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 investigate the relationship between calcium-sensing receptor(CaSR)gene rs17251221,rs60388563 loci polymorphisms and the genetic susceptibility of breast cancer(BC).Methods A total of 122 patients with breast cancer admitted to Hengshui Second People's Hospital from January 2022 to June 2024 were selected as the BC group,and 100 healthy women without blood relationship at the same time were selected as the control group.The polymorphisms of CaSR gene rs17251221,rs60388563 loci were detected by polymerase chain reaction(PCR).The clinical data,genotype distribution and allele frequency were compared between the two groups.Non-conditional Logistic regression model was used to analyze the correlation between CaSR gene polymorphism and genetic susceptibility of BC patients.Results Comparing the general data of the two group,the proportion of family history of cancer in the BC group was significantly higher than that in the control group,and the difference was statistically significant(t=12.246,P<0.05).The genotype distribution of CaSR gene rs17251221 and rs60388563 loci in the control group and the BC group was consistent with the Hardy-Weinberg equilibrium law(χ2=0.087～1.202,P＞0.05),which was representative of the population.Under the co-dominant model of rs17251221 and rs60388563 loci,the risk of BC in GG genotype was significantly increased(OR=1.493,95%CI=1.070～2.108;OR=1.483,95%CI=1.034～2.121).Under the dominant model(AA vs GA+GG)and recessive model(GG vs GA+AA),the rs17251221 locus carrying G allele(OR=1.371,95%CI=1.023～1.824)and A allele(OR=0.524,95%CI=0.221～0.926)was significantly associated with the risk of BC(P<0.05).Under the dominant model(CC vs GC+GG)and recessive model(GG vs GC+CC),the rs60388563 locus carrying G allele(OR=1.245,95%CI=1.107～1.461)and C allele(OR=0.682,95%CI=0.523～0.974)was significantly associated with the risk of BC(P<0.05).Conclusion The rs17251221,rs60388563 loci polymorphisms of CaSR gene are closely related to BC susceptibility,and the risk of BC is high in individuals carrying G allele.

Male infertility has become a global concern, accounting for 20-70% of infertility. Dysfunctional spermatogenesis is the most common cause of male infertility; thus, treating abnormal spermatogenesis may improve male infertility and has attracted the attention of the medical community. Mitochondria are essential organelles that maintain cell homeostasis and normal physiological functions in various ways, such as mitochondrial oxidative phosphorylation (OXPHOS). Mitochondrial OXPHOS transmits electrons through the respiratory chain, synthesizes adenosine triphosphate (ATP), and produces reactive oxygen species (ROS). These mechanisms are vital for spermatogenesis, especially to maintain the normal function of testicular Sertoli cells and germ cells. The disruption of mitochondrial OXPHOS caused by external factors can result in inadequate cellular energy supply, oxidative stress, apoptosis, or ferroptosis, all inhibiting spermatogenesis and damaging the male reproductive system, leading to male infertility. This article summarizes the latest pathological mechanism of mitochondrial OXPHOS disorder in testicular Sertoli cells and germ cells, which disrupts spermatogenesis and results in male infertility. In addition, we also briefly outline the current treatment of spermatogenic malfunction caused by mitochondrial OXPHOS disorders. However, relevant treatments have not been fully elucidated. Therefore, targeting mitochondrial OXPHOS disorders in Sertoli cells and germ cells is a research direction worthy of attention. We believe this review will provide new and more accurate ideas for treating male infertility.
Male ; Humans ; Infertility, Male/metabolism* ; Oxidative Phosphorylation ; Mitochondria/metabolism* ; Spermatogenesis/physiology* ; Sertoli Cells/metabolism* ; Oxidative Stress/physiology* ; Animals ; Reactive Oxygen Species/metabolism*

Objective To investigate the relationship between calcium-sensing receptor(CaSR)gene rs17251221,rs60388563 loci polymorphisms and the genetic susceptibility of breast cancer(BC).Methods A total of 122 patients with breast cancer admitted to Hengshui Second People's Hospital from January 2022 to June 2024 were selected as the BC group,and 100 healthy women without blood relationship at the same time were selected as the control group.The polymorphisms of CaSR gene rs17251221,rs60388563 loci were detected by polymerase chain reaction(PCR).The clinical data,genotype distribution and allele frequency were compared between the two groups.Non-conditional Logistic regression model was used to analyze the correlation between CaSR gene polymorphism and genetic susceptibility of BC patients.Results Comparing the general data of the two group,the proportion of family history of cancer in the BC group was significantly higher than that in the control group,and the difference was statistically significant(t=12.246,P<0.05).The genotype distribution of CaSR gene rs17251221 and rs60388563 loci in the control group and the BC group was consistent with the Hardy-Weinberg equilibrium law(χ2=0.087～1.202,P＞0.05),which was representative of the population.Under the co-dominant model of rs17251221 and rs60388563 loci,the risk of BC in GG genotype was significantly increased(OR=1.493,95%CI=1.070～2.108;OR=1.483,95%CI=1.034～2.121).Under the dominant model(AA vs GA+GG)and recessive model(GG vs GA+AA),the rs17251221 locus carrying G allele(OR=1.371,95%CI=1.023～1.824)and A allele(OR=0.524,95%CI=0.221～0.926)was significantly associated with the risk of BC(P<0.05).Under the dominant model(CC vs GC+GG)and recessive model(GG vs GC+CC),the rs60388563 locus carrying G allele(OR=1.245,95%CI=1.107～1.461)and C allele(OR=0.682,95%CI=0.523～0.974)was significantly associated with the risk of BC(P<0.05).Conclusion The rs17251221,rs60388563 loci polymorphisms of CaSR gene are closely related to BC susceptibility,and the risk of BC is high in individuals carrying G allele.

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.

The cartilage-protective effect of ginkgolide C(GC)on the two modeling modalities was investigated based on joint pain,degree of cartilage pathology,ECM degradation process,and level of inflammatory mediator production in rats.Twenty-five SD rats were selected and randomly di-vided into five groups:the control group(Control group),model 1 group(ACLT group),adminis-tration 1 group(ACLT+GC group),model 2 group(MIA group),and administration 2 group(MIA+GC group.)The rats were euthanized after 4 weeks of the test.Femur,tibia and blood samples were collected from the right hind limb of rats.The degree of pathology in the femur and tibia of rats was assessed by saffron O solid green staining and OARSI score.Immunohistochemis-try was used to detect the expression levels of collagen Ⅱ and MMP-13 in cartilage.ELISA was used to detect the changes in the levels of MMP-3,MMP-13,CTX-Ⅱ,COMP,COX-2,INOS,IL-1β,and TNF-α in the serum of rats.Cold sensitivity test and knee extension vocalization test were conducted to detect the degree of joint pain in rats.ACLT could cause more severe structural dam-age to articular cartilage compared with the MIA group.The OARSI scores and the expression of MMP-13 in femur and tibia,and the serum levels of MMP-13,MMP-3,CTX-Ⅱ,and COMP were higher in the ACLT group than those in the MIA group.However,the levels of inflammatory me-diators COX-2,IL-1β,and TNF-α were significantly lower in the ACLT group than in the MIA group(P＜0.0l).GC intervention reduced the OARSI score(P＜0.05 or P＜0.01)and pain scores,inhibited the ECM matrix degrading enzymes(MMP-13,MMP-3),cartilage metabolism markers(CTX-11,COMP),and inflammatory mediators(COX-2,INOS,IL-1β and TNF-α)ex-pression,and promoted collagen Ⅱ synthesis.Both modeling methods resulted in cartilage damage.In particular,the OA model constructed by ACLT+PMMx method in rats had obvious joint dam-age,which was favorable to investigate the degree of cartilage structural damage.GC attenuated cartilage pathological changes,pain severity and inflammatory response in the rat OA model in both groups,thus exerting a cartilage-protective effect.

Objective To investigate the role of histone deacetylase 6(HDAC6)in podocyte injury in diabetic kidney disease(DKD)in mice.Methods 1)The 8-week-old male CD-1 mice were selected to construct the model of DKD with streptozocin(STZ).After the model was established,the mice were intraperitoneally injected with HDAC6 inhibitor CAY10603(5mg/kg/daily)or same volume vehicle as control.The mice were divided into four groups,control(CTL)+vehicle(Veh)(n=5),CLT+CAY10603(n=3),STZ+Veh(n=9),and STZ+CAY10603(n=7).Mice in STZ+Veh and STZ+CAY10603 groups developed DKD,while mice in the CTL+Veh and CTL+CAY10603 groups were served as normal controls.The therapeutic effect was evaluated through urine albumin-to-creatinine ratio(uACR)and renal pathology after the 2-week treatment with CAY10603.2)Human podocytes were cultured in vitro and were divided into four groups as follows:CTL,transforming growth factor-β(TGFβ),TGFβ+CAY10603(250 nmol/L),and TGFβ+CAY10603(500 nmol/L)groups.The control group did not receive any treatment,the last three groups were given 36-h TGFβtreatment at 5 ng/μL,with or without CAY10603 as indicated for an additional 12 h.Western blot was performed to determine the inhibitory effect of CAY10603 on NLRP3 inflammasome.3)HDAC6 knockout(KO)mice were generated and used to create STZ-induced model of DKD.The mice were divided into four groups:C57BL/6J wild type(WT)(n=6),HDAC6 KO(n=6),WT+STZ(n=10),and HDAC6 KO+STZ(n=9).Samples were collected 16 weeks after successful modeling and changes in uACR and renal pathology were evaluated accordingly.Results After 2 weeks of treatment,mice in the STZ+CAY10603 group exhibited reduction in uACR(P＜0.05)and inhibition of glomerular mesangium expansion(P＜0.05)compared with those of the mice in the STZ+Veh group.There was no statistically significant difference in the indicators between the CTL+Veh group and the CTL+CAY10603 group.In vitro cultured podocytes,compared with the control group,NLRP3 inflammasome activation was seen in the TGFβ group.CAY10603 treatment significantly inhibited the activation of NLRP3 in the dosage-dependent manner(P＜0.05).Compared with those of the WT group,the WT+STZ group showed increased uACR(P＜0.05),obvious glomerulosclerosis and loss of podocytes numbers.Compared with those of the WT+STZ group,the HDAC6 KO+STZ group showed effectively reduction of uACR(P＜0.05)and improvement in the renal pathological changes in mice.There was no significant difference in these aspects between the WT and HDAC6 KO groups.Conclusion Inhibition of HDAC6 alleviates proteinuria and podocyte injury in the mouse model of DKD by suppressing the activation of NLRP3 inflammasome.

ObjectiveTo observe the effect of twelve-week aerobic exercise on inhibitory control abilities and the change of brain activation in overweight children. MethodsFrom October to December, 2021, 20 overweight children from a primary school in Changping District were selected for a twelve-week aerobic exercise intervention. Their inhibitory control abilities were measured by Flanker task before and after intervention, while their brain activation levels during the task were detected by functional near infrared spectroscopy (fNIRS). ResultsThe interactions between task type and time of accuracy and reaction time in inconsistent tasks Flanker task were significant (F > 9.277, P < 0.05), with higher accuracy and lower reaction time of after intervention (P < 0.05). After intervention, ch1, ch2, ch3, ch6, and ch8 channels were activated by inconsistent tasks (P < 0.05). ConclusionA twelve-week aerobic exercise intervention could improve the inhibitory control ability, and increase the prefrontal cortex activation during inconsistent tasks in overweight children.

Objective To investigate the feasibility and clinical effectiveness of modified humeral supracondy-lar step－cut osteotomy(M－HSSCO)for cubitus varus in children.Methods Between June 2016 and June 2017,the study included 25 children (20 boys and 5 girls)presenting cubitus varus at the age of (8.5 ± 0.3)years old (6－11 years old)in the Children's Hospital of Nanjing Medical University.M－HSSCO was used in the osteotomy operation. The initial osteotomy was performed as a right triangle osteotomy.The inferior margin of this right triangle was outlined parallel to the joint line 0.5－1.0 cm above the olecranon fossa.Then,they were incised from the medial end of this parallel line,which made an angle between the first and second lines equal to the desired corrective humerus elbow－wrist angle.Next,from the lateral end of the second line,the third line perpendicular to the first line was cut.Finally, desired right triangle was outlined and removed.The second right triangle osteotomy from proximal line of osteotomy was similar as the above.The medial cortex and periosteum was used as a hinge and rotated to make two osteotomy parts combined steadily.The Bellemore evaluation system was used to evaluate the difference in carrying angle degree be-tween the affected side and normal side after osteotomy,and the different degree of flexion and extension in the elbow joint between preoperation and post－operation.Residual varus deformity and other complications were also evaluated. Results All patients were followed up for 6－18 months (mean 12 months).The preoperative ipsilateral carrying an-gle was－(15°－30°)[－(22.20 ± 1.40)°],the postoperative was 5°－10°[(7. 60 ± 0.47)°]and the final follow－up was 5°－9°[(7.30 ± 0.40)°].There were statistically significant differences in the flexion angles before surgery and after surgery (t＝23.34,P＝0.000).There were statistically significant differences in the flexion angles before surgery and at final follow－up (t＝23.14,P＝0.000).The mean range of joint motion of the elbow was －5°－130° [(132.5 ± 0.5)°]before surgery,0°－125°[(123.6 ± 1.4)°]after surgery and 0°－130°[(126.8 ± 1.3)°]at fi-nal follow－up.The carrying angle reduced did not happen by the final follow－up.There was no statistically significant difference between the flexion angles before surgery and after surgery or by final follow－up(P＞0. 05).Twenty－three (92.0%)patients got an excellent result,2 cases(18.0%)got a good result and no patient (0)got worse according to the criteria described by Bellemore.The first－class incision healing were achieved in all patients.There was no case with postoperative infection,instabilities,vascular and neural injury or any significant postoperative complications. Conclusions M－HSSCO is an effective osteotomy procedure which shows good feasibility and apparent therapeutic effect in treating cubitus varus in children.

Objective To evaluate and compare the imaging quality and diagnostic accuracy of two types of scanning techniques of 256-slice spiral CT angiography, prospective electrocardiogram(ECG)-gated sequence and non ECG-gated high-pitch sequence, used respectively for patients with Stanford type A aortic dissection (AAD) at the root of ascending aorta.Methods A retrospective study was conducted. Sixty-eight patients with AAD were definitely diagnosed by CT angiography were admitted to the Fifth Central Hospital of Tianjin from August 2015 to July 2017, and they were divided into two groups according to different scanning methods: 36 cases of AAD underwent prospective ECG-gated CT angiography (CTA) and 32 patients underwent non ECG-gated high-pitch CTA. A 3-grade scoring system was used to interpret the image quality of the aortic sinus, aortic valve junction zone and coronary artery opening. The CT value and noise value (SD) of aortic root were measured, the image signal to noise ratio (SNR) was calculated and compared with the discovery in exploratory operation; the patients' CTA imaging quality and the coincidence rate of lesion type diagnosis were compared between the two groups.Results All aortic CTA images could meet the diagnostic requirements. The imaging quality scores of aortic sinus, arotic valve junction zone and coronary artery opening images in ECG-gated CTA group were higher than those in non ECG-gated high-pitch CTA group (aortic sinus: 2.94±0.23 vs. 1.89±0.67, sinuscanal junction zone: 2.94±0.23 vs. 1.83±0.70, coronary artery opening images: 2.86±0.35 vs. 1.31±0.52, allP < 0.01); comparisons between the ECG-gated CTA group and non- ECG-gated CTA group in objective indexes, CT value, arotic SD value and SNR at the root of ascending aorta showed there were no statistically significant differences [the value of CT (HU): 425.20±94.38 vs. 439.29±86.78, the SD value of aorta (HU): 22.85±9.40 vs. 26.40±9.41, SNR: 21.23±8.16 vs. 19.70±9.98, allP > 0.05]. The coincidence rate between the diagnosis of AAD at the root of ascending aorta and the discovery in the exploratory operation in ECG-gated CTA group was higher than that in non ECG-gated CTA group [94.4% (34/36) vs. 68.8% (22/32),P < 0.01].Conclusion The diagnostic accuracy and image quality of AAD root of ascending aorta in prospective ECG-gated CTA group were significantly higher than those in non ECG-gated CTA group.