ObjectiveTo explore the effect of Shenge Bushen Capsules （SBC） on sexual development in normal 3-week-old mice. MethodsThe experiment consisted of two parts. In the first part， mice were divided into four groups： The control group and the low， medium， and high-dose SBC groups （234.7， 469.4， 938.7 mg·kg^-1， respectively）. In the second part， mice were divided into four groups： Control group， Pseudostellariae Radix polysaccharide （PRP） group， total flavonoids group， and SBC group， all receiving a dose of 469.4 mg·kg^-1. After 7 days of administration， the vaginal opening of female mice and the descent of testes and scrotum in male mice， as well as the ovarian and testicular organ indices， were observed. After 4 weeks of administration， female and male mice were housed together for 2 days， and the pregnancy rate of females was monitored. After delivery， the pregnant female mice continued receiving the treatment for 4 weeks， and the sexual development of their offspring， including vaginal opening， testicular descent， and organ indices of ovaries and testes， was observed. Serum sex hormones were measured by enzyme-linked immunosorbent assay （ELISA）， and the expression of gonadotropin-releasing hormone （GnRH） and growth hormone （GH） proteins in the hypothalamus was assessed by Western blot. ResultsCompared with the control group， there was no significant effect on the vaginal opening of female mice or the descent of testes in male mice after 7 days of SBC administration. After 4 weeks of administration， the pregnancy rate in the low-dose group was significantly reduced （P<0.05）， but no significant effects were observed in the other groups. The three doses of SBC did not significantly affect the ovarian or testicular organ indices， and there was no significant upregulation in the expression of GnRH or GH in the hypothalamus. The primary component of SBC， Pseudostellariae Radix polysaccharide， significantly reduced the vaginal opening in female mice after 7 days of administration （P<0.05）. After 4 weeks， the serum estradiol levels of non-pregnant female mice were decreased （P<0.05）， but there was no significant effect on the expression of GnRH or GH proteins in the hypothalamus of either male or female mice. Additionally， there were no significant effects on precocious puberty indicators， such as vaginal opening and testicular descent， in the offspring mice. ConclusionSBC does not significantly promote precocious puberty in young mice， and it does not have any noticeable effects on the pregnancy rate of adult mice or the sexual development of their offspring.

Metabolic-associated fatty liver disease (MAFLD) and osteoporosis (OP) are two very common metabolic diseases. A growing body of experimental evidence supports a pathophysiological link between MAFLD and OP. MAFLD is often associated with the development of OP. Rutaecarpine (RUT) is one of the main active components of Chinese medicine Euodiae Fructus. Our previous studies have demonstrated that RUT has lipid-lowering, anti-inflammatory and anti-atherosclerotic effects, and can improve the OP of rats. However, whether RUT can improve both fatty liver and OP symptoms of MAFLD mice at the same time remains to be investigated. In this study, we used C57BL/6 mice fed a high-fat diet (HFD) for 4 months to construct a MAFLD model, and gave the mice a low dose (5 mg·kg^-1) and a high dose (15 mg·kg^-1) of RUT by gavage for 4 weeks. The effects of RUT on liver steatosis and bone metabolism were then evaluated at the end of the experiment [this experiment was approved by the Experimental Animal Ethics Committee of Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences (approval number: IMB-20190124D₃03)]. The results showed that RUT treatment significantly reduced hepatic steatosis and lipid accumulation, and significantly reduced bone loss and promoted bone formation. In summary, this study shows that RUT has an effect of improving fatty liver and OP in MAFLD mice.

Aim To explore the effect of quercetin(Que)on ferroptosis and the potential mechanisms in an Erastin-induced ferroptosis model in chondrocytes.Methods A model of Erastin-induced ferroptosis was established in C28/I2 chondrocytes.Cells were treated with different concentrations of Que.Cell viability and cytotoxicity were assessed by MTT and LDH assays.The expression levels of Prdx6 and ferroptosis-related proteins ACSL4 and GPX4 in chondrocytes were deter-mined by Western blot.Lipid ROS production in chon-drocytes was measured by flow cytometry,while the changes in mitochondrial membrane potential were de-tected by RH123 staining.Prdx6 mRNA expression in chondrocytes was quantified by RT-qPCR.The chan-ges in the expression of the ferroptosis-related proteins ACSL4 and GPX4 were detected by immunofluores-cence staining.Results Compared to the Erastin-in-duced ferroptosis model group,Que significantly im-proved the viability of C28/I2 chondrocytes and re-duced cell cytotoxicity.It decreased the expression of the ferroptosis-related protein ACSL4 and increased the expression of GPX4.Que also inhibited the production of lipid ROS in chondrocytes and strengthened their mitochondrial membrane potential.In addition,the ex-pression of Prdx6 was significantly reduced in the Eras-tin group compared to the control group,while Que treatment upregulated the expression of Prdx6.Mean-while,the inhibitory effect of Que on chondrocyte fer-roptosis was reduced by the use of MJ33,an inhibitor of Prdx6.Conclusion Que can inhibit Erastin-induced ferroptosis of C28/I2 chondrocytes,possibly by upregu-lating Prdx6,and thus play a protective role in chon-drocytes.

Aim To investigate the promotional effects of neutrophil extracellular traps(NETs)on renal tissue damage and intestinal flora disruption induced by dia-betic kidney disease(DKD)and the potential mecha-nisms.Methods C57BL/6 mice were divided into:control group(NC),DNase Ⅰ control group(DNase Ⅰ)diabetic nephropathy group(DKD),and DNase Ⅰ treated group(DKD+DNase Ⅰ).The pathological changes of mouse kidney were observed by PAS,MAS-SON,and HE staining.The expression and distribu-tion of the relevant proteins of NETs in renal tissue of the mice in each group were observed by immunohisto-chemistry.The expression and distribution of coke-death-related proteins in the kidney tissues of mice in each group were observed by immunohistochemistry.The protein expression of NETs-related indexes,focal death-related indexes and NF-κB signaling pathway-re-lated indexes in kidney tissue of mice in each group were detected by immunoblotting.Results The ex-pression of indicators related to NETs was elevated in the DKD group,and their expression decreased after degradation of NETs by DNase Ⅰ(P＜0.01).Patho-logical staining results showed that the kidneys of DKD mice were structurally abnormal,and the structure was improved after degradation of NETs by DNase Ⅰ.The results of immunohistochemical staining and immunob-lotting showed that the expression of pyroptosis-related proteins in kidney tissues of mice in the DKD group was elevated compared with that in the control group(P＜0.01).NF-κB-related signaling pathway protein expression profile expression rose,and its expression decreased after degradation of NETs by DNase Ⅰ(P＜0.01)Conclusions NETs are generated in diabetic nephropathy and promote the onset of renal focal death and activation of the NF-κB signaling pathway,thereby exacerbating diabetes-induced kidney injury.

Objective To compare the efficacy of renal proximal renal artery denervation(pRDN)and full-length renal artery denervation(fRDN)for treatment of hypertension.Methods Fifty-six hypertensive patients were enrolled and randomly assigned to full-length renal artery denervation group(n=25)and proximal renal artery denervation group(n=31).After the procedure,24-hour ambulatory blood pressure monitoring(24 h-ABPM)at 6 months and office blood pressure at 12 months was recorded for statistical analysis.Results The blood pressure at follow-up reduced significantly in both groups,while there was no significant difference between groups.The baseline office blood pressure in fRDN group and pRDN group was(180±15)/(104±10)mmHg and(180±12)/(103±8)mmHg,respectively,which decreased to(142±9)/(82±7)mmHg and(143±10)/(83±6)mmHg at 12 months postoperatively(P＜0.001 within groups and P＞0.05 between groups).The baseline 24 h-ABPM in the two groups was(162±13)/(95±8)mmHg and(160±12)/(94±8)mmHg,respectively,which decreased to(142±11)/(83±7)mmHg and(141±8)/(81±7)mmHg at 6 months postoperatively(P＜0.001 within groups and P＞0.05 between groups).However,there was no significant difference in the reduction of office blood pressure and ambulatory blood pressure between the two groups.No treatment-related adverse events were observed.Conclusions pRDN has similar antihypertensive effect to fRDN.

Pancreatic polypeptide(PP),a pancreatic hormone containing 36 amino acids,plays impor-tant roles in the diagnosis and evaluation of pancreatic function,injury and diseases.In this study,a phage nanobody library against PP was constructed to screen specific PP nanobodies,which would be used to evaluate whether they have binding activity with PP antigen.After PP antigen with high purity was prepared by prokaryotic expression system,it was used to immunize alpaca to construct the nanobody li-brary against PP with high storage capacity and high abundance,from which 8 strains of PP nanobodies were obtained by phage display.One of nanobody strain(PP-VHH)was selected to be expressed in a prokaryotic expression system,which was induced overnight by IPTG.After purification and identifica-tion,the antigen-antibody binding activity and PP level in serum were detected by indirect ELISA and Sandwich ELISA methods,respectively.The results showed that PP-VHH had binding activity with PP,which could be used to detect PP in chicken and human serum.The Sandwich ELISA methods with R2 of the fitting curve 0.9868 could be used to detect PP concentrations of 48-55 pg/mL in the serum of chick-ens,while the concentrations of PP in human serum varied significantly.In summary,PP-VHH screened from nanobody library against PP could detect PP in serum,which would supply the basis for evaluation of abnormal pancreatic function and diagnosis of relative disease.

Objective:To explore the changes in number and immune function of mucosal-associated invariant T (MAIT) cells in peripheral blood of patients with newly diagnosed acute myeloid leukemia (AML),and its correlation with the occurrence and development of AML. Methods:Seventy-five clinical samples of patients with newly diagnosed AML and 48 healthy control samples in our hospital from January 2022 to February 2023 were included. Multiparametric flow cytometry was used to detect the number of MAIT cells,membrane surface markers,effector phenotypes and functional indicators in the samples. Results:Compared with healthy controls,the percentage of MAIT cells in CD3+T cells in peripheral blood of newly diagnosed AML patients was significantly reduced (P<0.001). The percentage of MAIT cells in all CD3+T cells in bone marrow of AML patients was similar to that in peripheral blood (P>0.05). Most of MAIT cells in peripheral blood of AML patients were effector memory T cells. Compared with healthy controls,the proportion of effector memory MAIT cells decreased (P<0.05),while the proportion of terminally differentiated effector memory MAIT cells and PD-1+MAIT cells increased significantly (both P<0.05). AML patients' peripheral blood MAIT cells expressed significantly higher levels of granzyme B and perforin than healthy controls (both P<0.05),and secreted significantly lower levels of cytokines such as gamma interferon and tumor necrosis factor α than healthy controls (both P<0.001). Conclusion:Compared with healthy controls,the proportion of MAIT cells in AML patients is reduced and the expression of functional markers is abnormal,suggesting that their function is impaired and may be involved in the occurrence and development of AML.

Objective:To investigates the role and mechanism of ferroptosis in combined burn-blast injury with acute lung injury in rats.Methods:This study was an experimental study. Twenty-four 8-week-old male Sprague-Dawley rats were divided into control group and experimental group by random number table method, each containing 12 animals. The rats in experimental group were anesthetized and subjected to explosion treatment to create the model of combined burn-blast injury with acute lung injury, whereas the rats in control group underwent sham injury. At 24 hours post injury, the pathological morphology of lung tissue was observed by hematoxylin-eosin staining and immunohistochemical staining. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 in the supernatant of bronchoalveolar lavage fluid (BALF) were detected by enzyme-linked immunosorbent assay. The arterial partial pressure of oxygen (PaO 2) and arterial partial pressure of carbon dioxide (PaCO 2) of abdominal aortic blood were measured by automatic animal blood gas analyzer. The lung tissue was weighed and the wet-dry weight ratio was calculated. The total protein concentration in BALF was measured by bicinchoninic acid assay. Lung injury was scored based on hematoxylin-eosin staining. The levels of oxidative stress factors, such as reactive oxygen species, malondialdehyde, superoxide dismutase (SOD), glutathione, and ferrous ion in lung tissue homogenate of rats were detected by related kits. The expression levels of ferroptosis-related molecule glutathione peroxidase 4 (GPX4), lipid peroxidation-related molecule 4-hydroxynonenal (4-HNE), and oxidative DNA damage-related molecule 8-hydroxydeoxyguanosine (8-OHdG) in lung tissue were detected by immunofluorescence and immunohistochemistry methods. Mitochondrial morphology in lung tissue cells was observed under transmission electron microscopy. The sample number was all 6. Results:At 24 hours post injury, the lung tissue structure of rats in control group was clear and complete, and the alveolar wall was normal; in experimental group, the lung tissue edema of rats was obvious, the alveolar wall became thicker, and the structure was not clear. At 24 hours post injury, compared with those in control group, the levels of TNF-α, IL-1β, and IL-6 in BALF supernatant of rats in experimental group were significantly increased (with t values of 3.96, 9.84, and 10.60, respectively, P<0.05); the wet-dry weight ratio of lung tissue, lung injury score, and total protein concentration in BALF of rats in experimental group were significantly increased (with t values of 6.91, 6.64, and 10.04, respectively, P<0.05), PaO 2 of abdominal aortic blood decreased significantly ( t=8.85, P<0.05) while PaCO 2 did not change significantly ( P>0.05); the levels of SOD and glutathione in the lung tissue homogenate of rats in experimental group were significantly decreased (with t values of 4.36 and 8.56, respectively, P<0.05), while the levels of reactive oxygen species, malondialdehyde, and ferrous ion were significantly increased (with t values of 11.55, 9.78, and 14.77, respectively, P<0.05). At 24 hours post injury, immunofluorescence staining and immunohistochemical staining showed that the expression levels of GPX4 in lung tissue of rats in experimental group were 0.245±0.024 and 0.786±0.240, respectively, which were significantly lower than 1.000±0.305 and 1.000±0.200 in control group (with t values of 6.05 and 2.60, respectively, P<0.05); the expression levels of 4-HNE in lung tissue of rats in experimental group were 5.93±1.05 and 2.21±0.23, respectively, which were significantly higher than 1.00±0.29 and 1.00±0.23 in control group (with t values of 11.13 and 9.16, respectively, P<0.05); the expression levels of 8-OHdG in lung tissue of rats in experimental group were 2.08±0.40 and 1.61±0.29, respectively, which were significantly higher than 1.00±0.40 and 1.00±0.26 in control group (with t values of 4.72 and 3.87, respectively, P<0.05). At 24 hours post injury, compared with that in control group, the density of mitochondrial double-layer membrane in the lung tissue cells of rats in experimental group increased, the outer membrane ruptured, and the crista decreased. Conclusions:In rats with combined burn-blast injury with acute lung injury, there is oxidative DNA damage in lung tissue cells, the imbalance of antioxidant system in lung tissue, and a decrease in the expression of GPX4, the key molecule against ferroptosis, suggesting that ferroptosis is involved in the pathophysiological process of this disease.

Objective To study the inner ear appearance on three-dimensional real inversion recovery(3D re-al IR)imaging in patients with sudden sensorineural hearing loss,and to investigate the relationship between blood-labyrinth barrier permeability and pathogenesis and prognosis of sudden hearing loss.Methods A total of 41 pa-tients with sudden sensorineural hearing loss received 3D real IR at 3.0 T MRI,and the signal intensity of inner ear were recorded.We respectively measured cochlear signal intensity in affected and healthy ears,and medullary signal intensity,and calculated the cochlear/medulla ratio(CM ratio)separately.On the basis of CM ratio,we evaluated hearing levels at initial and after treatment,and the relationship between CM ratio and hearing prognosis.Results Among the 41 patients,33 cases(occupying 80.48％)individually had a higher CM ratio in the affected ear than in the healthy one.The CM ratio of the affected side was not higher than that of the healthy side in 8 cases,and the ef-fective rate was 100％,eighteen cases had below 1.5 times the CM ratio in affected ears as in healthy ears,with 77.78％effective rate of treatment;seven cases had 1.5 to 1.75 times,with 100％effective rate;two cases had 1.75 to 2 times,with 50％effective rate;and the rest 14 cases had over 2 times,with 14.28％effective rate(P＜0.05).Conclusion The variation of blood-labyrinth barrier permeability in patients with sudden hearing loss can be read on the 3D Real IR,and it indicates that 80.48％patients have higher-intensity signals in lateral cochlea.The CM ratio can be adopted to evaluate the extent of inner ear damage of patients more accurately.As the CM ratio ri-ses,patients'prognosis become worse,and patients with over 1.75 times the CM ratio in the affected ear as in the healthy one,mostly suffer poor prognosis.

Streptococcus anginosus group(SAG)used to be considered as normal oral flora.With the advance-ment of diagnostic techniques,an increasing number of case reports confirm that SAG group can be an opportunistic pathogen causing a series of fatal infection.Therefore,this article reviews the diseases caused by SAG as opportu-nistic pathogens and its related pathogenic mechanisms.