The objective of this study was to develop a rapid and precise detection technique for por-cine Senecavirus A(SVA)employing reverse transcription recombinase polymerase amplification(RT-RAA)in conjunction with CRISPR Cas13a technology.Additionally,the study aimed to opti-mize the assay's reaction conditions to enhance amplification efficiency.Eight RT-RAA primer sets were designed based on the conserved gene sequence of porcine SVA,and a series of reaction condi-tions were evaluated to refine the RT-RAA reaction system.Subsequently,CRISPR-derived RNA(crRNA)sequences were developed and selected to construct the RT-RAA-CRISPR reaction sys-tem.The method's specificity was determined by examining six prevalent porcine pathogenic nucleic acids,while its sensitivity was assessed using SVA cRNA standards quantified by digital PCR.The method's stability and the consistency of clinical sample analysis were also evaluated.The findings revealed that the optimized RT-RAA and CRISPR reaction systems exhibited the highest amplifi-cation efficiency at a reaction temperature of 37 ℃.Among the eight crRNAs,five were identified as exhibiting the strongest detection signals.The formulated RT-RAA-CRISPR Cas13a method demonstrated exceptional specificity,showing no cross-reactivity with other common porcine disea-ses,including ASFV,PRRSV,PEDV,PCV2,CSFV,and PRV.The method achieved high sensitivi-ty,detecting as low as 0.86 copies/μL of SVA,and exhibited stable fluorescence output,robust re-producibility,and the ability to complete clinical sample analysis within 50 minutes.Consistency e-valuation with six positive and 58 negative samples indicated 100％agreement in outcomes.These results substantiate that the study successfully established a rapid and specific RT-RAA-CRISPR Cas13a detection method for the on-site identification of porcine Senecavirus A,demonstrating high specificity and sensitivity,and holds promise for application in SVA monitoring and control initia-tives.

The objective of this study was to develop a rapid and precise detection technique for por-cine Senecavirus A(SVA)employing reverse transcription recombinase polymerase amplification(RT-RAA)in conjunction with CRISPR Cas13a technology.Additionally,the study aimed to opti-mize the assay's reaction conditions to enhance amplification efficiency.Eight RT-RAA primer sets were designed based on the conserved gene sequence of porcine SVA,and a series of reaction condi-tions were evaluated to refine the RT-RAA reaction system.Subsequently,CRISPR-derived RNA(crRNA)sequences were developed and selected to construct the RT-RAA-CRISPR reaction sys-tem.The method's specificity was determined by examining six prevalent porcine pathogenic nucleic acids,while its sensitivity was assessed using SVA cRNA standards quantified by digital PCR.The method's stability and the consistency of clinical sample analysis were also evaluated.The findings revealed that the optimized RT-RAA and CRISPR reaction systems exhibited the highest amplifi-cation efficiency at a reaction temperature of 37 ℃.Among the eight crRNAs,five were identified as exhibiting the strongest detection signals.The formulated RT-RAA-CRISPR Cas13a method demonstrated exceptional specificity,showing no cross-reactivity with other common porcine disea-ses,including ASFV,PRRSV,PEDV,PCV2,CSFV,and PRV.The method achieved high sensitivi-ty,detecting as low as 0.86 copies/μL of SVA,and exhibited stable fluorescence output,robust re-producibility,and the ability to complete clinical sample analysis within 50 minutes.Consistency e-valuation with six positive and 58 negative samples indicated 100％agreement in outcomes.These results substantiate that the study successfully established a rapid and specific RT-RAA-CRISPR Cas13a detection method for the on-site identification of porcine Senecavirus A,demonstrating high specificity and sensitivity,and holds promise for application in SVA monitoring and control initia-tives.

Objective To observe the value of ultrasonic biomicroscopy(UBM)imaging for diagnosing chestnut thorn foreign bodies in anterior segment.Methods A total of 70 patients who underwent UBM examination due to chestnut thorn injury or residue of chestnut thorn after removal and then operation were retrospectively enrolled.The distance of chestnut thorn foreign bodies from corneal limbus,and the subconjunctival depth and length of them were measured based on UBM imaging.The efficacy of UBM for diagnosing chestnut thorn foreign bodies in anterior segment was evaluated according to surgical findings,and the correlation of quantitative parameters of foreign bodies were analyzed.Results Among 70 cases of chestnut thorn foreign bodies in anterior segment,single ocular tissue(5/70,7.14％)involvement was detected in 5 cases,while multiple ocular tissues involvement(65/70,92.86％)were noticed in 65 cases according to UBM.Totally 109 foreign bodies were detected by UBM,among which 98 were consistent with surgical findings,with the diagnostic accuracy of 89.91％(98/109),while 11 foreign bodies were not found during operation,with the false positive rate of 10.09％(11/109).Meanwhile,4 foreign bodies found and removed with operation were not displayed with UBM,and the false negative rate of UBM was 3.92％(4/102).Among 98 chestnut thorn foreign bodies correctly diagnosed with UBM,10 located in stromal layer of cornea(10/98,10.20％)and 88 located in ocular wall(88/98,89.80％),including 43 subconjunctival(43/88,48.86％)and 45 sclerotic(45/88,51.14％)foreign bodies,with the median length of 0.200 mm.The distance of 88 foreign bodies in ocular wall from corneal limbus was(2.963±1.504)mm,and the subconjunctival depth was(0.785±0.388)mm.The distance of foreign bodies from corneal limbus were positively correlated with their subconjunctival depth(r,=0.361,P＜0.010),while their lengths were not correlated with their distance from corneal limbus and subconjunctival depth(both P＞0.05).Conclusion UBM imaging was valuable for clinical diagnosis and localization of chestnut thorn foreign bodies in anterior segment.

Objective To observe the value of ultrasonic biomicroscopy(UBM)imaging for diagnosing chestnut thorn foreign bodies in anterior segment.Methods A total of 70 patients who underwent UBM examination due to chestnut thorn injury or residue of chestnut thorn after removal and then operation were retrospectively enrolled.The distance of chestnut thorn foreign bodies from corneal limbus,and the subconjunctival depth and length of them were measured based on UBM imaging.The efficacy of UBM for diagnosing chestnut thorn foreign bodies in anterior segment was evaluated according to surgical findings,and the correlation of quantitative parameters of foreign bodies were analyzed.Results Among 70 cases of chestnut thorn foreign bodies in anterior segment,single ocular tissue(5/70,7.14％)involvement was detected in 5 cases,while multiple ocular tissues involvement(65/70,92.86％)were noticed in 65 cases according to UBM.Totally 109 foreign bodies were detected by UBM,among which 98 were consistent with surgical findings,with the diagnostic accuracy of 89.91％(98/109),while 11 foreign bodies were not found during operation,with the false positive rate of 10.09％(11/109).Meanwhile,4 foreign bodies found and removed with operation were not displayed with UBM,and the false negative rate of UBM was 3.92％(4/102).Among 98 chestnut thorn foreign bodies correctly diagnosed with UBM,10 located in stromal layer of cornea(10/98,10.20％)and 88 located in ocular wall(88/98,89.80％),including 43 subconjunctival(43/88,48.86％)and 45 sclerotic(45/88,51.14％)foreign bodies,with the median length of 0.200 mm.The distance of 88 foreign bodies in ocular wall from corneal limbus was(2.963±1.504)mm,and the subconjunctival depth was(0.785±0.388)mm.The distance of foreign bodies from corneal limbus were positively correlated with their subconjunctival depth(r,=0.361,P＜0.010),while their lengths were not correlated with their distance from corneal limbus and subconjunctival depth(both P＞0.05).Conclusion UBM imaging was valuable for clinical diagnosis and localization of chestnut thorn foreign bodies in anterior segment.

Objective:To compare oxytocin combined with ergometrine with oxytocin alone in terms of primary prophylaxis for postpartum hemorrhage (PPH) at the time of cesarean section (CS).Methods:This was a multicenter double-blind randomized controlled interventional study comparing ergometrine combined with oxytocin and oxytocin alone administered at CS. From December 2018 to November 2019, a total of 298 parturients were enrolled in 16 hospitals nationwide. They were randomly divided into experimental group (ergometrine intra-myometrial injection following oxytocin intravenously; 148 cases) and control group (oxytocin intra-myometrial injection following oxytocin intravenously; 150 cases) according to 1∶1 random allocation. The following indexes were compared between the two groups: (1) main index: blood loss 2 hours (h) after delivery; (2) secondary indicators: postpartum blood loss at 6 h and 24 h, placental retention time, incidence of PPH, the proportion of additional use of uterine contraction drugs, hemostatic drugs or other hemostatic measures at 2 h and 24 h after delivery, the proportion requiring blood transfusion, and the proportion of prolonged hospital stay due to poor uterine involution; (3) safety indicators: nausea, vomiting, dizziness and other adverse reactions, and blood pressure at each time point of administration.Results:(1) The blood loss at 2 h after delivery in the experimental group [(402±18) ml] was less than that in the control group [(505±18) ml], and the difference was statistically significant ( P<0.05). (2) The blood loss at 6 h and 24 h after delivery in the experimental group were less than those in the control group, and the differences were statistically significant (all P<0.05). There were no significant differences between the two groups in the incidence of PPH, the proportion of additional use of uterine contraction drugs, hemostatic drugs or other hemostatic measures at 2 h and 24 h after delivery, the proportion requiring blood transfusion, and the proportion of prolonged hospital stay due to poor uterine involution (all P>0.05). (3) Adverse reactions occurred in 2 cases (1.4%, 2/148) in the experimental group and 1 case (0.7%, 1/150) in the control group. There was no significant difference between the two groups ( P>0.05). The systolic blood pressure within 2.0 h and diastolic blood pressure within 1.5 h of drug administration in the experimental group were higher than those in the control group, and the differences were statistically significant ( P<0.05), but the blood pressure of the two groups were in the normal range. Conclusion:The use of ergometrine injection in CS could reduce the amount of PPH, which is safe and feasible.

Objective: To further observe the efficacy of combined transplantation of islet and bone marrow mesenchymal stem cells (BMSC) in diabetic rats, PET-CT was used to trace cells in vivo to determine the homing and distribution of cells in vivo. Methods: Streptozotocin (STZ)was used to construct a rat model of diabetes mellitus. BMSC could be isolated and cultured by full adherence method; islets were isolated by collagenase; Islets and BMSC were labeled with 18F-FDG in vitro. Diabetic rats were randomly divided into 4 groups, 15 rats in each group: A, Control group; B, Stem cell transplantation group; C, Islet Transplantation group; D, Combined transplantation group, a total of four groups, all transplanted through portal vein, PET-CT tracing the distribution of cells transplanted into the body.7 days after transplantation, the livers of each group were taken, and the homing and distribution of transplanted cells were detected by immunofluorescent staining.The SUV was calculated by the analysis of variance of random block, and the difference between groups was compared by t-test. Results: PET-CT results showed that BMSC were mainly distributed uniformly in the right liver, and the islets of the pancreas were mainly clustered in terminal branches of hepatic portal vein, and BMSC were around the islets of pancreas, but there was no obvious development in the liver of the control group. Conclusions PET-CT can directly reveal the distribution of islets and BMSC in liver after transplantation through portal vein.

Objective:To investigate the effect of short term and intensive insulin pump on the blood lipid and glucose levels of patients with type 2 diabetes.Methods:76 patients with type 2 diabetes were selected and divided into two groups according to different administration.The control group (38 cases) was given routine insulin aspart treatment.The observation group (38 cases) was treated by insulin pump.The blood and lipid indexes and ADL score before treatment,at 1 week and 2 weeks after treatment were compared between two groups.Results:After treatment,the levels of FPG,HbAlc,TC,TG and LDL-C were lower than those before treatment and the level of HDL-C was increased in both groups.The levels ofFPG,HbAlc,TC,TG,LDL-C of observation group were lower than those of the control group,and the level of HDL-C of observation group was higher than that of control group (P＜0.05).At 1 week,2 weeks after treatment,the ADL scores of both groups were significantly improved,and the ADL score of observation group was higher than that of the control group (P＜0.05).Conclusion:Short term intensive insulin pump could significantly improve the blood glucose and lipid metabolism disorders in patients with type 2 diabetes,and improve the ability of daily life.

Objective To investigate the clinical efficacy and safety of sitagliptin combined with metformin in treating patients with type 2 diabetes with non-alcoholic fatty liver diease (NAFLD).Methods Sixty-five patients with type 2 diabetes with non-alcoholic fatty liver admitted into our hospital from June 2012 to June 2015 were randomly divided into two groups.Thirty-three patients in control group were treated with metformin only,and thirty-two patients in observation were treated with sitagliptin combined with metformin.The changes in body mass index (BMI),fasting plasma glucose (FPG),2-hour postprandial glucose (2hPG),Hemoglobin Ale (HbAlc),homeostasis model assessment of insulin resistance (HOMA-IR),and liver function index including that aminopherase of alanine acid (ALT),aminopherase of aspartic acid (AST),gamma-glutamyl transterase (GGT),triglyceride (TG),and total cholesterol (TC) were detected before and after 16 weeks of treatment.And the total effective rate and safety were analyzed and compared.Results After treatment,HOMA-IR,FPG,2hPG,HbA1c,ALT,AST,GGT,TG,and TC were remarkable improvement (P ＜ 0.05),and HOMA-IR,HbA1c,AST,ALT,GGT,and TG were significantly better than those of control (P ＜ 0.05).Additionally,the total effective rate in observation group with 90.9％ was significantly higher than that of control group with 42.4％ (P ＜ 0.05).And difference in incidences of adverse effect between observation group with 15.6％ and control group with 9.1％ were not significant.Conclusion Sitagliptin combined with metformin treatment had a good ability on treating patients with type 2 diabetes with NAFLD by improvement of metabolic syndrome and liver function.

[Abstract ] Objective Globular C1q receptor (gC1qR), a highly acidic receptor protein, is expressed in almost all mamma-lian cells in addition to exoerythrocytic, which can mediate a variety of biological responses.The study aimed to explore gC1qR expres-sion in cervical cancer and itd effects on the biological behaviors of human cervical cancer cells. Methods Retrospective analysis was made on 100 cervical tissue samples of patients in Nanjing Maternal and Child Health Hospital from August 2014 to April 2015, in-cluding 50 cervicitis tissues and 50 cervical cancer tissues.Immunohistochemical SP method was applied in the research of cervical cancer cell line C33a to detect the expression and the location of gC1qR in cervical tissues.Real-time PCR and Western blot analysis were respectively applied to detect the levels of gC1qR mRNA and gC1qR protein expression.Besides, the abilities of C33a cells mi-gration, invasion and apoptosis were respectively assessed by in vitro cell wound healing experiment, transwell assay and flow cytome-try. Results The expression of gC1qR gene was dramatically decreased in the group of cervical cancer tissues when compared with chronic cervictis group (2.18 ±0.37 vs 7.23 ±0.69, 0.27 ±0.09 vs 0.74 ±0.02, P<0.001).gC1qR overexpression could result in sig-nificant up-regulation of cervical celluer apoptosis([22.89 ±1.46]%vs [12.98 ±0.57]%) and down-regulation in migration ([42.60 ± 3.29]%vs [141.83 ±4.71]%) and invasion([26.20 ±2.89]%vs [67.13 ±0.95]%).Typical apoptosis was also observed in cervi-
cal cells by transmission electron microscope. Conclusion gC1qR expression might play an important role in inhibiting the invasion and migration of cervical cancer cells and inducing the apoptosis of cervical carcinoma cells, which provides new clues and potential targets for the treatment of cervical cancer in further research.

Objective To discuss the effectiveness between arthroscope cleaning combined with exercise therapy and exercise therapy in senile osteoarthritis patients.Methods Two hundred and ten senile osteoarthritis patients were divided into operation group and non-operation group by random digits table method with 105 cases each.Patients in operation group received the arthroscope cleaning and exercise therapy,and patients in non-operation group received the exercise therapy alone.The visual analogue score (VAS) and international knee documentation committee knee evaluation form (IKDC) score before treatment and 1,6 and 12 months after treatment were compared.Results A total of 210 patients received the analysis of result without drop-out.There were no statistical differences in VAS and IKDC scores before treatment and 6 and 12 months after treatment between operation group and non-operation group (P ＞ 0.05).The VAS and IKDC scores 1,6,12 months after treatment were significantly improved compared with those before treatment in operation group,and there were statistical differences (P ＜ 0.05).The VAS and IKDC scores 6,12 months after treatment were significantly improved compared with those before treatment in non-operation group,and there were statistical differences (P ＜ 0.05).According to the IKDC score,in operation group 15 patients were excellent,61 patients were fine,19 patients were general and 10 patients were bad after treatment,and good effectiveness rate was 72.38％ (76/105).In non-operation group,14 patients were excellent,59 patients were fine,20 patients were general and 12 patients were bad after treatment,and good effectiveness rate was 69.52％ (73/105).There was no statistical difference in the good effectiveness rate between 2 groups (P ＞ 0.05).Conclusions Exercise therapy with guidance can significantly improve the function of senile knee joint osteoarthritis,and ease joint pain symptoms.Arthroscope cleaning combined with exercise therapy can relieve the joint acute inflammatory symptoms,but it has no significant clinical effectiveness in long-term treatment of osteoarthritis.