Objective To explore the biological behavior and mechanism of isovitexin(Isov)on pancreatic cancer cells.Methods Isov was used to treat the human normal pancreatic ductal epithelial cells HPDE and PC cell lines,and CCK-8 was used to detect the cell proliferation and calculate the half inhibitory concentration(IC50).The PC cell line PANC-1 cells were grouped into the control group,the Isov group,the Isov+in-miR-NC group,the Isov+in-miR-339-5p group,the Isov+in-miR-339-5p+si-NC group and the Isov+in-miR-339-5p+si-HSPA8 group.The survival,migration and invasion of PANC-1 cells were detected by CCK-8,scratch healing assay and Transwell assay.Real time fluorescence quantitative PCR was used to detect the mRNA expression of miR-339-5p and heat shock protein family A member 8(HSPA8)in PANC-1 cells.Western blot assay was used to detect protein HSPA8 expression in various groups of cells.Dual luciferase reporter gene was used to detect the targeting effect of miR-339-5p and HSPA8.A xenograft nude mouse model was used to determine the in vivo anticancer effects of Isov.Results Isov inhibited PC cell proliferation but had little cytotoxicity to HPDE cells.Isov could obviously reduce the survival rate and scratch healing rate of PANC-1 cells,reduce the number of invasive cells,up-regulate miR-339-5p expression and down-regulate HSPA8 mRNA and protein levels(P＜0.05),while these effects were blocked by down-regulated miR-339-5p(P＜0.05).In addition,HSPA8 was the target gene of miR-339-5p,and knockdown of HSPA8 reversed the regulatory effect of Isov on the malignant biological behavior of PANC-1 cells.In vivo studies confirmed that after Isov treatment,the tumor volume and weight of nude mice decreased,the expression of miR-339-5p was increased and the expression of HSPA8 mRNA was decreased(P＜0.05).Conclusion Isov may inhibit the proliferation,migration and invasion of PC cells through the miR-339-5p/HSPA8 axis.

The SVA and different serotypes of VSV(VSNJV and VSIV)are susceptible to infect pigs and cause blister injuries to the lips and hoof of pigs.The clinical symptoms of diseases caused by these viruses are very similar,which is easy to cause misdiagnosis.Therefore,a multiplex nano-PCR method was developed for the simultaneous defection of VSV,VSNJV and VSIV.In this stud-y,three pairs of specific primers were designed according to the SVA-P gene,VSNJV-N gene and VSIV-N gene.The optimal annealing temperature and optimal primer concentration were tested,and the reaction system and conditions were optimized.We have developed a novel,rapid and sensitive multiple nano-PCR detection method for simultaneous detection of SVA,VSNJV and VSIV,which was developed by using nano-metal materials.The specific test results showed that the method could specifically amplify the target genes of SVA,VSNJV and VSIV,with no cross-reactivity to PRV,ASFV,PCV2 and PHEV.The sensitivity test results showed that the minimum nucleic acid detection of the method was 10 copies/μL,which sensitivity was great.In addition,the optimal primers showed good reactivity and stability to different batches of enzymes and plasmids.There were 7 among 50 of diseased pig samples were SVA positive by multiple nano-PCR detec-tion method,and 5 out of 50 of diseased pig samples were SVA positive by ordinary single PCR method.Moreover,no VSNJV and VSIV were detected by the two methods.In conclusion,this es-tablished multiple nano-PCR detection method has higher specificity and sensitivity in the detec-tion of SVA,VSNJV and VSIV.And this study could provide technical support for the rapid differ-ential diagnosis,prevention and control of swine viral vesicular diseases in clinical settings.

Objective To investigate the effect of methamphetamine(METH)on production of cytokines in RAW264.7 cells stimulated by lipopolysaccharide(LPS)and to explore the possible mechanism.Methods To study the effect of METH on the production of cytokines by LPS-stimulated RAW264.7 cells,the cells were divided into five groups:blank control group,LPS stimulation group,and three METH intervention groups at different concentrations(20,100,and 200 μmol/L),which were METH/LPS groups.To study the role of the dopamine D3 receptor(D3R)in the process,the D3R antagonist NGB2904 was used,and there were five groups:blank control group,LPS stimulation group,METH/LPS group,NGB2904/LPS group,and NGB2904/METH/LPS group.Cell viability was determined using the MTT assay.The levels of IL-6,IL-10,and TNF-α in the supernatants were determined using ELISA.The expressions of p-ERK1/2 and ERK1/2 were determined by Western blotting.Results Compared with the blank control group,the LPS stimulation group and the METH/LPS group had significantly increased cell viability,IL-6,IL-10,TNF-α,and ERK1/2 phosphorylation(P＜0.05).Compared with the LPS stimulation group,the METH/LPS group showed no obvious change in cell viability(P＞0.05),but significantly decreased levels of IL-6,IL-10,and ERK1/2 phosphorylation(P＜0.05).The inhibitory effect of METH showed a dose-dependent manner,with the highest inhibitory effect at METH 200 μmol/L.Furthermore,after the D3R antagonist was used,the results showed that compared with the LPS stimulation group,the cell activity was not significantly changed(P＞0.05),but the levels of IL-6,IL-10 and ERK1/2 phosphorylation were significantly decreased in the METH/LPS group and the NGB2904/LPS group(P＜0.05).Compared with the METH/LPS group or the NGB2904/LPS group,the levels of IL-6,IL-10 and ERK1/2 phosphorylation were further decreased in the NGB2904/METH/LPS group(P＜0.05).Conclusion METH might inhibit the phosphorylation of ERK1/2,thereby reducing the production of IL-6 and IL-10 in RAW264.7 cells stimulated by LPS.The immunosuppressive effect of METH is similar to the immune effect when D3R is antagonized.

Objective To investigate the effect of methamphetamine(METH)on production of cytokines in RAW264.7 cells stimulated by lipopolysaccharide(LPS)and to explore the possible mechanism.Methods To study the effect of METH on the production of cytokines by LPS-stimulated RAW264.7 cells,the cells were divided into five groups:blank control group,LPS stimulation group,and three METH intervention groups at different concentrations(20,100,and 200 μmol/L),which were METH/LPS groups.To study the role of the dopamine D3 receptor(D3R)in the process,the D3R antagonist NGB2904 was used,and there were five groups:blank control group,LPS stimulation group,METH/LPS group,NGB2904/LPS group,and NGB2904/METH/LPS group.Cell viability was determined using the MTT assay.The levels of IL-6,IL-10,and TNF-α in the supernatants were determined using ELISA.The expressions of p-ERK1/2 and ERK1/2 were determined by Western blotting.Results Compared with the blank control group,the LPS stimulation group and the METH/LPS group had significantly increased cell viability,IL-6,IL-10,TNF-α,and ERK1/2 phosphorylation(P＜0.05).Compared with the LPS stimulation group,the METH/LPS group showed no obvious change in cell viability(P＞0.05),but significantly decreased levels of IL-6,IL-10,and ERK1/2 phosphorylation(P＜0.05).The inhibitory effect of METH showed a dose-dependent manner,with the highest inhibitory effect at METH 200 μmol/L.Furthermore,after the D3R antagonist was used,the results showed that compared with the LPS stimulation group,the cell activity was not significantly changed(P＞0.05),but the levels of IL-6,IL-10 and ERK1/2 phosphorylation were significantly decreased in the METH/LPS group and the NGB2904/LPS group(P＜0.05).Compared with the METH/LPS group or the NGB2904/LPS group,the levels of IL-6,IL-10 and ERK1/2 phosphorylation were further decreased in the NGB2904/METH/LPS group(P＜0.05).Conclusion METH might inhibit the phosphorylation of ERK1/2,thereby reducing the production of IL-6 and IL-10 in RAW264.7 cells stimulated by LPS.The immunosuppressive effect of METH is similar to the immune effect when D3R is antagonized.

Objective To explore the biological behavior and mechanism of isovitexin(Isov)on pancreatic cancer cells.Methods Isov was used to treat the human normal pancreatic ductal epithelial cells HPDE and PC cell lines,and CCK-8 was used to detect the cell proliferation and calculate the half inhibitory concentration(IC50).The PC cell line PANC-1 cells were grouped into the control group,the Isov group,the Isov+in-miR-NC group,the Isov+in-miR-339-5p group,the Isov+in-miR-339-5p+si-NC group and the Isov+in-miR-339-5p+si-HSPA8 group.The survival,migration and invasion of PANC-1 cells were detected by CCK-8,scratch healing assay and Transwell assay.Real time fluorescence quantitative PCR was used to detect the mRNA expression of miR-339-5p and heat shock protein family A member 8(HSPA8)in PANC-1 cells.Western blot assay was used to detect protein HSPA8 expression in various groups of cells.Dual luciferase reporter gene was used to detect the targeting effect of miR-339-5p and HSPA8.A xenograft nude mouse model was used to determine the in vivo anticancer effects of Isov.Results Isov inhibited PC cell proliferation but had little cytotoxicity to HPDE cells.Isov could obviously reduce the survival rate and scratch healing rate of PANC-1 cells,reduce the number of invasive cells,up-regulate miR-339-5p expression and down-regulate HSPA8 mRNA and protein levels(P＜0.05),while these effects were blocked by down-regulated miR-339-5p(P＜0.05).In addition,HSPA8 was the target gene of miR-339-5p,and knockdown of HSPA8 reversed the regulatory effect of Isov on the malignant biological behavior of PANC-1 cells.In vivo studies confirmed that after Isov treatment,the tumor volume and weight of nude mice decreased,the expression of miR-339-5p was increased and the expression of HSPA8 mRNA was decreased(P＜0.05).Conclusion Isov may inhibit the proliferation,migration and invasion of PC cells through the miR-339-5p/HSPA8 axis.

The SVA and different serotypes of VSV(VSNJV and VSIV)are susceptible to infect pigs and cause blister injuries to the lips and hoof of pigs.The clinical symptoms of diseases caused by these viruses are very similar,which is easy to cause misdiagnosis.Therefore,a multiplex nano-PCR method was developed for the simultaneous defection of VSV,VSNJV and VSIV.In this stud-y,three pairs of specific primers were designed according to the SVA-P gene,VSNJV-N gene and VSIV-N gene.The optimal annealing temperature and optimal primer concentration were tested,and the reaction system and conditions were optimized.We have developed a novel,rapid and sensitive multiple nano-PCR detection method for simultaneous detection of SVA,VSNJV and VSIV,which was developed by using nano-metal materials.The specific test results showed that the method could specifically amplify the target genes of SVA,VSNJV and VSIV,with no cross-reactivity to PRV,ASFV,PCV2 and PHEV.The sensitivity test results showed that the minimum nucleic acid detection of the method was 10 copies/μL,which sensitivity was great.In addition,the optimal primers showed good reactivity and stability to different batches of enzymes and plasmids.There were 7 among 50 of diseased pig samples were SVA positive by multiple nano-PCR detec-tion method,and 5 out of 50 of diseased pig samples were SVA positive by ordinary single PCR method.Moreover,no VSNJV and VSIV were detected by the two methods.In conclusion,this es-tablished multiple nano-PCR detection method has higher specificity and sensitivity in the detec-tion of SVA,VSNJV and VSIV.And this study could provide technical support for the rapid differ-ential diagnosis,prevention and control of swine viral vesicular diseases in clinical settings.

Objective:To compare iodine nutritional status of different populations consuming iodized salt and non-iodized salt in iodine adequate areas, and to provide a basis for formulating iodine supplementation strategies.Methods:In October 2021, Luyi County in Henan Province was selected as an iodine adequate area consuming iodized salt, while Ningling County was selected as an iodine adequate area consuming non-iodized salt. Stratified by water iodine (50 - 59, 60 - 69, 70 - 79, 80 - 89, 90 - 100 μg/L), one village was selected from each layer. One hundred children aged 8 - 10, one hundred adults, and 20 pregnant women were selected from each village to collect their urine and salt samples for testing salt and urinary iodine, and their thyroid gland was measured by ultrasound.Results:A total of 600 salt samples in Luyi County were collected, with the coverage rate of iodized salt (99.8%, 599/600) and the consuming rate of qualified iodized salt (95.5%, 573/600). A total of 1 008 salt samples in Ningling County were collected, with the rate of non-iodized salt (93.8%, 946/1 008). The median urinary iodine of children in Luyi County ( n = 240) was higher than that in Ningling County ( n = 468, 305.0 vs 232.0 μg/L, Z = - 8.10, P < 0.001). There was no statistically significant difference in median urinary iodine between pregnant women in Luyi County ( n = 120) and Ningling County ( n = 53, 240.0 vs 236.0 μg/L, Z = - 1.02, P = 0.306). The median urinary iodine of adults in Luyi County ( n = 238) was higher than that in Ningling County ( n = 486, 289.0 vs 178.5 μg/L, Z = - 11.14, P < 0.001). Children's urinary iodine ( r s = 0.21, P = 0.001) in Luyi County and adults' urinary iodine ( r s = 0.17, P < 0001) in Ningling County were positively correlated with water iodine. There was no statistically significant difference in the incidence of thyroid enlargement in children between Luyi County (0.8%, 2/240) and Ningling County (0.4%, 2/468, χ 2 = 0.80, P = 0.586), but the incidence of thyroid nodules in children in Luyi County (11.2%, 27/240) was higher than that in Ningling County (1.7%, 8/468, χ 2 = 27.36, P < 0.001). The incidence of thyroid nodules in pregnant women in Luyi County (23.3%, 28/120) was lower than that in Ningling County (46.5%, 33/71, χ 2 = 10.99, P = 0.001). There was no statistically significant difference in the incidence of adult thyroid nodules between Luyi County and Ningling County (χ 2 = 0.86, P = 0.354), with a ratio of 29.6% (71/240) to 32.9% (160/486). Conclusions:Providing population with non-iodized salt in iodine adequate areas, the overall iodine nutrition is at an appropriate level. However, children consuming iodized salt in iodine adequate areas have high level of iodine nutrition, and it is necessary to consider supplying non-iodized salt or reducing the concentration of iodized salt. Pregnant women in iodine adequate area should maintain the current policy of supplying iodized salt unchanged.

Objective: To evaluate the changes in the endoplasmic reticulum stress in the spinal cord of rats with diabetic neuropathic pain (DNP). Methods: Clean-grade healthy male Sprague-Dawley rats, aged 8 weeks, weighing 120-160 g, were fed a high-fat and high-glucose diet for 8 weeks, then diabetes mellitus was induced by intraperitoneal streptozotocin 35 mg/kg and confirmed by blood glucose level >16.7 mmol/L 3 days later.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at day 14 after injection.The establishment of DNP model was considered successful when MWT and TWL were lower than 85% of the baseline value.Fifteen rats in which the DNP model was successfully established served as DNP group, 15 rats in which the DNP model was not successfully established served as non-NDNP group (NDNP), and another 15 normal rats were selected and served as control group (group C). The MWT and TWL were measured at 3, 7 and 14 days after successful establishment of the model.The rats were then sacrificed, and the lumbar enlargement segments (L_4-6) of the spinal cord were harvested to detect the expression of inositol-requiring enzyme-1α, phosphorylated JNK (p-JNK) and Beclin1 by Western blot. Results: Compared with C and NDNP groups, the MWT was significantly decreased and the TWL was shortened at 3, 7 and 14 days after successful establishment of the model, and the expression of inositol-requiring enzyme-1α, p-JNK and Beclin1 was up-regulated in DNP group (P<0.05). Conclusion The mechanism of development and maintenance of DNP may be related to aggravated endoplasmic reticulum stress in the spinal cord and to autophagy induced by up-regulated expression of p-JNK in rats.

Objective To determine whether there is correlation between intracranial and extracranial arterial stenosis in patients with ischemic stroke,and to understand the difference of the main risk factors between them.Methods One hundred and sixty-eight patients with symptomatic ischemic stroke were selected in order to analyze the correlation and risk factors of intracranial arterial stenosis and extracranial arterial stenosis.Results In the 168 cases,123 patients(73.2%)were with abnormal artery(include intimal thickening)and 100 patients(59.5%)were diagnosed with artery stenosis,including 33 cases of intracranial artery stenosis,48 cases of extracranial artery stenosis and 19 cases of intracranial and extracranial stenosis.The rate of extracranial artery stenosis(39.9%)was higher than that of intracranial artery stenosis(31.0%),but there was no significant difference between them(χ2 =2.93,P=0.11).There was no definite correlation between them(χ2 =0.35,P=0.61)and there were statistically significant differences in age and arterial pressure between patients with intracranial and mean extracranial artery stenosis(t=1.98,P=0.05;t=5.42,P<0.001),but the gender,blood pressure,blood glucose and dyslipidemia rate showed no significant difference (χ2 =1.15,3.41,0.43,0.81,P>0.05).Conclusion Extracranial arterial stenosis may not predict the possibility of intracranial arterial stenosis,and extracranial arterial stenosis may be more likely to cause ischemic stroke.

Objective: To investigate the safety and efficacy of transthoracic minimally invasive patent ductus arteriosus (PDA) occlusion in infants and young children.
Methods: We retrospectively analyzed 105 infants and young children who received the transthoracic minimally invasive PDA occlusion in our hospital from 2012-10 to 2014-10. According to PDA diameter, patients were divided into 2 groups:Group A, the patients with PDA diameter ≥ 4 mm,n=64 and group B, the patients with 2 mm ≤ PDA diameter < 4 mm,n=41. All patients received the left third parasternal intercostal incision under suprasternal echocardiography guidance. The operative effect was evaluated by transthoracic echocardiography, and the follow-up study was performed at 1 month, 3 months, 6 months period and then annually after the operation by echocardiography.
Results: All 105 patients had successfully implanted PDA occluders. The patients’ gender, age, body weight, tracheal intubation time and the in-hospital time were similar between 2 groups,P>0.05. Compared with Group B, Group A had the larger diameters of PDA (5.7 ± 1.4) mm vs (2.7 ± 0.6) mm, P<0.001, PDA occluders (10.6 ± 1.8) mm vs (7.2 ± 1.3) mm, P<0.001, and the higher rates of moderate and severe post-operative thrombocytopenia 10.9% (7/64) vs 0% (0/41),P=0.028, immediate post-operative residual shunt as 15.6% (10/64) vs 2.4% (1/41),P=0.031. There was 1 patient in Group A suffered from pericardial tamponade due to hemorrhage at 2 days after operation and he was cured by emergent pericardial drainage. The patients were followed-up for (11.6 ± 7.8) months. The 1 month post-operative residual shunt was similar between 2 groups as 1.6% (1/64 ) vs 0% (0/41),P=0.421, and there was no residual shunt at 3 months after the operation. There were no complications of occluder detachment, hemolysis, pericardial effusion, left pulmonary artery or descending aortic stenosis occurred during the follow-up period.
Conclusion: Transthoracic minimally invasive PDA occlusion is a safe and effective method to treat the relevant infants and young children, while the post-operative residual shunt and thrombocytopenia should be closely observed in patients with large PDA.