Objective:To establish a Zika virus-infected suckling SD rat model with type Ⅰ interferon receptor deficiency（SD-Ifnar1 -/-［cc］）and provide a novel animal model for investigating Zika virus pathogenesis and developing therapeutic strategies. Methods:Seventeen-day-old male SD-Ifnar1 -/-［cc］rat pups were randomly divided into experimental and control groups（ n=6）. The experimental group received an intraperitoneal injection of Zika virus strain SZ-wiv01（5×10 4 PFU/rat，200 μl），while the control group received an equal volume of phosphate-buffered saline（PBS）. Animals were euthanized via CO 2 asphyxiation on days 12 and 15 post-infection（dpi），and brain，spleen，liver，and testis tissues were harvested. Viral loads and cytokine expression levels were quantified using quantitative real-time PCR qRT-PCR），and histopathological evaluation was performed via HE staining. Results:qRT-PCR analysis revealed no detectable Zika virus RNA（Ct >35）in control tissues. In the experimental group，viral RNA（Ct <35）was detected in the brain，spleen，liver，and testis by day 12，with stable viral loads across tissues by day 15（ P>0.05）. Cytokine profiling demonstrated significant upregulation in the brain at day 12：IFN-β（5.58-fold， P<0.01），IL-6（7.11-fold， P<0.01），and CCL5（3.79-fold， P<0.01）. By day 15，these levels remained elevated（IFN-β：3.07-fold；IL-6：4.04-fold；CCL5：5.22-fold；all P<0.01）. In the liver，IFN-β mRNA decreased to 20% of the control level by day 15（ P<0.05），while IL-6 declined to 24% and CCL5 mRNA dropped to 38% by day 12. No significant cytokine changes were observed in the spleen（ P>0.05）. Testicular tissues exhibited reduced IFN-β mRNA levels（43% of control at day 12，31% at day 15； P<0.05）. Histopathological analysis revealed marked splenomegaly with disrupted splenic corpuscle architecture and lymphocyte depletion，significant inflammatory cell infiltration in hepatic portal areas，and testicular structural disorganization with inflammatory infiltration in Zika-infected rats. Conclusions:The SD-Ifnar1 -/-［cc］suckling rat model is successfully established and validated. This model recapitulates systemic Zika virus infection，tissue-specific pathology，and immune response dynamics，providing a robust platform for elucidating viral pathogenesis and advancing antiviral drug development.

AIM:To establish a cell model of visceral hypersensitivity and nerve hyperplasia in irritable bowel syndrome(IBS)by conducting an in vitro co-culture of mouse P815 mast cells and N2a nerve cells and explore its possible mechanism.METHODS:Enzyme-linked immunosorbent assay(ELISA)with three replicates was used to confirm the C48/80-induced P815 degranulation.The length of neurites was observed under bright field microscopy to determine the number of differentiated neurons,thereby selecting the concentration of retinoic acid(RA)for stimulating the differentia-tion of N2a cells,with four replicates.A co-culture system of P815 and N2a cells was established using Transwell cham-bers with four replicates.The following groups were established:N2a cells cultured alone,N2a cells co-cultured with P815 cells,N2a cells co-cultured with P815 cells plus C48/80,and N2a cells plus RA group.After co-culturing,the num-ber of differentiated N2a cells was observed under bright field.The expression of nerve growth factor(NGF),tyrosine ki-nase receptor A(TRKA),growth-associated protein-43(GAP43),neuron-specific enolase(NSE),synapsin(SYN),and postsynaptic density protein-95(PSD-95)at both protein and gene levels in N2a cells was detected using Western blot and polymerase chain reaction(PCR),with four replicates.RESULTS:The best condition for N2a differentiation was stimulation with 10 μmol/L RA for 24 hours,whereas the best condition for degranulation was stimulation of P815 cells with 20 mg/L C48/80 for 24 hours.Compared with N2a cells cultured alone,the differentiation ratio of the N2a+P815+C48/80 and N2a+RA groups was significantly increased(P<0.01),and the protein and mRNA expressions of NGF,TRKA,GAP43,NSE,SYN,and PSD-95 were significantly increased(P<0.05).CONCLUSION:Our results revealed that mast cell degranulation enhances the level of nerve hyperplasia in enteric nerve cells and promotes changes in nerve structure and function.Synaptic remodeling regulated by abnormal expression of key proteins such as NGF,TRKA,and GAP43 is involved in the nerve hyperplasia induced by mast cell degranulation.

AIM:To establish a cell model of visceral hypersensitivity and nerve hyperplasia in irritable bowel syndrome(IBS)by conducting an in vitro co-culture of mouse P815 mast cells and N2a nerve cells and explore its possible mechanism.METHODS:Enzyme-linked immunosorbent assay(ELISA)with three replicates was used to confirm the C48/80-induced P815 degranulation.The length of neurites was observed under bright field microscopy to determine the number of differentiated neurons,thereby selecting the concentration of retinoic acid(RA)for stimulating the differentia-tion of N2a cells,with four replicates.A co-culture system of P815 and N2a cells was established using Transwell cham-bers with four replicates.The following groups were established:N2a cells cultured alone,N2a cells co-cultured with P815 cells,N2a cells co-cultured with P815 cells plus C48/80,and N2a cells plus RA group.After co-culturing,the num-ber of differentiated N2a cells was observed under bright field.The expression of nerve growth factor(NGF),tyrosine ki-nase receptor A(TRKA),growth-associated protein-43(GAP43),neuron-specific enolase(NSE),synapsin(SYN),and postsynaptic density protein-95(PSD-95)at both protein and gene levels in N2a cells was detected using Western blot and polymerase chain reaction(PCR),with four replicates.RESULTS:The best condition for N2a differentiation was stimulation with 10 μmol/L RA for 24 hours,whereas the best condition for degranulation was stimulation of P815 cells with 20 mg/L C48/80 for 24 hours.Compared with N2a cells cultured alone,the differentiation ratio of the N2a+P815+C48/80 and N2a+RA groups was significantly increased(P<0.01),and the protein and mRNA expressions of NGF,TRKA,GAP43,NSE,SYN,and PSD-95 were significantly increased(P<0.05).CONCLUSION:Our results revealed that mast cell degranulation enhances the level of nerve hyperplasia in enteric nerve cells and promotes changes in nerve structure and function.Synaptic remodeling regulated by abnormal expression of key proteins such as NGF,TRKA,and GAP43 is involved in the nerve hyperplasia induced by mast cell degranulation.

Objective:To establish a Zika virus-infected suckling SD rat model with type Ⅰ interferon receptor deficiency（SD-Ifnar1 -/-［cc］）and provide a novel animal model for investigating Zika virus pathogenesis and developing therapeutic strategies. Methods:Seventeen-day-old male SD-Ifnar1 -/-［cc］rat pups were randomly divided into experimental and control groups（ n=6）. The experimental group received an intraperitoneal injection of Zika virus strain SZ-wiv01（5×10 4 PFU/rat，200 μl），while the control group received an equal volume of phosphate-buffered saline（PBS）. Animals were euthanized via CO 2 asphyxiation on days 12 and 15 post-infection（dpi），and brain，spleen，liver，and testis tissues were harvested. Viral loads and cytokine expression levels were quantified using quantitative real-time PCR qRT-PCR），and histopathological evaluation was performed via HE staining. Results:qRT-PCR analysis revealed no detectable Zika virus RNA（Ct >35）in control tissues. In the experimental group，viral RNA（Ct <35）was detected in the brain，spleen，liver，and testis by day 12，with stable viral loads across tissues by day 15（ P>0.05）. Cytokine profiling demonstrated significant upregulation in the brain at day 12：IFN-β（5.58-fold， P<0.01），IL-6（7.11-fold， P<0.01），and CCL5（3.79-fold， P<0.01）. By day 15，these levels remained elevated（IFN-β：3.07-fold；IL-6：4.04-fold；CCL5：5.22-fold；all P<0.01）. In the liver，IFN-β mRNA decreased to 20% of the control level by day 15（ P<0.05），while IL-6 declined to 24% and CCL5 mRNA dropped to 38% by day 12. No significant cytokine changes were observed in the spleen（ P>0.05）. Testicular tissues exhibited reduced IFN-β mRNA levels（43% of control at day 12，31% at day 15； P<0.05）. Histopathological analysis revealed marked splenomegaly with disrupted splenic corpuscle architecture and lymphocyte depletion，significant inflammatory cell infiltration in hepatic portal areas，and testicular structural disorganization with inflammatory infiltration in Zika-infected rats. Conclusions:The SD-Ifnar1 -/-［cc］suckling rat model is successfully established and validated. This model recapitulates systemic Zika virus infection，tissue-specific pathology，and immune response dynamics，providing a robust platform for elucidating viral pathogenesis and advancing antiviral drug development.

Objective:To explore the effects of different courses of hyperbaric oxygen (HBO) therapy on the clinical efficacy, neurological function, and negative emotions of patients with spinal cord injury.Methods:The clinical data of 126 patients with spinal cord injury treated in the Department of Orthopedics of Changzheng Hospital Affiliated to Naval Medical University and the Department of Orthopedics of Shanghai Kaiyuan Orthopedic Hospital from January 2017 to October 2019 were retrospectively analyzed. According to whether or not the patients received HBO therapy after surgery and different courses of HBO therapy, the patients who did not receive HBO therapy were taken into the control group ( n=32), the patients who received 1-2 courses of HBO therapy were taken into the observation group A ( n=32), the patients who received 3-4 courses of treatment were taken into the observation group B ( n=31), and the patients who received 5-6 courses of treatment were taken into the observation group C ( n=31). All patients were evaluated by the American Spinal Injury Association (ASIA) Impairment Scale, functional independence measure (FIM), zung self-rating anxiety scale (SAS), and zung self-rating depression scale (SDS) before and after treatment to assess the clinical efficacy. Results:After treatment, the scores of ASIA pin prick and light touch sensation in four groups were significantly improved compared with those before treatment ( P<0.05), the improvements of observation group A, B, and C were better than those of the control group, and the improvements of observation group B and C were better than those of the observation group A ( P<0.05). After treatment, ASIA motor scores and FIM scores of the four groups were significantly improved compared with those before treatment, and the differences were statistically significant ( P<0.05). After treatment, ASIA motor scores and FIM scores of observation group A, B, and C were higher than those of the control group, and the improvements of observation group B and C were better than those of the observation group A, with statistically significant differences ( P<0.05). After treatment, SAS scores and SDS scores of the four groups were significantly decreased compared with those before treatment, and the differences were statistically significant ( P < 0.05). The scores of SAS and SDS in the observation group A, B, and C after treatment were lower than those of the control group, and the differences were statistically significant ( P<0.05). Conclusion:Hyperbaric oxygen can effectively alleviate the spinal cord injury symptoms of patients, improve their neurological function, and reduce their negative emotions, and can exert better therapeutic effect after 3-4 treatment courses or more.

Objective:To explore the effects of different courses of hyperbaric oxygen (HBO) therapy on the clinical efficacy, neurological function, and negative emotions of patients with spinal cord injury.Methods:The clinical data of 126 patients with spinal cord injury treated in the Department of Orthopedics of Changzheng Hospital Affiliated to Naval Medical University and the Department of Orthopedics of Shanghai Kaiyuan Orthopedic Hospital from January 2017 to October 2019 were retrospectively analyzed. According to whether or not the patients received HBO therapy after surgery and different courses of HBO therapy, the patients who did not receive HBO therapy were taken into the control group ( n=32), the patients who received 1-2 courses of HBO therapy were taken into the observation group A ( n=32), the patients who received 3-4 courses of treatment were taken into the observation group B ( n=31), and the patients who received 5-6 courses of treatment were taken into the observation group C ( n=31). All patients were evaluated by the American Spinal Injury Association (ASIA) Impairment Scale, functional independence measure (FIM), zung self-rating anxiety scale (SAS), and zung self-rating depression scale (SDS) before and after treatment to assess the clinical efficacy. Results:After treatment, the scores of ASIA pin prick and light touch sensation in four groups were significantly improved compared with those before treatment ( P<0.05), the improvements of observation group A, B, and C were better than those of the control group, and the improvements of observation group B and C were better than those of the observation group A ( P<0.05). After treatment, ASIA motor scores and FIM scores of the four groups were significantly improved compared with those before treatment, and the differences were statistically significant ( P<0.05). After treatment, ASIA motor scores and FIM scores of observation group A, B, and C were higher than those of the control group, and the improvements of observation group B and C were better than those of the observation group A, with statistically significant differences ( P<0.05). After treatment, SAS scores and SDS scores of the four groups were significantly decreased compared with those before treatment, and the differences were statistically significant ( P < 0.05). The scores of SAS and SDS in the observation group A, B, and C after treatment were lower than those of the control group, and the differences were statistically significant ( P<0.05). Conclusion:Hyperbaric oxygen can effectively alleviate the spinal cord injury symptoms of patients, improve their neurological function, and reduce their negative emotions, and can exert better therapeutic effect after 3-4 treatment courses or more.

Objective: To investigate the effect of hypertensive disorder complicating pregnancy (HDCP) on the mortality and early complications of premature infants. Methods: The general clinical data of preterm infants with gestational age 24-36^{+ 6} weeks were collected from the cooperative units in the task group from January 1, 2013 to December 31, 2014.According to the severity of HDCP, the infants were divided into 4 groups: HDCP group, preeclampsia group, eclampsia group and non HDCP group, the mortality and major complications of preterm infants were compared, and the influencing factors were analyzed. Results: The mortality rate of preterm in the HDCP group was significantly higher than that of non HDCP group, and there was statistical significance (χ²=9.970, P=0.019). Eclampsia had a highest fatality rate (4.8%) in the early stage, compared with non HDCP group (2.2%), and the difference was statistically significant.Comparison of HDCP group (1.8%) and eclampsia group (3.2%) suggested that there was no statistically significant difference.The incidence of respiratory distress syndrome (RDS) in preterm in HDCP group was significantly higher than that of non HDCP group, and there was statistical significance (χ²=13.241, P=0.004). Eclampsia group showed the highest incidence (35.4%), compared with non HDCP group (16.2%), the difference was statistically significant, but compared with HDCP group (19.9%), preeclampsia group (17.1%), there was no significant diffe-rence.The incidence of bronchopulmonary dysplasia (BPD) in preterm in HDCP group was significantly higher than that of non HDCP group (χ²=9.592, P=0.022), the highest incidence showed up in eclampsia group (9.7%), compared with non HDCP group (2.0%) and HDCP group (1.7%), the difference was statistically significant.But there was no statistically significant difference, compared with preeclampsia group.As the degree of HDCP aggravated, the incidence of BPD gradually rose.There was no significant impact on necrotizing enterocolitis (NEC), retinopathy of prematurity (ROP), intraventricular hemorrhage (IVH) and sepsis of HDCP (χ²=7.054, 7.214, 0.358, 3.852; P=0.070, 0.065, 0.949, 0.278). Considering the overall outcome of the child, that was, whether the child died or survived, he had at least one complication, and HDCP had an effect on it (χ²=15.697, P=0.001), so the incidence increased while the degree of HDCP rose gradually.After adjusting gestational age, birth weight, sex, way of delivery, placental abruption and front placenta, prenatal hormonal, gestational diabetes, neonatal asphyxia and other factors, the results displayed that HDCP was the factor leading to the death of premature baby (OR=2.159, 95%CI: 1.093-4.266), and comparison between preeclampsia and eclampsia showed no statistical difference (P=0.714, 0.389); HDCP had no significant influence on RDS, BDP, ICH, NEC, ROP and sepsis. Conclusions HDCP leads to increased risk of premature death, but also leads to the increased incidence of RDS and BPD, but it had no obvious effect on NEC, ROP, IVH, sepsis and other complications.

Objective To compare the therapeutic effect of combined fascia sheath suspension and frontal muscle flap suspension in the treatment of congenital severe congenital ptosis.Methods 46 cases (58 eyes) with congenital severe upper eyelid prolapse who were treated from March 2015 to March 2016 in Eye Hospital of Hebei Province were selected.The patients were divided into two groups:combined fascial sheath suspension group and frontalis muscle flap suspension.Comparisons of postoperative patients with eyelid height,eyelid radian,corneal exposure with eyes closed,eyelid activity,complication of the situation were carried out.And patient satisfaction survey was conducted.Results The effective rate of combined fascial sheath suspension group was 93.10％,the complication rate was 6.9％,and the patient satisfaction rate was 91.22±0.37,significantly better than the frontal muscle flap suspension group of patients (75.86 ％,24.14 ％ and 75.05 ± 0.28,respectively).The time of closure of the eyelid was shortened,the complications were less,and the eyelid had better activity.The difference between the two groups was statistically significant (P＜0.05).Conclusions Combined fascia sheath for congenital severe upper eyelid droop is significant,with fewer complications and a higher rate of satisfaction,which is worth clinical promotion.

MicroRNAs (miRNAs) play critical roles in the development and progression in various cancers.Dysfunctional miR-9 expression remains ambiguous,and no consensus on the metastatic progression of ovarian cancer has been reached.In this study,results from the bioinformatics analysis show that the 3'-UTR of the E-cadherin mRNA was directly regulated by miR-9.Luciferase reporter assay results confirmed that miR-9 could directly target this 3'-UTR.miR-9 and E-cadherin expression in ovarian cancer tissue was quantified by qRTPCR.Migration and invasion were detected by wound healing and Transwell system assay in SKOV3 and A2780.qRT-PCR and Western blot were performed to detect the epithelial-mesenchymal transition-associated mRNA and proteins.Immunofluorescence technique was used to analyze the expression and subcellular localization of Ecadherin,N-cadherin,and vimentin.The results showed that miR-9 was frequently upregulated in metastatic serous ovarian cancer tissue compared with paired primary ones.Upregulation of miR-9 could downregulate the expression of E-cadherin but upregulate the expression of mesenchymal markers (N-cadherin and vimentin).Overexpression of miR-9 could promote the cell migration and invasion in ovarian cancer,and these processes could be effectively inhibited via miR-9 inhibitor.Thus,our study demonstrates that miR-9 may promote ovarian cancer metastasis via targeting E-cadherin and a novel potential therapeutic approach to control metastasis of ovarian cancer.

Soladulcidine is a steroidal alkaloid abundant in Solanum dulcamara L. with antitumor and other biological activities. In this study, ten soladulcidine derivatives were synthesized through esterification at C-3-hydroxy group, modification at NH group of F ring of esterification of E ring-opening products. The in vitro antiproliferative activity of these synthesized derivatives against prostate cancer (PC-3) cell line was assessed. Within this series of compounds, compound 19 exhibited the most potent inhibitory effect against the proliferation of PC-3 cell line (IC50=4.80±.9μmol/L).