Nocardia is a Gram-positive pathogen responsible for causing dairy mastitis,which leads to purulent granulomatous lesions in mammary tissue and can significantly impact the dairy indus-try,resulting in substantial economic losses.To develop a rapid and accurate diagnostic method for detecting Nocardia of bovine origin,a conserved sequence of the 16S rRNA gene from Nocardia was selected from the NCBI database.Based on this sequence,a pair of primers and a TaqMan fluo-rescent quantitative probe were designed.The validation of the TaqMan fluorescence quantitative PCR(qPCR)method found in this study showed that the Ctvalue had a good linear relationship with recombinant plasmid concentrations ranging from 1×1010 to 1×102 copies/μL,with a regres-sion equation of y=-3.536x+43.78,a correlation coefficient(R2)of 0.997 5,a slope of-3.536,and an amplification efficiency(E)of 91％(where 90％＜E＜110％).The specificity was strong,with no cross-reactions with other pathogens.The standard curve had a high sensitivity with a low-er detection limit of 1 × 102 copies/μL,it was 100-fold higher than conventional PCR.The repeatability of the standard curve was also good.Both intra-and inter-group coefficients of varia-tion were below 2％.Using this method,234 milk samples and 80 environmental samples were tested using this method,respectively,with a positive detection rate of 27.07％,whereas conven-tional PCR had a positive detection rate of 19.43％,indicating that this method was more sensitive compared to conventional PCR.The fluorescent quantitative PCR detection method established in this study provides an effective means for the clinical detection of Nocardia in dairy cows.

Objective To compare the complications,etiology,and clinical manifestations between pediatric patients with heart valve disease undergoing surgical treatment.Methods A retrospective analysis was conducted on the clinical data of 169 pediatric patients aged 0-14 years diagnosed with heart valve abnormalities at Wuhan Union Hospital from January 2019 to October 2022.Patients were divided into two groups based on whether they received surgical intervention for valve abnormali-ties:the surgical treatment group and the medical treatment group.Results There were significant differences in age,type and grade of heart failure,club-finger,pneumonia and cough,cardiac morphology,arrhythmia,ventricular septal width and left ven-tricular ejection fraction(all P＜0.05).Univariate and multivariate binary logistic regression analyses revealed that left ventricu-lar ejection fraction(OR=1.206,95％CI:1.072-1.357,P＜0.05)was an independent risk factor for the surgical treatment group.Conclusion The clinical manifestations of heart valve abnormalities in children are often atypical.Clinicians should ac-tively evaluate the etiology based on clinical symptoms and echocardiographic findings,and develop individualized treatment strategies,either surgical or medical,to prevent the progression of severe heart disease.

Nocardia is a Gram-positive pathogen responsible for causing dairy mastitis,which leads to purulent granulomatous lesions in mammary tissue and can significantly impact the dairy indus-try,resulting in substantial economic losses.To develop a rapid and accurate diagnostic method for detecting Nocardia of bovine origin,a conserved sequence of the 16S rRNA gene from Nocardia was selected from the NCBI database.Based on this sequence,a pair of primers and a TaqMan fluo-rescent quantitative probe were designed.The validation of the TaqMan fluorescence quantitative PCR(qPCR)method found in this study showed that the Ctvalue had a good linear relationship with recombinant plasmid concentrations ranging from 1×1010 to 1×102 copies/μL,with a regres-sion equation of y=-3.536x+43.78,a correlation coefficient(R2)of 0.997 5,a slope of-3.536,and an amplification efficiency(E)of 91％(where 90％＜E＜110％).The specificity was strong,with no cross-reactions with other pathogens.The standard curve had a high sensitivity with a low-er detection limit of 1 × 102 copies/μL,it was 100-fold higher than conventional PCR.The repeatability of the standard curve was also good.Both intra-and inter-group coefficients of varia-tion were below 2％.Using this method,234 milk samples and 80 environmental samples were tested using this method,respectively,with a positive detection rate of 27.07％,whereas conven-tional PCR had a positive detection rate of 19.43％,indicating that this method was more sensitive compared to conventional PCR.The fluorescent quantitative PCR detection method established in this study provides an effective means for the clinical detection of Nocardia in dairy cows.

Objective To compare the complications,etiology,and clinical manifestations between pediatric patients with heart valve disease undergoing surgical treatment.Methods A retrospective analysis was conducted on the clinical data of 169 pediatric patients aged 0-14 years diagnosed with heart valve abnormalities at Wuhan Union Hospital from January 2019 to October 2022.Patients were divided into two groups based on whether they received surgical intervention for valve abnormali-ties:the surgical treatment group and the medical treatment group.Results There were significant differences in age,type and grade of heart failure,club-finger,pneumonia and cough,cardiac morphology,arrhythmia,ventricular septal width and left ven-tricular ejection fraction(all P＜0.05).Univariate and multivariate binary logistic regression analyses revealed that left ventricu-lar ejection fraction(OR=1.206,95％CI:1.072-1.357,P＜0.05)was an independent risk factor for the surgical treatment group.Conclusion The clinical manifestations of heart valve abnormalities in children are often atypical.Clinicians should ac-tively evaluate the etiology based on clinical symptoms and echocardiographic findings,and develop individualized treatment strategies,either surgical or medical,to prevent the progression of severe heart disease.

Objective:To investigate the regulatory effect of WNT1-inducible signaling pathway protein 2(WISP2)on macrophage polarization in palmitic acid(PA)and lipopolysaccharide(LPS)-induced inflammation.Methods:The macrophage cell line RAW264.7 was treated with different concentrations of WISP2 protein, and cell viability was determined by means of luminescence assay using Cell-Titer Glo to determine the concentration of WISP2.The cells were divided into control group, palmitic acid group, palmitic acid combined with different concentrations of WISP2 group(10 μg/L and 100 μg/L)and lipopolysaccharide group, lipopolysaccharide combined with different concentrations of WISP2 group(10 μg/L and 100 μg/L). mRNA expression of M1 and M2 macrophages phenotype of each group were detected by real-time quantitative polymerase chain reaction.The protein expression of important inflammatory factors, TNF-α and IL-6, were evaluated by ELISA.Results:Compared with the control group, both 10 μg/L and 100 μg/L WISP2 groups had no effect on the activity of RAW264.7 cells, but significantly up-regulated the expression of various inflammatory factors, including Tnfα(1.877±0.039, 2.202±0.034, F=309.7, P<0.001), Il6(1.418±0.056, 1.506±0.059, F=81.39, P<0.001), Mcp1(1.620±0.014, 1.982±0.125, F=71.45, P<0.001), Ccl3(1.892±0.118, 1.942±0.132, F=32.93, P<0.001), and iNos(1.691±0.201, 1.548±0.090, F=13.60, P<0.05). mRNA in macrophages, and significantly down-regulated the expression of anti-inflammatory factors, including Tgfβ(1.376±0.025, 2.152±0.107, F=1.846, P<0.05), CD206(2.123±0.031, 3.139±1.663, F=8.037, P<0.05), Il4(2.098±0.464, 2.494±0.141, F=48.68, P<0.01), and Il10(1.303±0.216, 1.574±0.274, F=5.774, P<0.05)mRNA, causing M1 type macrophage polarization.Compared with the control group, 100 μmol/L palmitic acid could mildly but significantly increase the expression of inflammatory factors such as TNF-α and IL-6 at the transcriptional and protein levels.Compared with palmitic acid stimulation alone, the combination of palmitic acid and WISP2 further promoted the protein expression of macrophage inflammatory factors TNF-α[(589.4±17.0)ng/L, (692.6±83.4)ng/L, F=56.38, P<0.05], IL-6[(15.13±1.14)ng/L, (13.33±1.22)ng/L, F=23.32, P<0.001]and the mRNA expression of chemokines Mcp1(160±9.796, 140±18.91, F=141.1, P<0.0001)and C cl3(17.76±1.92, 14.41±1.27, F=125.2, P<0.0001). Compared with the control group, 100 μg/L lipopolysaccharide strongly stimulated the expression of inflammatory factors such as TNF-α[(3444±423)ng/L, F=71.20, P<0.0001]and IL-6[(497.0±41.2)ng/L, F=63.50, P<0.0001]in macrophages at the protein level.Compared with lipopolysaccharide stimulation alone, the combination of lipopolysaccharide and WISP2 further significantly up-regulated the mRNA expression of chemokines Mcp1(106.8±8.7, 118.7±4.6, F=251.5, P<0.0001)and Ccl3(35.3±12.5, 116.4±4.5, F=160.1, P<0.0001). Conclusions:The adipokine WISP2 can promote M1 macrophage polarization in palmitic acid and lipopolysaccharide-induced inflammation, and it had distinct regulation in macrophage polarization under different inflammatory response conditions.

Objective: To describe the prevalence of overweight and obesity among children and adolescents in Jinan City according the national and international criteria, as well as to compare the Chinese and international standards, so as to provide basic data for relevant intervention strategies. Methods: From September to December 2021, using the height and weight data obtained from the health checkup reports of 746 985 primary and secondary school students in Jinan City in 2021, body mass index(BMI) was calculated to evaluate the current status of the prevalence of overweight and obesity among this sample of children and adolescents aged 6- 18 years old. The consistency of the Chinese standard, U.S. Centers for Disease Control and Prevention (CDC) standard, International Obesity Task Force (IOTF) standard, and World Health Organization (WHO) standard was compared by calculating the weighted Kappa value. Results: Based on the Chinese, U.S. CDC, IOTF and WHO standards, the overweight rates of children and adolescents aged 6-18 years in Jinan were 17.3%, 16.5%, 21.1% and 13.6%, respectively, and the obesity rates were 23.1%, 19.5%, 13.5% and 6.6%, respectively. The overweight and obesity rates of boys were higher than those of girls under the four criteria (China: χ 2=1 418.48, 9 868.51, U.S. CDC: χ 2=145.78, 23 211.41, IOTF: χ 2=1 326.94, 13 615.62, WHO: χ 2=873.13, 46.41, P <0.01).The overweight rate of adolescents in the 13-18 year-old age group was higher than that of children aged 6-12 years (China: χ 2=29.13, U.S. CDC: χ 2=6.43, IOTF: χ 2=15.87, WHO: χ 2=19.48, P <0.01) and the obesity rate of the group aged 6-10 years was significantly higher than that of the group aged 11-18 years (China: χ 2=217.02, U.S. CDC: χ 2=227.54, IOTF: χ 2=171.91, WHO: χ 2=165.91, P <0.01). The rates of overweight and obesity in urban children and adolescents were slightly higher than those in rural children and adolescents (China: χ 2=25.06, 245.12, U.S. CDC: χ 2=56.13, 205.93, IOTF: χ 2= 182.34 , 22.04, WHO: χ 2=200.88, 21.61, P <0.01). The Chinese standard showed good agreement with the IOTF standard and U.S. CDC standard, with weighted Kappa values of 0.72 and 0.83, but poor agreement with the WHO standard, with a weighted Kappa value of 0.33. Conclusion The current prevalence of overweight and obesity among children and adolescents in Jinan is serious. The Chinese standard has good consistency with IOTF standard and American CDC standard, and poor consistency with WHO standard.

ObjectiveTo explore the effects and possible mechanism of Wenshen Tongdu Formula （温肾通督方， WTF） on spinal cord injury. MethodsThirty-six C57BL/6 female mice were randomly divided into sham operation group， model group and WTF group， with 12 mice in each group. The spinal cord injury model was established in the model group and the WTF group using the modified Allen's method， while in the sham operation group the spinal cord was only exposed. Since the 1st day after surgery， 50 g/（kg·d） of WTF solution was given to the WTF group by gavage， while 20 ml/（kg·d） of normal saline was given to the sham operation and model group by gavage， all for 14 days. Before surgery and on the 1st， 7th， and 14th days after surgery， the motor function of the mice was evaluated using the inclined plane test and hind limb motor function score （by BMS）. On the 3rd day after surgery， the nerve electrophy-siology was detected through electromyography and motor evoked potential； the spleen length was measured， and B cells in the spleen were sorted by magnetic beads； the differential expression of proteins were detected through proteomics technology； and the protein expression of mitochondrial outer membrane transport porin 20 （Tom20） and downstream cleaved caspase-3 in spleen B cells were measured using Western blotting. On the 14th day after surgery， MRI was used to observe the recovery of the spinal cord. ResultsCompared to those in the sham operation group at the same time， the BMS scores and subscores and the inclined plane test angle in the model group were reduced on the 1st， 7th and 14th days after surgery； the peak value of electromyogram and motor evoked potential were reduced， and the spleen length was shortened， while the expression of Tom20 and cleaved caspase-3 increased in splenic B cells increased （P＜0.05）. Compared to those in the model group at the same time， the BMS subscores on the 14th day and the angle of the inclined plane test on the 7th and 14th days after surgery increased in the WTF group； the peak value of electromyography and motor evoked potential， as well as the length of spleen increased， and the expression of Tom20 and cleaved caspase-3 decreased （P＜0.05）. The proteomics results showed that there were 100 differential proteins in the WTF group versus the model group， of which 37 were up-regulated and 63 were down-regulated. GO enrichment analysis showed that differential proteins mainly played their roles in oxygen binding， exogenous apoptosis negative feedback， zinc ion response， and oxygen transport. KEGG enrichment analysis showed that differential proteins were mainly concentrated in metabolic pathways， Huntington's disease， oxidative phosphorylation and other pathways. Subcellular localization showed that differential proteins were associated with mitochondria. Magnetic resonance imaging on the 14th day after surgery showed that the spinal cord structure of the mice in the sham operation group was intact， and the segments were clear， with normal spinal cord signal； the low signal area in the spinal cord injury area increased in the model group， and the spinal cord became significantly thinner； the injured segment had obvious depression in the WTF group， but the structure was more complete than that in the model group. ConclusionWTF may promote spinal cord injury repair by regulating immune function， and its mechanism may be related to inhibiting pyroptosis of spleen B cells.

Detection and identification standard of hookworm—Hookworm larvae coproculture techniques (WS/T 791—2021) is the first recommended technical standard for hookworm detection and species identification using the hookworm larvae coproculture technique in China. This standard was issued on November 23, 2021, and had been in effect since May 1, 2022. This article provides a detailed interpretation pertaining to the background, drafting process, main contents, and dos and don’ts for better understanding and application of this standard among professionals working in disease control and prevention institutions and medical institutions.

Objective:To investigate the effects of neonatal respiratory distress syndrome(NRDS)on thymus of premature infants.Methods:We collected baseline data from premature infants with gestational age of 28~32 weeks in neonatal intensive care unit of Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 1, 2019 to December 31, 2019.The largest transverse diameter and the sagittal of thymus were measured by ultrasonography within 24 h of birth, then, the thymic index(TI)and thymic weight index(TWI)were calculated to assess the size of thymus.The preterm neonates were divided into NRDS group and non-NRDS group according to the diagnosic criteria of NRDS, and the two groups were then divided into antenatal corticosteroid administration(ACS)group and non-ACS group according to ACS exposure.We then compared the TI and TWI between these groups.Results:One hundred and sixty-three preterm neonates were enrolled in our study, including 98 NRDS preterm neonates and 65 non NRDS preterm neonates.After matching gestational age and birth weight of the preterm neonates from two groups, 65 preterm neonates with NRDS comprised the NRDS group, and 65 preterm neonates without NRDS served as controls.Preterm neonates in NRDS group had significantly smaller TI[(1.788 ± 0.803)cm 3 vs.(2.420±1.068)cm 3, t=3.818, P<0.01] and TWI[(1.278 ± 0.380)cm 3/kg vs.(1.695 ± 0.491)cm 3/kg, t=5.401, P<0.01] than those in non-NRDS group.Besides, preterm neonates in NRDS group had smaller lymphocytes count[(3.729 ± 1.263)×10 9/L vs.(4.437 ± 1.608)×10 9/L, t=2.789, P<0.01] than that in non-NRDS group.For NRDS preterm neonates, TI[(1.487 ± 0.515)cm 3 vs(2.185 ± 0.942)cm 3, t=3.542, P<0.01] ]and TWI[(1.134± 0.311)cm 3/kg vs(1.469± 0.385)cm 3/kg, t=3.882, P<0.01] in ACS group were significantly smaller than those in non-ACS group.For non-NRDS preterm neonates, TI and TWI in ACS group also were significantly smaller than those in non-ACS group( t=2.676、3.659, P<0.05). Conclusion:NRDS is associated with thymic involution of preterm neonates, and ACS exposure affected the size of thymic in premature infants.

Objective:To establish an in vivo model of acute radiation esophagitis in Wistar rats based on a small animal precision radiotherapy platform (SARRP). Methods:Thirty-six Wistar rats were randomly divided into control group, 40, 60 and 75 Gy groups. Based on MRI images, the esophageal target area of rats was outlined and the radiotherapy plan was formulated. The rats were respectively irradiated with 0, 8, 12 and 15 Gy per day for 5 consecutive days. The changes of body weight, food intake, esophageal pathology and magnetic resonance imaging were observed.Results:The body weight of rats in 75 Gy group decreased significantly on the 6th day after irradiation (IR) ( P<0.05). The esophageal tissue of rats in each IR group was thicker than that in control ( F = 14.20, P < 0.05). HE staining showed that the formation rate of radiation-induced esophagitis in 40 Gy and 60 Gy groups were 4/5 and 5/5, respectively, mainly mild. In 75 Gy group, the incidence of radiation-induced esophagitis approached to 5/5, of which 3/5 was severe at 9 d post-IR. The pathological injury scores [ M( Q1, Q3)] of rats in each group were 0, 1.0 (0.5, 2.5), 1.0 (1.0, 2.5) and 4.0 (1.5, 6.0) on the 9th day after IR. There was significant difference between the 75 Gy group and the control group ( H=12.69, P<0.05). After dynamic monitoring of neck MRI images, it was found that the esophageal signal of rats in each IR group increased and widened at 9 d post-irradiation. Conclusions:The animal model of acute radiation-induced esophagitis in rats was successfully established based on a small animal precision radiotherapy platform combined with MRI. 75 Gy was the best irradiation dose and the 9th day was the best observation time point.