applied as a positive control. Firstly, the half maximal inhibitory concentrations (IC50) of Miz and Sir were analyzed in different cell lines. Then, prior to BKV infection, cells were treated with a gradient of drug concentrations according to the IC50 results. At different timepoints post-infection, BKV replication curves were measured by quantitative detection of its DNA in supernatant while the cells were subjected to immunofluorescence for detecting the infection rate. Finally 293FT cells treated with drugs was infected by BKV single-round pseudovirus for determining the effects of Miz on the early stage of BKV infection.Results:As compared with control group, immunofluorescence showed a lowered infection rate of BKV in a Miz dose-dependent manner, viral replication curve was significantly inhibited according to the quantitative detection of viral DNA in cell supernatant. The inhibitory effect of Sir on the level of BKV infection and replication was similar to that of Miz. However, neither Miz nor Sir exhibited a significant effect on the early stage of BKV infection.Conclusions:Miz has anti-BKV activity at cellular level and the inhibitory effect does not appear in the early stage of viral infection.

Objective To analyze the clinical features of patients with coronavirus disease 2019 (COVID-19) in Shanghai and to investigate the risk factors for disease progression to severe cases. Methods The clinical data of 292 adult patients with COVID-19 hospitalized in Shanghai Public Health Clinical Center from January 20, 2020 to February 10, 2020 were retrospectively analyzed, including 21 severe patients and 271 mild patients. The demographic characteristics, epidemiological history, history of underlying diseases and laboratory examinations were compared between the two groups. Measurement data were compared using t test or Mann-Whitney U test. The count data were compared using hi-square test. The binary logistic regression equation was used to analyze the risk factors for the progression of patients to severe cases. Results Among the 292 patients, 21 were severe cases with the rate of 7.2% (21/292). One patient died, and the mortality rate was 4.8% in severe patients. The severe patients aged (65.0±15.7) years old, 19 (90.5%) were male, 11 (52.4%) had underlying diseases, 7 (33.3%) had close relatives diagnosed with COVID-19. The mild patients aged (48.7±15.7) years old, 135 (49.8%) were male, 74 (27.3%) had underlying diseases, 36 (13.3%) had close relatives diagnosed with COVID-19. The differences between two groups were all significant statistically ( t =-4.730, χ 2 =12.930, 5.938 and 4.744, respectively, all P <0.05). Compared with the mild patients, the levels of absolute numbers of neutrophils, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, creatinine, serum cystatin C, C reactive protein (CRP), procalcitonin , D -dimer, pro-B-type natriuretic peptide (proBNP), serum myoglobin, creatine kinase (CK), creatine kinase isoenzyme (CK-MB), serum troponin I (cTnI) in severe patients were all significantly higher ( U =2 091.5, 1 928.0, 1 215.5, 729.0, 1 580.5, 1 375.5, 917.5, 789.5, 1 209.0, 1 434.0, 638.0, 964.5, 1 258.0 and 1 747.5, respectively, all P <0.05), while the levels of lymphocyte count, albumin, transferrin, CD3 + T lymphocyte count, CD8 + T lymphocyte count and CD4 + T lymphocyte count in severe patients were all significantly lower ( U =1 263.5, t =4.716, U =1 214.0, 962.0, 1 167.5 and 988.0, respectively, all P <0.05). Further logistic regression analysis showed that the albumin (odds ratio ( OR )=0.806, 95% CI 0.675-0.961), CRP ( OR =1.016, 95% CI 1.000-1.032), serum myoglobin ( OR =1.010, 95% CI 1.004-1.016), CD3 + T lymphocyte count ( OR =0.996, 95% CI 0.991-1.000) and CD8 + T lymphocyte count ( OR =1.006, 95% CI 1.001-1.010) at admission were independent risk factors for the progression of COVID-19 patients to severe illness (all P <0.05). Conclusions Severe cases of patients with COVID-19 in Shanghai are predominantly elderly men with underlying diseases. Albumin, CRP, serum myoglobin, CD3 + T lymphocyte count and CD8 + T lymphocyte count could be used as early warning indicators for severe cases, which deserve more clinical attention.

Objective To explore the protective effect of ETaR siRNA on renal ischemia reperfusion injury (IRI) by changing the immuno-microenvironment in rats .Methods A total of 40 male Sprague-Dawley (SD) rats were randomized into four groups of sham ,IR ,negative siRNA and ETaR siRNA .A renal IRI model was generated by clamping left renal artery .ETaR siRNA was delivered into kidney through renal vein by a retrograde 'hydrodynamic' injection .Blood samples were collected for detecting renal function and kidney tissue harvested for Hematoxylin & Eosin (HE) staining , TdT-mediated dUTP Nick-End Labeling (TUNEL) staining ,polymerase chain reaction (PCR) and Western blot at 48 h post-reperfusion .Results Serum creatinine ,blood urea nitrogen and renal apoptotic cells increased and renal tissue was injured after IR . The changes were inhibited by ETaR siRNA . PCR showed that ETaR siRNA treatment significantly down-regulated the expressions of inflammatory factors TNF-α , IFN-γ and IL-6 and transcription factor NF-κB induced by IR .Conclusions ETaR siRNA can effectively improve the immunomicroenvironment and thereby alleviate renal ischemia reperfusion injury .

Objective To explore the efficacy of renal transplantation plus hematopoietic stem cell transplantation on inducing immune tolerance and summarize its long-term follow-up outcomes . Methods From 2009 to 2018 ,a total of 11 cases of living related donor kidney transplantation plus hematopoietic stem cell transplantation were performed .Two of them were HLA-matched and the remainder were mismatched for one HLA haplotype . The donor hematopoietic stem cells were mobilized using granulocyte colony-stimulating factor at 5 days pre-transplantation and collected at 1 day pre-operation .The recipients received total lymphoid irradiation for 3 days pre-transplantation and received anti-thymocyte globulin induction during transplantation .The donor hematopoietic stem cells were infused at 2 ,4 and 6 postoperative day .Postoperative regulatory T cells ,chimerism ,B cell activating factor and mixed lymphocyte culture and other parameters were detected and long-term follow-up outcomes tracked .Results The immune tolerance-inducible recipients had a significant increase in activated Treg .One HLA-matched recipient achieved 30％-50％ of chimerism and lost after 6 months .However ,other recipients did not achieve mixed chimerism .The BAFF of recipient spiked sharply after transplantation .Mixed lymphocyte culture indicated that a donor-specific low response was induced .The recipients were followed up for 717 to 3612 days .The first recipient lost renal function and another ten recipients had stable renal function . None of the recipients had myelosuppression or graft-versus-host disease .Allograft biopsy confirmed only one case of mild acute rejection . The dose of immunosuppressive agents was lowered in 5 patients .Conclusions Hematopoietic stem cell transplantation for inducing tolerance is safe during renal transplantation . And chimerism is essential for inducing immune tolerance .

Objective To evaluate the efficacy and safety of single bolus high dose (SD group) ATG-Fresenius induction therapy in kidney transplantation vs.multiple low dose (MD group) administration.Methods A multiple center,prospective,randomized and controlled clinical study was performed on 280 de novo renal transplant recipients from 19 centers.Patients were randomized into 2 groups as follows:SD group,a single high dose (7-9 mg/kg) of ATG-F infused as an induction agent before the vessel anastomoses;MD group,2 mg/kg of ATG-F daily administrated in postoperative 4 days.All the patients accepted maintenance immunosuppressive protocol including tacrolimus,mycophenolate and prednisone.Patients were assessed and data were collected at regular schedule clinic visits on the day 1,3,7,14,30,90,180,270 and 365.The primary end point of efficacy was therapeutic failure rate [the number of death,grafts loss and acute rejection (AR)].The event first occurred should be used in the classification of patients.The non-inferiority evaluation of the two treatment regimens was done based on treatment failure rate.The secondary end points of efficacy were the incidence of AR,delayed graft function (DGF),1-year survival rate of patients and grafts,and serum creatinine at each visiting point.The indicators for safety evaluation included hemotologic variation and incidence of adverse events.Results The therapeutic failure rate in SD group was non-inferior to the MD group (17.24％ vs.23.08％).AR was the major cause of therapeutic failure and there was similar incidence of AR between SD gronp and MD group (12.07％ vs.21.37％).There was no significant difference in the incidence of DGF between SD group and MD group (12.07％ vs.6.84％,P =0.1721).The 1-year patient's survival rate and 1-year graft survival rate in SD group and MD group showed no significant difference (96.55％ vs.98.29％,P =0.6714;94.83％ vs 98.29％,P =0.2750).The serum creatinine level showed no significant differences between two groups at each visit point.There was also no significant difference in total incidence of adverse events between the two groups.In addition,there was also no statistically significant difference in the incidence of concerned and drug-related adverse events between the two groups,including infection,hemotologic abnormality,liver or renal dysfunction,gastrointestinal disorder,etc.After ATG--F administration,peripheral blood lymphocytes in the SD and the MD group immediately decreased but nearly restored to the normal level on the postoperative day 30 and 90 respectively.No severe granulocytopenia,erythropenia or thrombocytopenia occurred in both two groups.Conclusion The efficacy and safety of single high dose of ATG-F induction are non-inferior to multiple low dose ATG-F induction,moreover,single high dose of ATG-F induction is administered more conveniently and economically.

Objective To explore effect of diosmin on serum and prostatic fluid immune mediated factors and clinical effect in patients with chronic prostatitis.Methods 89 cases of old male patients with chronic non bacterial prostatitis were selected, and divided into two groups.The control group were treated with universal tablets, the experiment group were treated on the base of the control group with diosmin.12 weeks as a course.Clinic effect, adverse reaction rate, prostatic fluid MIP-2 and MIP-1αwere compared after treatment.Results Compared with control group, MIP-1αand MIP-2 in serum and prostatic fluid were lower(P<0.05),NIH-CPSI score and QOL scores were lower(P<0.05), total effective rate was higher than control group(P<0.05),the incidence of adverse reactions was lower(P<0.05).Conclusion Diosmin can reduce MIP-1α, MIP-2 in serum and prostate fluid of patients with chronic none bacterial prostatitis, and improve the pelvic pain, dysuria, sexual dysfunction and other symptoms, and has less adverse reaction.

BACKGROUND:Studies have shown that polyethylene polymer-synthesized ureteral stents have good biocompatibility in vivo, but the side effects are found to induce the body to produce inflammatory, pyrogenic, and al ergic reactions. Therefore, studies have mainly aimed to explore the biocompatibility, toxic and side effects, inflammation of newly developed ureteral stents synthesized by L-lactic acid, glycolide, and barium sulfate. OBJECTIVE:Based on the biocompatibility comparisons on cel toxicity test, inflammatory reaction, al ergic reaction, pyrogen reaction and mortality after implantation between the usage of polyethylene material and L-lactic acid, glycolide, barium sulphate syntheses, to provide a better reference to introduce a new clinical ureteral stent material. METHODS:There were three groups in the experiment:blank control group, new material group, polyethylene group. Cel s were cultured in normal medium, leach liquid of L-lactic acid, glycolide, barium sulphate syntheses and leach liquid of polyethylene, respectively. RESULTS AND CONCLUSION:The toxic effects were significantly enhanced in the urethral epithelial cel s that grew in the leach liquids of two materials compared with the blank control group (P<0.05), but the two kinds of materials had no significant statistical difference in the toxicity test (P>0.05). Hematoxylin-eosin staining results showed that there were obvious inflammatory infiltrations in muscle tissue and the number of neutrophils and eosinophils were increased when the two kinds of materials were implanted in the rats for 2 and 6 weeks (P<0.05), but the inflammatory infiltration and tissue damage in the new material group was less than that in the polyethylene group (P<0.05). ELISA results showed that the serum concentrations of interleukin-10 and interleukin-23 were significantly increased when the two kinds of materials were implanted in the rats for 6 weeks (P<0.05), but the content of inflammatory cytokines in the new material group was less than that in the polyethylene group (P<0.05). In the new material group, there was only one rat that developed al ergic reaction and there was no pyrogen reaction or death;however, in the polyethylene group, there were three rats with presence of al ergic reaction and one rat with pyrogen reaction but no death. These findings indicate that the new ureteral stent material synthesized with L-lactic acid, glycolide and barium sulphate can reduce inflammation reactions, al ergic reactions and pyrogen reactions to the body, and has better biocompatibility than polyethylene composite material.