ObjectiveTo investigate the mechanisms through which Shugan Bushen Yulin decoction ameliorates spermatogenic dysfunction and sperm quality degradation caused by propylthiouracil （PTU） via the endoplasmic reticulum stress pathway in rats. MethodsSixty male rats were randomly assigned into six groups： a control group， a model group， low- （6.75 g·kg^-1）， medium- （13.5 g·kg^-1）， and high-dose （27 g·kg^-1） Shugan Bushen Yulin decoction groups， and an L-carnitine （0.27 g·kg^-1） group， with ten rats in each group. Other groups except the control group were administrated with PTU （10 mg·kg^-1） by gavage for 12 consecutive days. After the completion of modeling， rats were administrated with corresponding agents or normal saline （control group） via gavage for 28 consecutive days. Twenty-four hours after the last administration， rats were sacrificed， and the body and organ （testis， epididymis， and seminal vesicle） weights were measured to calculate the organ indices. Hematoxylin-eosin staining was employed to observe the pathological changes in the testes and epididymis， and the testicular spermatogenic function of rats was scored. Enzyme-linked immunosorbent assay was employed to measure the levels of thyroid-stimulating hormone （TSH）， triiodothyronine （T3）， thyroxine （T4）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， testosterone （T）， estradiol （E₂）， and malondialdehyde （MDA） and the activities of superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） in the rat serum. Terminal deoxynucleotidyl transferase dUTP nick end labeling was adopted to assess the rate of testicular cell apoptosis. Western blot was conducted to determine the expression levels of glucose-regulated protein 78 （GRP78）， CCAAT/enhancer-binding protein homologous protein （CHOP）， phosphorylated protein kinase RNA-like endoplasmic reticulum kinase （p-PERK）， protein kinase RNA-like endoplasmic reticulum kinase （PERK）， phosphorylated eukaryotic translation initiation factor 2α kinase （p-EIF2α）， eukaryotic translation initiation factor 2α kinase （EIF2α）， and activating transcription factor 4 （ATF4） in the testicular tissue of rats. ResultsCompared with the control group， the model group exhibited reductions in volumes and indexes of testes， epididymides， and seminal vesicles （P<0.01）， pathological damage of testes and epididymides， and declines in spermatogenic function and sperm density and motility （P<0.01）. In addition， the model group demonstrated elevated serum levels of TSH and MDA， lowered levels of T3， T4， FSH， LH， T， and E₂， decreased activities of SOD and GSH-Px （P<0.05， P<0.01）， an increased apoptosis rate of testicular cells （P<0.01）， and up-regulated expression of GRP78， CHOP， p-PERK/PERK， p-EIF2α/EIF2α， and ATF4 in the testicular tissue （P<0.01）. Compared with the model group， Shugan Bushen Yulin decoction at different doses improved the morphology and indexes of testes， epididymides， and seminal vesicles （P<0.05）， restored the spermatogenic function （P<0.05， P<0.01）， increased the sperm density and motility （P<0.05， P<0.01）， and decreased the percentage of apoptotic testicular cells （P<0.05）. Furthermore， the decoction lowered the serum level of MDA， elevated the serum levels of FSH， LH， T， and E₂， and increased the activities of SOD and GSH-Px （P<0.05， P<0.01）. Moreover， the expression levels of GRP78， CHOP， p-PERK/PERK， p-EIF2α/EIF2α， and ATF4 in the testicular tissue were down-regulated （P<0.05， P<0.01）. ConclusionShugan Bushen Yulin decoction can ameliorate the spermatogenic dysfunction and sperm quality degradation induced by PTU in a rat model of hypothyroidism by repairing the testicular oxidative damage and regulating the PERK/EIF2α/ATF4 signaling pathway， thereby alleviating spermatogenic cell apoptosis.

Objective To establish the characteristic atlas of Shexiang Jiegu Capsule and determine the contents of seven active components （hydroxysafflor Yellow A, paeoniflorin, ferulic acid, naringin, ligustilide, catechin, epicatechin）. Methods Octadecyl silane bonded silica gel was used as the filling agent, the mobile phase was composed of methanol-0.05% phosphoric acid by gradient elution, the detection wavelength was 245 nm, flow rate was 1.0 ml/min, column temperature was 30℃. The similarity of the fifteen batches of sample was evaluated in line with the TCM Chromatographic Fingerprint （2012 edition）, and the contents of seven active components were determined. Results The HPLC fingerprint of Shexiang Jiegu Capsules was established. The similarity of fingerprint between fifteen batches of samples and control fingerprint was between 0.893 and 0.992. The results of methodological investigation for the determination of seven active components in fifteen batches of samples all met the requirements. Conclusion The established characteristic atlas of Shexiang Jiegu Capsules had high specificity and good repeatability, which could provide scientific basis for quality control of Shexiang Jiegu Capsules.

Objective To analyze the role and mechanism of PD-L1 in oral cancer metastasis based on TCGA and GEO databases.Methods The expression characteristics and clinical significance of the PD-L1 in oral cancer were analyzed using the TCGA database.PD-L1 mRNA levels were detected by quantitative reverse transcription-polymerase chain reaction(RT-qPCR)in various oral cancer cell lines.CCK-8,scrath test,Transwell-migration,and matrigel-invasion assays were employed to assess the effects of PD-L1 on proliferation,migration,and invasion of oral cancer cells.The interaction network between PD-L1 and functional genes in patients with oral cancer was constructed using STRING software and the GEO database,and key pathways were screened by Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.The regulatory relationship between PD-L1 and key genes was validated by RT-qPCR.Results TCGA data revealed that PD-L1 was highly expressed in patients with oral cancer and was correlated with lymph node metastasis(P＜0.01).PD-L1 was also highly expressed in oral cancer cell lines and its inhibition significantly inhibited the proliferation,migration,and invasion of Cal27 and SCC25 cells(P＜0.05).KEGG analysis indicated that PD-L1 activated the Janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway by upregulating C-X-C motif chemokine ligand(CXCL)9 and CXCL10,thereby promoting STAT1 expression to regulate oral cancer metastasis.Inhibition of the JAK/STAT pathway further suppressed the proliferation,migration,invasion,and expression of STAT1,CXCL9,and CXCL10 in Cal27 and SCC25 cells(P＜0.05).Conclusions PD-L1 may promote oral cancer cell proliferation,migration,and invasion by upregulating CXCL9 and CXCL10 to regulate the JAK/STAT pathway and enhance STAT1 expression,ultimately driving oral cancer growth and metastasis.

ObjectiveTo investigate the effects of Zuoguiwan on osteoclast activation induced by breast cancer and its mechanism. MethodsTo simulate breast cancer-induced osteoclastic bone metastasis， RAW264.7 cells were cultured in conditioned medium containing 50% supernatant of MDA-MB-231 breast cancer cells. The dosages of Zuoguiwan used in the experiment were sera containing 5% and 10% Zuoguiwan. Tartrate-resistant acid phosphatase （TRAP） staining was used to detect osteoclast activation. Enzyme-linked immunosorbent assay （ELISA） was used to measure Cathepsin K secretion from RAW264.7 cells. Real-time quantitative polymerase chain reaction （PCR） was used to detect the mRNA expression levels of osteocalcin （OCN） and bone sialoprotein （BSP）. Immunoprecipitation was employed to detect the interaction between Runt-related transcription factor 2 （Runx2） and core binding factor β subunit （CBF-β）. Western blot was used to assess the protein expression of Runx2， phosphorylated Runx2 （p-Runx2）， extracellular signal-regulated kinases 1/2 （ERK1/2）， p-ERK1/2， p38 mitogen-activated protein kinase （MAPK）， p-p38 MAPK， and CBF-β. ResultsCompared with the blank group， the MDA-MB-231 cell supernatant group showed a significant increase in TRAP-positive cell counts and Cathepsin K secretion. Meanwhile， the expression levels of p-Runx2， Runx2-CBF-β interaction， BSP and OCN mRNA， p-p38 MAPK， and p-ERK1/2 proteins were significantly decreased （P<0.01）. Compared with the MDA-MB-231 cell supernatant group， Zuoguiwan-containing sera significantly reduced TRAP-positive cell counts and Cathepsin K secretion （P<0.01）， significantly increased p-Runx2， BSP and OCN mRNA expression， as well as p-p38 MAPK and p-ERK1/2 protein levels， and promoted the interaction between Runx2 and CBF-β （P<0.01）. No significant change in Runx2 expression was observed. Compared to the blank group， the BVD-523 group showed significantly lower expression of p-p38 MAPK and p-ERK1/2 proteins （P<0.01）. Compared with the BVD-523 group， both low and high concentration Zuoguiwan-containing sera groups showed significantly higher p-p38 MAPK expression （P<0.01）， and the high concentration Zuoguiwan group also exhibited a significant increase in p-ERK1/2 expression （P<0.01）， while no statistical difference was found in the low-dose group. ConclusionZuoguiwan inhibits osteoclast activation by inducing phosphorylation of the key transcriptional regulator Runx2 in intra-osteoclast bone formation， and this process is closely associated with the activation of the p38 MAPK/ERK signaling pathway.

To review the clinical characteristics, short-term outcomes, and risk factors of patients with infective endocarditis(IE) who underwent surgical treatment at a single center, and to summarize treatment experience.

Objective To analyze the role and mechanism of PD-L1 in oral cancer metastasis based on TCGA and GEO databases.Methods The expression characteristics and clinical significance of the PD-L1 in oral cancer were analyzed using the TCGA database.PD-L1 mRNA levels were detected by quantitative reverse transcription-polymerase chain reaction(RT-qPCR)in various oral cancer cell lines.CCK-8,scrath test,Transwell-migration,and matrigel-invasion assays were employed to assess the effects of PD-L1 on proliferation,migration,and invasion of oral cancer cells.The interaction network between PD-L1 and functional genes in patients with oral cancer was constructed using STRING software and the GEO database,and key pathways were screened by Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.The regulatory relationship between PD-L1 and key genes was validated by RT-qPCR.Results TCGA data revealed that PD-L1 was highly expressed in patients with oral cancer and was correlated with lymph node metastasis(P＜0.01).PD-L1 was also highly expressed in oral cancer cell lines and its inhibition significantly inhibited the proliferation,migration,and invasion of Cal27 and SCC25 cells(P＜0.05).KEGG analysis indicated that PD-L1 activated the Janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway by upregulating C-X-C motif chemokine ligand(CXCL)9 and CXCL10,thereby promoting STAT1 expression to regulate oral cancer metastasis.Inhibition of the JAK/STAT pathway further suppressed the proliferation,migration,invasion,and expression of STAT1,CXCL9,and CXCL10 in Cal27 and SCC25 cells(P＜0.05).Conclusions PD-L1 may promote oral cancer cell proliferation,migration,and invasion by upregulating CXCL9 and CXCL10 to regulate the JAK/STAT pathway and enhance STAT1 expression,ultimately driving oral cancer growth and metastasis.

Objective:To establish a general method for the simultaneous determination of hydroxyphenyl esters and quaternary ammonium bacteriostatic agents in eye drops.Methods:The chromatographic analysis was per-formed on an Agilent C18 column(4.6 mm ×250 mm,5 μm)with 1％triethylamine solution(pH adjusted to 5.0 with phosphoric acid)as mobile phase A and methanol as mobile phase B.Gradient elution was performed at col-umn temperature of 40 ℃.The detection wavelength was 214 nm,the flow rate was 1 mL·min-1,and the injec-tion volume was 20 μL.Results:Methylparaben,ethylparaben,propylparaben,butylparaben,benzalkonium chlo-ride and benzalkonium bromide were 0.11-559.0,0.10-513.0,0.10-258.8,0.11-270.5,1.07-537.0 and 1.03-512.8 μg·mL-1,respectively.The linear range was good(r＞0.999).The average recoveries of meth-ylparaben,benzalkonium bromide and benzalkonium chloride were 104.7％(RSD=1.3％),102.6％(RSD=1.1％)and 100.9％(RSD=1.1％),respectively.The contents of bacteriostatic agent in 100 batches of eye drops from 36 varieties of 12 enterprises were determined,and the accurate results were obtained.Conclusion:This meth-od provides a reference for the content quality control and safety evaluation of bacteriostatic agents in eye drops.

Objective To systematically review the incidence and influencing factors of refeeding syndrome(RFS)in critically ill patients,and provide references for early identification of RFS and formulation of preventive measures.Methods Computerized searches were conducted for studies on RFS in critically ill patients in the databases of China National Knowledge Infrastructure(CNKI),Wanfang,VIP,CBM,PubMed,Embase,Web of Science,CINAHL,Cochrane Library from inception to May 29th,2024.Data analysis was performed using Stata 16.0 software.Results A total of 29 articles with 5 720 participants were included.The Meta-analysis showed that the incidence of RFS in critically ill patients was 33.68％.The subgroup analysis showed that the incidence of RFS in critically ill patients was higher in studies conducted in 2020 or later(38.22％),in the Americas(36.39％),and with only electrolyte changes as the diagnostic basis(37.51％).Risk factors for RFS in critically ill patients included higher acute physiological and chronic health evaluation Ⅱ scores(OR=1.41),higher sequential organ failure assessment scores(OR=1.29),initiation of feeding within 48 h of ICU admission(OR=3.36),age ≥60 years(OR＝2.82),diabetes mellitus(OR=3.53),pre-albumin concentration＜150 g/L(OR=5.53),albumin concentration＜30 g/L(OR=3.26),caloric intake＞25％standard calories(OR=2.86),enteral solution temperature of 36～38 ℃(OR=2.32),feeding rate＞50 ml/h(OR=3.76),fasting time ≥2 d before feeding(OR=2.46),history of alcoholism(OR=2.64).Conclusion The incidence of RFS in critically ill patients is high and there are many influencing factors.Nurses should improve their awareness and attention to RFS,accurately identify high-risk groups and risk factors,and adopt a multidisciplinary collaborative model to develop whole-course,detailed and personalized intervention measures to prevent RFS.

Objective We aimed to investigate the effect of ferroptosis on Shenling Baizhu Powder,a compound prescription of Chinese herbal medicine,in improving testicular spermatogenic function in hyperuricemic mice with spermatogenic dysfunction. Methods Sixty BALB/c mice were randomly divided into normal group,model group,Shenling Baizhu Powder high-,medium-,and low-dose groups (20.14,10.07,5.04 g/kg,by gavage),and ferrostatin-1(Fer-1) group (0.8 mg/kg,by tail vein injection),with 10 mice each group. Except for the normal group,the other groups were intraperitoneally injected with potassium oxonate suspension[600mg/(kg·d)]for 7 days to establish the hyperuricemic model,and then the corresponding intervention was given for consecutive 14 days. Content of serum uric acid (UA),testicular Fe2+,reduced glutathione (GSH),malondialdehyde (MDA) and superoxide dismutase (SOD) activity were detected by biochemical method. Epididymal and testicular indices were measured. The spermatogenic function of testes was evaluated by eosin-hematoxylin staining. Sperm quality was detected by an automatic animal sperm analyzer. Prussian blue staining was used to detect iron deposition in testicular tissue. Immunohistochemistry was used to detect the related protein expressions of acyl-coenzyme A synthetase long-chain family member 4 (ACSL4) and glutathione peroxidase 4 (GPX4) in testicular tissue. Western blotting was used to detect the related protein expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1(HO-1)/GPX4 signaling pathway in testicular tissue. Results Compared with the normal group,the contents of serum UA,MDA,and Fe2+in the testis tissue of the model group were increased,the GSH content and SOD activity were decreased,the epididymal and testicular index,testicular spermatogenic function,sperm density and activity rate were decreased,and the iron deposition and ACSL4 protein expression in the testis tissue were increased. The expressions of kelch-like ECH-associated protein-1 (Keap1) and Nrf2 were increased. The expressions of nuclear Nrf2,HO-1,GPX4,and recombinant solute carrier family 7 member 11 (SLC7A11) protein were decreased (P<0.01). Compared with the model group,the above indexes in the Shenling Baizhu Powder groups and the Fer-1 group were improved to varying degrees (P<0.05,P<0.01). Conclusion Shenling Baizhu Powder can inhibit the ferroptosis of testicular cells through the Nrf2/HO-1/GPX4 signaling pathway,and improve the testicular spermatogenic function of mice with hyperuricemia spermatogenic dysfunction.

ObjectiveTo observe the clinical efficacy and safety of Shugan Bushen Yulin Decoction （疏肝补肾毓麟汤， SBYD） in the treatment of asthenospermia and infertility with liver constraint and kidney deficiency. MethodsA multicenter， randomized， controlled clinical study was conducted in three hospitals in central China. Totally 95 patients with asthenospermia and infertility were controlled. According to random number table， the patients were divided into treatment group （47 cases） and control group （48 cases）. The control group was given vitamin E soft capsules （100 mg per time， twice daily） orally， and the treatment group was given SBYD （one dose daily， 30 min after breakfast and dinner， about 200 ml each time） orally. The course of treatment was 12 weeks in both groups. After the treatment， the sperm concentration， percentage of forward motile sperm （PR）， and percentage of total sperm activity， that is PR + percentage of non-progressively motile sperm （NP） were compared between groups， and the clinical efficacy was judged. Traditional Chinese medicine （TCM） syndrome score （inlcuding 6 single symptom scores and total symptoms score） and 21-item depression， anxiety， and stress scale （DASS-21） scores （including depression， anxiety and stress scores） were compared between the two groups before and after treatment. The patients were followed up for 6 months， and the pregnancy status of spouse between groups was compared. The occurrence of adverse events and vital signs during the trial were recorded for safety assessment. ResultsTwo cases in the treatment group and three cases in the control group dropped out. Finally， 45 cases in each group were included in the statistical analysis. The total effective rate was 86.67% （39/45） in the treatment group， significantly higher than 73.74% （33/45） in the control group （P＜0.05）. After treatment， PR and PR + NP significantly increased in both groups， and were much higher in the treatment group than in the control group （P＜0.05）. The scores of lumbar and knee soreness， emotional disturbance， cold sperm， chest and rib-side and lesser abdomen distension and pain， frequent sighing， and the total TCM syndrome score in the treatment group decreased after treatment （P＜0.05）； and except for cold sperm， the scores of other symptoms mentioned above as well as the total TCM syndrome score in the treatment group were lower than those in the control group （P＜0.05）. The scores of depression， anxiety and stress of DASS-21 in the treatment group decreased after treatment， and were lower than those in the control group （P＜0.05）. After 6 months of follow-up， the spouses of 5 patients in the treatment group and 3 patients in the control group got pregnant （P＞0.05）. No adverse events occurred in both groups during the treatment， and their vital signs were stable. ConclusionSBYD can obviously improve sperm vitality and effectively improve the clinical symptoms， anxiety and depression， and is safe in treating asthenospermia and infertility with liver constraint and kidney deficiency.