Objective To explore whether the use of purified rabbit serum paraoxonase 1 (PON1) for the treatment of dichlorvos-induced liver injury in rats is superior to traditional method.Methods Thirty male SD rats were randomly divided into the followint 5 groups,with 6 rats in each group:control group (A group),dichlorvos group (B group),traditional treatment group (C group),PON 1 treatment group (D group),combined treatment group (E group).Rats in groups B,C,D and E were adminstered dichlorvos by intraperitoneal injection 9 mg/kg.In group C,atropine 10 mg/kg and iodine solution 45 mg/kg were injected intraperitoneally within 2 min after dichlorvos administration.In group D,PON1 was injected intravenously at a dose of 9 600 U/kg,30 min prior to poisoning.In group E,PON1 was injected intravenously at a dose of 9 600 U/kg,30 min prior to poisoning,followed by in travenous injection of atropine 10 mg/kg and iodine solution 45 rng/kg within 2 min after poisoning.Rats in A group received normal saline.Blood was collected at different time points to examine the acetyl cholinesterase (AChE)-levels by ELISA method.Liver tissue were collected at 12 hours after model establishment to observe the pathological changes.The expression of 4 hydroxy 2-nonenal (4-HNE) in the liver tissue was detected by immunohistochemistry and Western blotting.Results In group B,AChE levels decreased significantly,liver cells showed severn fatty degeneration,karyopyknosis and other pathological changes,and 4-HNE expression increased.The pathological changes of group D and group E were less obvious than those of group C,and the 4-HNE expression in the group D and group E were significantly different from that in the group C (P＜ 0.05).Conclusion PON1 plays a protective role in dichlorvos-induced liver injury in rats,and this protection is better than that offered by traditional treatment.

Objective To assess the protective effect of rabbit serum paraoxonase-1 (PON-1) on renal injury induced by dichlorvos in rats.Methods Totally 30 healthy S-D rats were randomly divided into 5 groups:control group (group A,n =6),exposure group (group B,n =6),PON-1 pretreatment group (group C,n =6),traditional atropine,pralidoxime treatment group (group D,n =6) and combination therapy group (group E,n =6).The rats of group A were given normal saline in equal volume of dichlorvos injected into abdominal cavity to make a false model of dichlorvos poisoning.In rats of groups B,C,D and E,9 mg/kg dichlorvos was administered.In rats of groups C and E,PON-1 4 500 units/kg was injected into vein of the tails half an hour before dichlorvos administration.After dichlorvos exposure,rats in group D and E were treated with 45 mg/kg iodoprofen and 10 mg/kg atropine by intraperitoneal injection.The activity of blood urea nitrogen (BUN) was assayed with urease.Serum creatinine (Cr) were measured by picric acid colorimetry.Serum Cys-C,KIM-1 and NAG in urine were determined by ELISA.Ultrastructural changes in renal tissues of rats were examined by light microscopy.The differences in laboratory findings between groups were compared.Results The creatinine level in group B was significantly higher than that in other groups (P ＜0.05).The levels of Cys-C,KIM-1 and NAG in group B and group D were significantly higher than those in group A (P ＜ 0.01).But there were no significant differences in above biomarkers among group C,group E and group A.There were no significant differences in above biomarkers between group B and group D.In group B,inflammatory cells infiltrated extensively in renal tissues and,the renal cells were congested and edematous,the lumen was obliterated and the border of the brush disappeared.The tubular structures were not clearly distinct found in group B,but edema and inflammatory cell infiltration with lesser degree were found in group D than those in dichlorvos exposure groups.The clearly distinct structure of the tube without completely occluded lumen in group D,and the most serious lesions were found in distal convoluted tubules.In group C,and group E,there were only mild congestion and edema without significant cell degeneration and necrosis.In group A,the structure of renal tubular epithelium was clearly distinct with brush-shaped margin,and without tubular or necrotic cell debris in the lumen.Conclusion The rabbit serum PON-1 can protect the renal tissue of rats after dichlorvos exposure.

Objective To discuss the prognosis and its risk factors in patients with acute decompensatory pulmonary hypertension and provide a basis for their clinical management.Methods A total of 287 patients with acute decompensated pulmonary hypertension patients were selected in our hospital's Emergency Intensive Care Unit between April 2015 and April 2020.Patients were divided into a survival group(n=140)and a death group(n=147)according to in-hospital mortality status.The clinical characteristics,laboratory examination results,treatment plans,clinical events,and prognoses were compared between groups.The correlation between the in-hospital mortality rate,Euro-pean Society of Cardiology/European Respiratory Society(ESC/ERS)risk assessment system score,and Sequential Organ Failure Assessment(SOFA)score were evaluated using multivariate regression analysis.The ESC/ERS risk assessment system and SOFA scores were analyzed to evaluate the efficiency of clinical prognosis using a receiver operating characteristic(ROC)curve.Results Compared with the survival group,in the death group,the rates of intermediate and high risk in the ESC/ERS assessment and NYHAⅢ/Ⅳclass were increased(P<0.05),the mean pulmonary artery pressure and N-terminal pro-BNP(NT-proBNP)level were considerably elevated(P<0.05),and the levels of arte-rial partial pressure of oxygen(PaO2)and tricuspid annular plane systolic extrusion were considerably reduced(P<0.05).On a multivariate Cox regression analysis,a SOFA score≥8,high ESC/ERS risk assessment score,and high NT-proBNP and PaO2 levels were independent risk factors for in-hospital mortality(all P<0.05).A ROC curve analysis showed that the area under the curve of the ESC/ERS risk assessment combined with SOFA score was the largest at 0.877(P<0.001),with a sensitivity of 92.5%and specificity of 83.0%.Conclusion High risk on an ESC/ERS risk assessment and a SOFA score≥8 are independent risk factors of in-hospital mortality among patients with acute decom-pensatory pulmonary hypertension.The ESC/ERS risk assessment and SOFA score can be used to evaluate the prognosis of patients with acute decompensated pulmonary hypertension in the Intensive Care Unit;moreover,their combined use has better predictive efficacy.

[Objective]To explore the medication rules of Professor HONG Min,a famous traditional Chinese medicine practitioner in Guangdong Province,for treating insomnia.[Methods]The prescriptions of Professor HONG Min for treating insomnia in clinical practice were collected and sorted out,and data mining and analysis were conducted on the frequency of medication,four properties,five flavors,meridian tropism,prescription patterns,potential new formulas and so on by using the Traditional Chinese Medicine Inheritance Support System Platform.[Results]A total of 509 formulas and 180 Chinese herbs were screened out.The top five Chinese herbs in terms of frequency of use were Semen Ziziphi Spinosae Preparata,Rhizoma Pinelliae Preparatum,Radix Glycyrrhizae Preparata,Poria cocos and Radix Scutellariae.The high-frequency herb combinations included Rhizoma Pinelliae Preparatum-Semen Ziziphi Spinosae Preparata,Radix Glycyrrhizae Preparata-Rhizoma Pinelliae Preparatum,Rhizoma Pinelliae Preparatum-Poria cocos,Radix Glycyrrhizae Preparata-Semen ziziphi spinosae Preparata,Poria cocos-Semen ziziphi spinosae Preparata,Rhizoma Pinelliae Preparatum-Radix Scutellariae,Citri Reticulatae Pericarpium-Poria cocos and so on.Seven potential new formula combinations were also obtained.[Conclusion]Professor HONG Min's treatment of insomnia is mainly based on soothing the liver and calming the mind,clearing heat and drying dampness,strengthening the spleen and resolving phlegm.In clinical practice,she pays attention to the theory of five movements and six Qi combined with the theory of six meridians differentiation.She is good at flexibly applying the theory of Zang fu tonification and purgation,balancing multiple organs simultaneously.She also pays attention to integrating constitution,movement of Qi,pathogenesis,human nature,heaven nature and disease nature,comprehensively regulating and treating them to achieve harmony between Yin and Yang and curing insomnia.

Objective To observe the effects of three treatment methods on renal anemia in maintenance hemodialysis patients with hyperparathyroidism secondary to uremia and analyze the influencing factors of erythropoietin (EPO) dosage. Methods A total of 55 maintenance hemodialysis patients with secondary hyperparathyroidism at the hemodialysis center of Huashan Hospital affiliated to Fudan University from January 2015 to December 2016 were retrospectively divided into three groups according to different treatment methods, parathyroidectomy +forearm transplantation group (surgery group, n=16), cinacalcet treatment group (n=6), and calcitriol treatment group (n=33), respectively. The hemoglobin level and erythropoietin dosage were measured before treatment and in the 3rd month, the 6th month and the 12th month after treatment. The changes of hemoglobin and erythropoietin dosage in the three groups before and after treatment were observed, and the mixed effect model was used to analyze the difference of the change of hemoglobin and erythropoietin dosage among three groups. Multiple linear regression analysis was used to analyze the influencing factors of EPO dosage after one year. Results The levels of intact parathyroid hormone (iPTH) in the surgery group and the cinacalcet group before treatment were significantly higher than that in the calcitriol group (both P<0.05). In the 12th month after treatment, the levels of iPTH decreased significantly in the patients of surgery group and the cinacalcet group compared with those before treatment (both P<0.05). The levels of serum alkaline phosphatase, serum calcium and serum phosphorus in the surgery group also decreased significantly compared with those before treatment (all P<0.05). The mixed effect model analysis showed that the hemoglobin level of surgery group was on an upward trend after the treatment, and the overall level was significantly higher than cinacalcet and calcitriol treatment group (P=0.007). There was no significant difference in the dosage change of erythropoietin (EPO) in the three groups over time. However, the intra-group comparison of the mixed effect model showed that the dosage of EPO in the 12th month was significantly lower than that of before the treatment in surgery group (P=0.007). Multiple linear regression analysis showed that dialysis vintage (B=-0.064, P=0.012) and ferritin ≥ 500 μg/L (B=0.645, P=0.032) were independent influencing factors of EPO dosage. The longer the dialysis vintage, the less EPO dosage, and more EPO dosage were observed in patients with ferritin ≥ 500 μg/L. Conclusions Parathyroidectomy and forearm transplantation is more effective in reducing EPO dosage and improving renal anemia in maintenance hemodialysis patients with secondary hyperparathyroidism. Dialysis vintage and ferritin are independent influencing factors for the dosage of EPO.

Objective: To investigate the inactivating effect of soap solution on rabies virus and to explore the concentration of soap solutions which could be effective in post-exposure prophylaxis (PEP) of rabies virus infection. Methods: The BSR and N2a cells were respectively infected by the mixture of different concentrations of soap solution and rabies virus CVS-11. Based on the direct immunofluorescent method (DFA) and reverse transcription PCR (RT-PCR), the inactivating effects of soap solutions on rabies viruse in BSR and N2a cells were detected, respectively. Results: This experiment established the BSR or N2a cells model in 24 well cell culture plates, and we found that the upper limit of soap solution concentration which BSR or N2a cells could tolerate was 1%. The inhibitory effect test of different soap solution on rabies virus showed that the 0.5% concentration of soap solution could completely inhibit the survival of CVS-11 strain in both the BSR and N2a cells, but the 0.1% concentration of soap solution could not inhibit the rabies viruses completely. Conclusions The 0.5%-1% concentration of soap solutions could inhibit the survival of CVS-11 strain in three minutes in vivo experiment.

Objective:To explore the risk factors of hypocalcemia and the correlation between calcium supplementation and clinical parameters after parathyroidectomy (PTX) in maintenance hemodialysis patients with secondary hyperparathyroidism (SHPT), and to analyze the effect of calcium supplementation after PTX on the long-term prognosis of patients.Methods:This study was a single-center retrospective study. The patients who underwent PTX in maintenance hemodialysis patients with SHPT in the Huashan Hospital affiliated to Fudan University from October 2014 to March 2021 were retrospectively enrolled. Total PTX with auto transplantation or total PTX alone were the surgical procedures. According to the postoperative requirement of calcium in the first week, the patients were divided into two groups: high calcium supplement (>16.05 g/week) group and low calcium supplement group (≤16.05 g/week). According to the average serum calcium level in the first week after operation, the patients were divided into hypocalcemia group (≤2.1 mmol/L) and non-hypocalcemia group (>2.1 mmol/L) and the differences of clinical parameters between the two groups were compared. The correlation between clinical parameters and the postoperative calcium requirement was examined through Pearson or Spearman correlation analysis. The influencing factors for hypocalcemia after PTX were examined through logistic regression analysis. The survival curve was made by Kaplan-Meier method, and the difference of cumulative survival rate between the two groups was compared by log-rank test.Results:A total of 98 maintenance hemodialysis patients with SHPT were enrolled. The levels of serum calcium, phosphorus, and intact parathyroid hormone (iPTH) after the operation decreased significantly than those of preoperation (all P<0.05). Multiple linear regression analysis showed age ( β=-0.160, P=0.030), iPTH ( β=0.004, P=0.025) and C-reactive protein ( β=0.186, P=0.011) were correlated with postoperative calcium requirement. Preoperative alkaline phosphatase ( OR=1.002, 95% CI 1.000-1.004, P=0.018) and hemoglobin ( OR=0.977, 95% CI 0.954-1.000, P=0.048) independently predicted the occurrence risk of postoperative hypocalcemia through multivariate logistic regression analysis. The recurrence rate of high calcium supplement group was higher than that of low calcium supplement group (10.26% vs 0, P=0.023) and there was no significant difference in all-cause mortality between the two groups (17.95% vs 5.08%, P=0.086). The recurrence rate between the hypocalcemia group and non-hypocalcemia group was no significantly different (8.3% vs 1.8%, P=0.451) and there was no significant difference in all cause mortality between the two groups (12.5% vs 12.7%, P=1.000). Kaplan-Meier survival curve showed that the cumulative survival rate between the two groups was no significantly different (log-rank test χ2=0.147, P=0.702). Conclusions:PTX is a safe and effective therapeutic method to reduce the level of iPTH and improve the metabolism of calcium and phosphorus in SHPT patients. Age, iPTH and C-reactive protein are correlated with the postoperative requirement of calcium in the first week. Preoperative alkaline phosphatase and hemoglobin are independent risk factors for postoperative hypocalcemia. Correcting preoperative electrolyte disorder, improving infection and anemia can reduce the incidence of hypocalcemia after PTX.

Objective To select and identify the cell line for detecting Enterovirus 71 (EV71).Methods pWSK-T7-EV71-GFP containing green fluorescent protein (GFP) gene is an infectious clone for EV71,based on which the UGFP cassette was constructed by inserting GFP gene into 5' and 3' untranslated region (UTR) of the genome of EV71.The lentiviral expression plasmid pLV-UGFP containing UGFP was constructed on the basis of pLV-Puro,a lentiviral vector.To obtain lentivirus,pLV-UGFP plasmids were transfected together with the packaging plasmids into HEK293T cells by liposomes.Then,the target cells BHK-21 were infected with the lentivirus particles.Puromycin-resistant cell colonies were detached from the 6 well-plate and sub-cloned by use of 96 well-plate.Finally,we selected the packaging cell lines that could express the defective replicons stably,named BHK/UGFP cells.Results GFP expression assays indicated that BHK/UGFP cells infected with EV71 could express GFP at 48 hours post-infection,while no green fluorescence was observed after BHK/UGFP cells were infection with Sindbis virus (SINV,XJ-160) or Japanese encephalitis virus (JEV,P3),demonstrating that the selected cells could specifically detect EV71 infection.The sensitive assay results indicated that this method on the basis of BHK/UGFP cells could at least detect 10 PFU/ml of EV71 in tissue culture.Conclusion This result indicated that the BHK/UGFP cells selected in this study were specific and effective to detect EV71 from tissue culture.