Fifty whitespotted bamboo sharks (Chiloscyllium playgiosum) of both sexes were used to establish a large capacity variable domain of the new antigen receptor (VNAR) library with a total capacity of over 10⁹ colony-forming units (CFU). It was applied to screen VNARs against human serum albumin (HSA) and human transcription factor EB (TFEB), respectively. Meanwhile, VNAR libraries specific to HSA and TFEB with capacities above 10⁸ CFU were obtained following conventional immunization. These two approaches were systematically studied in terms of VNAR yield and composition. By comparing the VNAR sequences obtained from naïve and antigen-immunized libraries, we found that the complementary-determining region 3 (CDR3) of the former differs in composition from that of the latter. It shares a higher degree of homology with the naïve library. Meanwhile, the binding efficiency assessed by ELISA is also different between the naïve and antigen-immunized libraries. The binding of VNARs from the TFEB-immunized library appeared to surpass that observed with the naïve libraries, whereas the performance of VNARs from the HSA-immunized library indicated that both the immunized and naïve libraries for HSA had positive binding responses in polyclonal and monoclonal ELISA. The results are useful to develop novel diagnostic and therapeutic products based on shark VNARs.

Objective:To explore the role of the expression of peroxisome proliferator-activated receptor β/δ(PPARβ/δ)in the anti-inflammatory mechanism of acupuncture(AC)against cerebral ischemia-reperfusion injury(CIRI)in rat hippocampal tissue.Methods:A total of 60 SD rats were randomly divided into Sham group,CIRI group,CIRI+AC group and CIRI+AC+PPARβ/δsiRNA group.The CIRI rat model was prepared by the method of middle cerebral artery occlusion,and acupuncture treatment was used for intervention.The neurological function of rats was assessed by the modified neurological severity score(mNSS)method after 72 hours of intervention.The brain in-farction volume ratio was observed by 2,3,5-triphenyltetrazolium chloride(TTC)staining method.The pathological morphological changes of brain tissue were observed by hematoxylin-eosin(HE)staining.The expression levels of PPARβ/δ,IL-1β,and IL-10 mRNA in the hippocampal region were detected by RT-qPCR.Results:Compared with the Sham group,the mNSS scores and the brain infarction volume ratio in all modeling groups were significantly in-creased.Compared with the CIRI group,the mNSS scores and the brain infarction volume ratio in the CIRI+AC group were significantly reduced.The expression levels of PPARβ/δ and IL-10 in the hippocampus were significantly upregu-lated,while the expression level of IL-1β was significantly downregulated(P<0.01),with scattered inflammatory cell infiltration observed.Compared with the CIRI+AC group,there was no significant difference in the mNSS scores and the brain infarction volume ratio in the CIRI+AC+PPARβ/δsiRNA group.The expression level of PPARβ/δ in the hippocampus was significantly downregulated(P<0.01),while no significant difference was found in the expression level of IL-10 and IL-1β mRNA,with a small amount of inflammatory cell infiltration observed.Conclusion:Acupunc-ture ameliorates the recovery of neurological function and the ratio of cerebral infarction volume in rats with CIRI,and its mechanism may be associated with the upregulation of PPARβ/δ expression,thereby reducing inflammatory respon-ses and exerting neuroprotective effects.

Objective:To investigate the effects of acupuncture intervention on the expression of circRNA_017109 and miR-138-5p in the hippocampal region of rats with cerebral ischemia-reperfusion injury(CIRI),and to explore the potential mechanism of acupuncture in treating CIRI.Methods:Forty-five Sprague-Dawley(SD)rats were divided into sham group,middle cerebral artery occlusion/reperfusion(MCAO/R)group,and acupuncture(MCAO/R+AC)group.The MCAO/R and MCAO/R+AC groups were subjected to the modified Longa suture method to establish the MCAO/R model.The MCAO/R+AC group received acupuncture at the"Dazhui","Renzhong",and"Baihui"acu-points.Neurological impairment was assessed via the modified neurological severity score(mNSS).while cerebral infarct volume was assessed through TTC staining.Histological alterations in brain tissue were examined via HE stai-ning,and the expression of circRNA_017109 and miR-138-5p in the hippocampal area were determined using RT-qPCR.Results:The mNSS scores and cerebral infarct volume ratios in the model groups were markedly higher than those in the sham group(P<0.01),but these were significantly reduced in MCAO/R+AC group compared to the MCAO/R group(P<0.01).HE staining showed severe necrosis and inflammation in the MCAO/R group's hippocam-pus,while the MCAO/R+AC group had minimal necrosis and scattered inflammation.RT-qPCR results indicated that the expression of circRNA_017109 and miR-138-5p was downregulated in the MCAO/R group,while MCAO/R+AC group upregulated the expression of circRNA_017109 and miR-138-5p(P<0.05,P<0.01).Conclusion:Acupunc-ture can improve neurological deficits,reduce cerebral infarction volume,and alleviate inflammatory cell infiltration in CIRI rats by upregulating the expression of circRNA_017109 and miR-138-5p,thereby exerting neuroprotective effects.

Objective:To explore the role of the expression of peroxisome proliferator-activated receptor β/δ(PPARβ/δ)in the anti-inflammatory mechanism of acupuncture(AC)against cerebral ischemia-reperfusion injury(CIRI)in rat hippocampal tissue.Methods:A total of 60 SD rats were randomly divided into Sham group,CIRI group,CIRI+AC group and CIRI+AC+PPARβ/δsiRNA group.The CIRI rat model was prepared by the method of middle cerebral artery occlusion,and acupuncture treatment was used for intervention.The neurological function of rats was assessed by the modified neurological severity score(mNSS)method after 72 hours of intervention.The brain in-farction volume ratio was observed by 2,3,5-triphenyltetrazolium chloride(TTC)staining method.The pathological morphological changes of brain tissue were observed by hematoxylin-eosin(HE)staining.The expression levels of PPARβ/δ,IL-1β,and IL-10 mRNA in the hippocampal region were detected by RT-qPCR.Results:Compared with the Sham group,the mNSS scores and the brain infarction volume ratio in all modeling groups were significantly in-creased.Compared with the CIRI group,the mNSS scores and the brain infarction volume ratio in the CIRI+AC group were significantly reduced.The expression levels of PPARβ/δ and IL-10 in the hippocampus were significantly upregu-lated,while the expression level of IL-1β was significantly downregulated(P<0.01),with scattered inflammatory cell infiltration observed.Compared with the CIRI+AC group,there was no significant difference in the mNSS scores and the brain infarction volume ratio in the CIRI+AC+PPARβ/δsiRNA group.The expression level of PPARβ/δ in the hippocampus was significantly downregulated(P<0.01),while no significant difference was found in the expression level of IL-10 and IL-1β mRNA,with a small amount of inflammatory cell infiltration observed.Conclusion:Acupunc-ture ameliorates the recovery of neurological function and the ratio of cerebral infarction volume in rats with CIRI,and its mechanism may be associated with the upregulation of PPARβ/δ expression,thereby reducing inflammatory respon-ses and exerting neuroprotective effects.

Objective:To investigate the effects of acupuncture intervention on the expression of circRNA_017109 and miR-138-5p in the hippocampal region of rats with cerebral ischemia-reperfusion injury(CIRI),and to explore the potential mechanism of acupuncture in treating CIRI.Methods:Forty-five Sprague-Dawley(SD)rats were divided into sham group,middle cerebral artery occlusion/reperfusion(MCAO/R)group,and acupuncture(MCAO/R+AC)group.The MCAO/R and MCAO/R+AC groups were subjected to the modified Longa suture method to establish the MCAO/R model.The MCAO/R+AC group received acupuncture at the"Dazhui","Renzhong",and"Baihui"acu-points.Neurological impairment was assessed via the modified neurological severity score(mNSS).while cerebral infarct volume was assessed through TTC staining.Histological alterations in brain tissue were examined via HE stai-ning,and the expression of circRNA_017109 and miR-138-5p in the hippocampal area were determined using RT-qPCR.Results:The mNSS scores and cerebral infarct volume ratios in the model groups were markedly higher than those in the sham group(P<0.01),but these were significantly reduced in MCAO/R+AC group compared to the MCAO/R group(P<0.01).HE staining showed severe necrosis and inflammation in the MCAO/R group's hippocam-pus,while the MCAO/R+AC group had minimal necrosis and scattered inflammation.RT-qPCR results indicated that the expression of circRNA_017109 and miR-138-5p was downregulated in the MCAO/R group,while MCAO/R+AC group upregulated the expression of circRNA_017109 and miR-138-5p(P<0.05,P<0.01).Conclusion:Acupunc-ture can improve neurological deficits,reduce cerebral infarction volume,and alleviate inflammatory cell infiltration in CIRI rats by upregulating the expression of circRNA_017109 and miR-138-5p,thereby exerting neuroprotective effects.

Objective:To describe the distribution of lipoprotein (a) [Lp(a)] levels in non-arteriosclerotic cardiovascular disease (ASCVD) population in China and explore its influencing factors.Methods:This study was based on a nested case-control study in the CKB study measured plasma biomarkers. Lp(a) levels was measured using a polyclonal antibody-based turbidimetric assay certified by the reference laboratory and ≥75.0 nmol/L defined as high Lp(a). Multiple logistic regression model was used to examine the factors related to Lp(a) levels.Results:Among the 5 870 non-ASCVD population included in the analysis, Lp(a) levels showed a right-skewed distribution, with a M ( Q1, Q3) of 17.5 (8.8, 43.5) nmol/L. The multiple logistic regression analysis found that female was associated with high Lp(a) ( OR=1.23, 95% CI: 1.05-1.43). The risk of increased Lp(a) levels in subjects with abdominal obesity was significantly reduced ( OR=0.68, 95% CI: 0.52-0.89). As TC, LDL-C, apolipoprotein A1(Apo A1), and apolipoprotein B(Apo B) levels increased, the risk of high Lp(a) increased, with OR (95% CI) for each elevated group was 2.40 (1.76-3.24), 2.68 (1.36-4.93), 1.29 (1.03-1.61), and 1.65 (1.27-2.13), respectively. The risk of high Lp(a) was reduced in the HDL-C lowering group with an OR (95% CI) of 0.76 (0.61-0.94). In contrast, an increase in TG levels and the ratio of Apo A1/Apo B(Apo A1/B) was negatively correlated with the risk of high Lp(a), with OR (95% CI) of 0.73 (0.60-0.89) for elevated triglyceride group, and OR (95% CI) of 0.60 (0.50-0.72) for the Apo A1/B ratio increase group (linear trend test P≤0.001 except for Apo A1). However, no correlation was found between Lp(a) levels and lifestyle factors such as diet, smoking, and physical activity. Conclusions:Lp(a) levels were associated with sex and abdominal obesity, but less with lifestyle behaviors.

Osteoporosis (OP) is a systemic skeletal disease that primarily affects the elderly population, which greatly increases the risk of fractures. Here we report that Kindlin-2 expression in adipose tissue increases during aging and high-fat diet fed and is accompanied by decreased bone mass. Kindlin-2 specific deletion (K2KO) controlled by Adipoq-Cre mice or adipose tissue-targeting AAV (AAV-Rec2-CasRx-sgK2) significantly increases bone mass. Mechanistically, Kindlin-2 promotes peroxisome proliferator-activated receptor gamma (PPARγ) activation and downstream fatty acid binding protein 4 (FABP4) expression through stabilizing fatty acid synthase (FAS), and increased FABP4 inhibits insulin expression and decreases bone mass. Kindlin-2 inhibition results in accelerated FAS degradation, decreased PPARγ activation and FABP4 expression, and therefore increased insulin expression and bone mass. Interestingly, we find that FABP4 is increased while insulin is decreased in serum of OP patients. Increased FABP4 expression through PPARγ activation by rosiglitazone reverses the high bone mass phenotype of K2KO mice. Inhibition of FAS by C75 phenocopies the high bone mass phenotype of K2KO mice. Collectively, our study establishes a novel Kindlin-2/FAS/PPARγ/FABP4/insulin axis in adipose tissue modulating bone mass and strongly indicates that FAS and Kindlin-2 are new potential targets and C75 or AAV-Rec2-CasRx-sgK2 treatment are potential strategies for OP treatment.

Nighteen people at a restaurant experienced dizziness headaches and other discomforts in six days. According to the description method, the time and location distribution were found to be concentrated. A second Investigation was conducted at the same time as the onset of the case, the test found that the carbon monoxide concentration of second floor up to 539 mg/m 3. The on-site testing found that when 2 steam generator in snack room on the first floor turned on, the carbon monoxide concentration on the top of elevator on the second floor was 1225.0 mg/m 3. After the accident, the restaurant replaced a steam generator, the carbon monoxide concentration on the top of the new and old steam generator were 350 mg/m 3 and >1 000 mg/m 3, respectively. After the steam generators were fitted with exhaust smoke pipe and exhasust hood, the carbon monoxide concentrations of on the top of the vegetable transfer elevator and the room on the second floor were both 0.4 mg/m 3, and there were no cases of recurrence. It was determined that this was a carbon monoxide poisoning incident caused by a high concentration of carbon monoxide emitted by the steam generators, which spread to the second floor of the private room through the vegetable transfer elevator.

Nighteen people at a restaurant experienced dizziness headaches and other discomforts in six days. According to the description method, the time and location distribution were found to be concentrated. A second Investigation was conducted at the same time as the onset of the case, the test found that the carbon monoxide concentration of second floor up to 539 mg/m 3. The on-site testing found that when 2 steam generator in snack room on the first floor turned on, the carbon monoxide concentration on the top of elevator on the second floor was 1225.0 mg/m 3. After the accident, the restaurant replaced a steam generator, the carbon monoxide concentration on the top of the new and old steam generator were 350 mg/m 3 and >1 000 mg/m 3, respectively. After the steam generators were fitted with exhaust smoke pipe and exhasust hood, the carbon monoxide concentrations of on the top of the vegetable transfer elevator and the room on the second floor were both 0.4 mg/m 3, and there were no cases of recurrence. It was determined that this was a carbon monoxide poisoning incident caused by a high concentration of carbon monoxide emitted by the steam generators, which spread to the second floor of the private room through the vegetable transfer elevator.