Objective To overcome the limitations of existing human respiratory syncytial virus(hRSV)animal models,such as semi-permissiveness and short duration of infection,this study established an IL2rg gene knockout(IL2rg-/-)rat model using TALEN gene editing technology.Methods The animal model was infected with hRSV intranasally.Clinical characteristics,body weight,and temperature changes were observed over the infection period(0～35 days).The total viral loads in respiratory organs,such as the nasal tissue,trachea,and lungs,were measured at various time points(4,11,20,and 35 days post-infection).Pathological analysis was conducted on target organs at the endpoint of observation(35 days post-infection).Changes in peripheral blood T,B,NK,and NKT cells and various cytokines were assessed at various time points(4,20,and 35 days post-infection).Results(1)IL2rg/-knockout rats sustained high viral loads in the nasal cavity upon intranasal inoculation with hRSV.The average peak titer rapidly reached 1 × 1010 copies/g in nasal tissue and 1 × 107 copies/g up to 5 weeks post-infection.(2)However,no significant pathological changes were noted in nasal,tracheal,or lung tissues.(3)An increase was observed in the content of peripheral blood B cells in hRSV-infected IL2rg--rats.(4)IL-6 and MCP-1 were increased in the early stage of infection and then decreased at the end of the observation period.Conclusions This study established a new IL2rg-/-rat model using TALEN technology and found that this model effectively supported high-level replication and long-term infection of hRSV,providing a good basis for antiviral drug screening and in vivo efficacy evaluation of anti-hRSV antibodies.

Objective:To investigate influences of curcumin on LPS-induced autophagy in corneal stromal cells by regulating phosphatidylinositol 3-kinase(PI3K)/serine-threonine kinase(AKT)/mammalian target of rapamycin(mTOR)signaling pathway.Methods:Mouse corneal stromal cells were grouped into control group,LPS group(1 μg/μl LPS),L-Cur group(15 μmol/L curcumin),M-Cur group(30 μmol/L curcumin),H-Cur group(50 μmol/L curcumin),and H-Cur+740Y-P group(50 μmol/L curcumin+10 μmol/L PI3K activator 740Y-P).MTT was applied to detect effect of curcumin on corneal stromal cytotoxicity and cell viability;ELISA was applied to detect levels of inflammatory factors in corneal stromal cells;immunofluorescence staining was applied to detect levels of Beclin1 and LC3 in mouse corneal stromal cells;Western blot was applied to detect expressions of autophagy-related proteins and pathway-related proteins in corneal stromal cells.Results:0～90 μmol/L curcumin had no obvious toxicity to corneal stromal cells.Compared with control group,A570 nm,levels of Beclin1 and LC3Ⅱ/Ⅰ in LPS group were obviously decreased(P<0.05),levels of IL-6,IL-8,p-PI3K/PI3K,p-AKT/AKT and p-mTOR/mTOR were obviously increased(P<0.05);compared with LPS group,A570 nm,levels of Beclin1 and LC3Ⅱ/Ⅰ in L-Cur group,M-Cur group,H-Cur group were obviously increased in turn(P<0.05),levels of IL-6,IL-8,p-PI3K/PI3K,p-AKT/AKT and p-mTOR/mTOR were obviously decreased in turn(P<0.05);H-Cur+740Y-P eliminated beneficial effect of curcumin on corneal stromal cells.Conclusion:Curcumin may promote LPS-induced autophagy in corneal stromal cells by down-regulating PI3K/AKT/mTOR signaling pathway.

Objective:To investigate the effect and mechanism of miR-195 regulating FOXK1 gene and PI3K/Akt pathway on stomach adenocarcinoma proliferation, invasion and migration ability.Methods:Public database samples were employed to analyze the expression differences and prognostic significance of miR-195 in stomach adenocarcinoma. After overexpression of mir-195-5p in two cell lines, MGC803 and AGS, altered cell proliferation, invasion, and migration abilities were detected by Alamar Blue, Wound healing, and Transwell assays. The potential target genes and binding sites of miR-195 were predicted by the starBase. Western blot was used to detect the expression levels of foxk1 and phosphorylation sites in the PI3K/Akt pathway of target genes after overexpression of mir-195-5p. A Dual-luciferase reporter assay was used to verify the relationship between mir-195-5p and foxk1. Statistical analyses were performed with IBM SPSS 22 software and R 4.0.3.Results:Our results showed a significant over-expression of miR-195 in the tumor tissues, compared with the paired normal tissues ( P<0.001) , which could inhibit the proliferation and invasion of stomach carcinoma cells and significantly correlated with survival ( P=0.011) . Moreover, our study indicated that miR-195 depressed the expression of FOXK1 and significantly reduced the activation of the PI3K/Akt pathway, which had a negative effect on the proliferation and invasion of stomach carcinoma cells. The phosphorylated Akt (s473 site) expression in the PI3K/Akt pathway was significantly decreased after overexpression of miR-195. Conclusion:Overall, our studies clarify the important function of the miR-195 in the diagnosis and therapy of patients with stomach carcinoma and reveal the FOXK1 and PI3K/Akt pathway regulation by the miR-195, which are of important clinical significance in the differential diagnosis.

Objective:To evaluate the value of transrectal ultrasound(TRUS)in diagnosing extramural venous invasion(EMVI) of rectal cancer.Methods:Clinical data of 81 rectal cancer patients were retrospectively analyzed in the People′s Hospital of Inner Mongolia Autonomous Region from January 2015 to December 2021. The extramural vascular sonographic features in these patients were summarized. Based on the postoperative pathology and compared with MRI examination, the efficacy of TRUS for the diagnosis of EMVI in rectal cancer was investigated.Results:①According to the sonographic presentation, extramural vessels of rectal cancer can be divided into the following four types: typeⅠshowed that the mass broke through the muscularis propria of the intestinal wall, and no peripheral vascular shadow was observed; typeⅡshowed that the mass broke through the muscularis propria, surrounded by extraneous vessels with normal diameter, running and blood flow; type Ⅲ showed that the mass broke through the intrinsic muscle layer of the intestinal wall and was surrounded by extramural vessels with abnormally thickened diameters, and blood flow filling defects; type Ⅳ showed a mass that broke through the intrinsic muscular layer, with irregularly dilated vessels outside the peritumoral wall and no blood flow signal in the lumen. ②With type Ⅲ and type Ⅳ as positive signs of EMVI, the diagnosis compliance rate of TRUS was 90.1% (Kappa=0.580, P<0.001), sensitivity was 58.3%, specificity was 95.7%, positive predictive value was 70.0%, negative predictive value was 93.0%, and area under the ROC curve(AUC) was 0.770. The diagnosis compliance rate of MRI was 86.4% (Kappa=0.541, P<0.001), sensitivity was 75.0%, specificity was 88.4%, positive predictive value was 52.9%, negative predictive value was 95.3%, and the AUC was 0.817. The differences in sensitivity and specificity between TRUS and MRI for the diagnosis of rectal cancer EMVI were not statistically significant ( P>0.05), and the differences in the AUC were not statistically significant ( Z=0.447, P=0.655). Conclusions:TRUS is valuable in assessing the extramural vascular status of rectal cancer and is expected to be an effective imaging method for preoperative diagnosis of EMVI.

Objective:To discuss the surgical effect of modified cervical cerclage for the treatment of pregnant women with cervical insufficiency.Methods:The clinical data of 225 pregnant women who underwent modified cervical cerclage in Qilu Hospital (Qingdao) were selected for retrospective analysis from April 2014 to June 2020. Surgical success rate, full-term birth rate, preterm birth rate, prolonged pregnancy weeks and newborn birth weight were compared between singleton and twin pregnancies, preventive cerclage and emergency cerclage, surgery before and after 18 weeks, naturally and in vitro fertilization and embryo transfer (IVF-ET) conceived pregnant women respectively.Results:Among the 225 pregnant women, the gestational weeks of surgery were 14-24 +5 weeks, mean gestational weeks of delivery were 38 +2 weeks (35 +5-39 +3 weeks), the number of prolonged gestation were (20.3±5.2) weeks, and the newborn birth weight was (3 065±735) g; the overall surgical success rate was 92.9% (209/225), and the miscarriage rate was 7.1% (16/225); among the surviving newborns, the full-term birth rate was 73.7% (154/209), and the preterm birth rate was 26.3% (55/209). All cases had no intraoperative complications. Among the 225 pregnant women, 202 (89.8%, 202/225) cases were singleton pregnancies, and 23 (10.2%, 23/225) cases were twin pregnancies; 201 (89.3%, 201/225) cases underwent preventive cervical cerclage, and 24 (10.7%, 24/225) cases underwent emergency cervical cerclage; 190 (84.4%, 190/225) cases underwent the surgery before 18 weeks, and 35 (15.6%, 35/225) cases underwent the surgery after 18 weeks; 49 (21.8%, 49/225) cases were conceived by IVF-ET. There was no statistically significant difference in the overall surgical success rate of single and twin group ( P>0.05). The full-term birth rate, newborn birth weight and prolonged pregnancy weeks of single group were higher than those of twin group ( P<0.05). There were no statistical differences between preventive and emergency cerclage in overall surgical success rate, full-term birth rate, preterm birth rate, and newborn birth weight (all P>0.05). The pregnancy prolonged weeks of preventive cerclage was higher than that of emergency cerclage ( P<0.05). There were no statistically significant differences in the overall surgical success rate, full-term birth rate, preterm birth rate and birth weight of newborns at different surgical timings (all P>0.05). The pregnancy prolonged week for those who underwent surgery before 18 weeks was higher than that of surgery after 18 weeks ( P<0.05). The premature birth rate of IVF-ET was higher than that of naturally conceived pregnant women ( P<0.05). Conclusion:The modified cervical cerclage could effectively prolong the gestational weeks of delivery, reduce the rate of preterm birth, and the operation is simple and easy to promote. It could be used as a surgical option for patients with cervical insufficiency.

Objective:To analyze, explore and evaluate the clinical characteristics, abnormal thyroid function and follow-up of anti-hyperthyroidism treatment mode in patients with hyperthyroidism (commonly abbreviated as HT) combined with liver injury.Methods:The clinical data of patients with hyperthyroidism combined with liver injury were retrospectively analyzed, and then patients were divided into treated and untreated group according to whether they received anti-hyperthyroidism treatment before the consultation. Patients’ thyroid and liver function test indicators at the time of treatment were analyzed to determine the main cause of liver injury. The characteristics of liver injury were analyzed in the treatment group. Patients with severe thyroid toxicity and hyperthyroidism combined with liver injury were followed-up with anti-hyperthyroid therapy, mainly low-dose methimazole (MMI) and radioactive iodine therapy to evaluate its efficacy and safety. The comparison between data groups was performed by t-test, rank sum test and χ2 test. Results:Among the 43 cases with hyperthyroidism combined with liver injury, 19 were males and 24 were females, aged 49.0 ± 14.6 years-old; 16 cases (16/43, 37.21%) aged 40 to≤60 years- old, and 15 cases (15/43, 34.88%) aged > 60 years-old. There were 22 untreated cases (untreated group, accounting for 51.16%), and 21 treated cases with anti-hyperthyroidism (treatment group, accounting for 48.84%) at the time of consultation. Thyroid function indicators (FT3, FT4, TSH) and liver function indicators (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, γ-glutamyltransferase, total bilirubin) of the two groups were compared, and the difference was not statistically significant ( P > 0.05). The order of liver injury from mild to severe in patients with different treatment options were: methimazole (MMI) < propylthiouracil < radioactive iodine

Objective To evaluate the effect of Xingqi Tongfu Pad on the intestinal functions of children with acute abdomen after surgery. Methods The random digit table was used to randomize 146 patients with acute abdomen into treatment group (n=71) and control group (n=75) according to their registration number. In the control group, the patients only received normal treatment and nursing, while those in the treatment group received Xingqi Tongfu Pad for the acupoint application in addition to routine treatments. The two groups were compared in terms of the time of bowel sound recovery , the exhaust defecation time and gastrointestinal dysfunction score. Results The bowel sounds and the first exhaust and defecation in the treatment group appeared obviously earlier than those in the control group . The score on gastrointestinal dysfunction in the treated group was significantly higher than that of the control group (P<0.05). Conclusions Xingqi Tongfu Pad can promote the recovery of gastrointestinal function after abdominal operation. It deserves popularization for its positive effect, cheap price and easy operation.

Objective:To study the changes and significance of Th17 and Treg cells as well as their associated cytokines in pe-ripheral blood of patients with chronic hepatitis C and cirrhosis in order to investigate their role in the pathogenesis of chronic hepatitis C and hepatitis C cirrhosis. Methods:Flow cytometry and ELSIA assay were used to detect the expression ratio of Th17 and Treg cells and the serum levels of IL-10,TGF-β,IL-6 and IL-17 in peripheral blood of healthy human and patients with chronic hepatitis C and hepatitis C cirrhosis. Than we analyzed the differences of the above detection index between the healthy and patients with chronic hepatitis C and hepatitis C cirrhosis. Results:The expression ratio of Th17 cells (1. 33%±0. 30%) in peripheral blood of patients with chronic hepatitis C and IL-6[(8. 10±2. 42)ng/L] and IL-17[(16. 70±4. 73)ng/L] serum levels were significantly higher than those in the healthy (Th17:1. 14%±0. 19%) and IL-6[(1. 72±6. 70)ng/L],IL-17[(12. 29±1. 88)ng/L],P<0. 05;The expression ratio of Treg cells in peripheral blood of patients with hepatitis C cirrhosis and chronic hepatitis C patients was significantly higher than that in the healthy (6. 21%±0. 76%,5. 89%±0. 85% vs 5. 51%±0. 59% ),P<0. 05,The ratio of Th17/Treg in patients with cirrhosis was significantly lower than that in patients with chronic hepatitis C and the healthy(0. 19±0. 02 vs 0. 22±0. 03,0. 21±0. 03),P<0. 05;the levels of IL-1[(16. 21±3. 76)ng/L] and TGF-β[(5. 15±0. 83)ng/L] in peripheral blood of patients with hepatitis C cirrhosis were significantly higher than those in chronic hepatitis C[(14. 36±2. 78)ng/L;(4. 47±0. 87)ng/L] and the healthy[(14. 01±3. 01)ng/L;(4. 43±0. 98)ng/L)],P<0. 05. Conclusion:Th17 and Treg cells and their related cytokines involved in the process of chronicity of hepatitis C and cirrhosis,Th17 and Treg cells and their related cytokines in the treatment and prevention of chronicity of hepatitis C and cirrhosis may have important significance.

Objective To encode the protein cystatin M with CST6 gene and construct a CST6-overexpression vector,and transfect it into the gastric cancer cells SGC-7901 in order to observe the effect of cystatin M on the proliferation and migration abilities of SGC-7901 cells.Methods There were three groups in the experiment:pcDNA3.1(+)-CST6 group,pcDNA3.1 (+)group and non-transfected group.RT-PCR and Western blot were used to identify the RNA expression of CST6 in SGC-7901/CST6 cells.The proliferation and migration abilities of the transfected cells were detected by MTT and cell wound healing assay,respectively.Results SGC-7901/CST6 cells stably expressed CST6 gene RNA and cystatin M protein.The proliferation and migration of pcDNA3 .1 (+)-CST6 group cells were reduced compared with those of cells in the other two groups (P<0.05).Conclusion Cystatin M can inhibit the proliferation and migration of SGC-7901 cells.