Aim To explore the mechanism of Lycium barbarum glycopeptide(LbGP)improving aging in rat primary ovarian granulosa cells.Methods This study divided the cells into a normal group,a DOX group,and four different LbGP concentration treatment groups post-DOX intervention.Results Cell proliferation was assessed using CCK-8,EDU,and Ki67 assays,while aging markers and mitochondrial function-related fac-tors were detected using immunofluorescence and West-ern blotting.The results showed that,compared to the DOX group,LbGP treatment significantly increased cell viability(P＜0.05)and promoted proliferation(P＜0.05).Post LbGP treatment,the β-galactosidase-posi-tive area in cells was significantly reduced compared to the DOX group(P＜0.05).Immunofluorescence re-sults indicated that,compared to the DOX group,levels of p21 and γH2AX significantly decreased(P＜0.05),while pRB increased(P＜0.05)after LbGP treatment.Western blot results showed that,compared to the DOX group,the aging phenotype proteins p21 and p53 significantly decreased(P＜0.05),and pRB notably increased(P＜0.05)in the LbGP treatment group.The release of cytC into the cytoplasm and the activated caspase-9 significantly decreased(P＜0.05);levels of CAMKK2,pAMPK,and mitochondrial calcium homeostasis regulator MCU increased(P＜0.05);nuclear energy metabolism-related proteins SirT1,PGC1α/β and ATP5A1 significantly increased(P＜0.05);compared to the DOX group,ROS levels significantly decreased after LbGP treatment(P＜0.05).Conclusions The results suggest that LbGP can ameliorate DOX-induced aging in rat primary ovar-ian granulosa cells,potentially through the upregulation of the CAMKKβ/AMPK signaling pathway,thereby im-proving mitochondrial calcium homeostasis and increas-ing the expression levels of cell energy metabolism-re-lated regulatory proteins.This provides an experimen-tal basis for LbGP's potential role in supporting the im-provement of ovarian function.

Objective To observe the origin and course of the obturator artery(OA),so as to provide anatomical reference for reducing hemorrhage during pelvic surgery and pubic fracture fixation.Methods A total of 65 human hemi-pelvises specimens with intact structure were dissected to observe the origin,course and other variations of OA.Measure the length of the inner section of OA basin and the outer diameter at the origin,etc.Results OA originated from the internal iliac artery in 57 cases(87.7%),including 3 cases(4.6%)of the superior gluteal artery,5 cases(7.7%)of the inferior gluteal artery,3 cases(4.6%)of the external iliac artery and 5 cases(7.7%)of the inferior epigastric artery.OA participated in the formation of the arterial trunk in 3 cases(4.6%).The length of the pelvic segment of the OA in male and female was(50.87±15.41)mm and(51.71±14.19)mm,respectively,with no statistically significant difference between them(P>0.05).The outer diameters at the origin of the OA in male and female were(2.79±1.05)mm and(2.35±0.86)mm,and there was no statistically significant difference between them(P>0.05).Conclusion OA mainly originated from the anterior trunk of the internal iliac artery,with a few OA originated from the branches of the posterior trunk or the inferior epigastric artery,or participated in the formation of the arterial trunk.In pelvic surgery involving OA area,attention should be paid to the length of its pelvic segment and the outer diameter at the origin of OA,so as to better locate and protect blood vessels during surgery.

Objective To observe the origin and course of the obturator artery(OA),so as to provide anatomical reference for reducing hemorrhage during pelvic surgery and pubic fracture fixation.Methods A total of 65 human hemi-pelvises specimens with intact structure were dissected to observe the origin,course and other variations of OA.Measure the length of the inner section of OA basin and the outer diameter at the origin,etc.Results OA originated from the internal iliac artery in 57 cases(87.7%),including 3 cases(4.6%)of the superior gluteal artery,5 cases(7.7%)of the inferior gluteal artery,3 cases(4.6%)of the external iliac artery and 5 cases(7.7%)of the inferior epigastric artery.OA participated in the formation of the arterial trunk in 3 cases(4.6%).The length of the pelvic segment of the OA in male and female was(50.87±15.41)mm and(51.71±14.19)mm,respectively,with no statistically significant difference between them(P>0.05).The outer diameters at the origin of the OA in male and female were(2.79±1.05)mm and(2.35±0.86)mm,and there was no statistically significant difference between them(P>0.05).Conclusion OA mainly originated from the anterior trunk of the internal iliac artery,with a few OA originated from the branches of the posterior trunk or the inferior epigastric artery,or participated in the formation of the arterial trunk.In pelvic surgery involving OA area,attention should be paid to the length of its pelvic segment and the outer diameter at the origin of OA,so as to better locate and protect blood vessels during surgery.

Objective To investigate the size variation in the first metatarsal of extant primates.Methods In this study,we analyzed 135 first metatarsal specimens across 12 primate genera,quantifying eight linear measurements,articular surface areas,mid-shaft cross-sectional area,total surface area,volume,and derived indices.Multivariate patterns were assessed through mean-based correspondence analysis and principal component analysis(PCA).Results Eulemur,Otolemur,Cebus,and Perodicticus exhibited a relatively high metatarsal surface-area-to-volume ratio.Perodicticus additionally showed a low articular facet index.Propithecus,Colobus,and Macaca displayed lower values for metatarsal shaft robusticity,the ratio of shaft cross-sectional area to base articular surface area,and the proximal articular facet index.Nasalis possessed a relatively high articular facet index.Pongo,Pan,Gorilla,and Homo sapiens were characterized by higher metatarsal shaft robusticity and a lower metatarsal surface-area-to-volume ratio.Principal component analysis revealed that the 12 extant primate genera could be broadly divided into two groups.Group 1 comprised Pongo,Pan,Gorilla and H.sapiens,although H.sapiens formed a distinct cluster relative to the extant great apes.Group 2 included Eulemur,Otolemur,Perodicticus,Propithecus,Cebus,Colobus,Macaca and Nasalis.Conclusion The relative sizedistribution of the first metatarsal provides some reference value for classifying extant primates.However,it demonstrates no clear correlations with specific locomotor patterns or foot grasping ability.

Objective To observe the effects of wheat grain moxibustion on the hippocampal ERK/CREB-BDNF/TrkB signaling pathway and neurotransmitters in rats with perimenopausal depressive disorder(PDD);To explore its mechanism in treating PDD.Methods Totally 24 female SD rats were randomly divided into sham-operation group,model group,Western medicine group and wheat grain moxibustion group,with 6 rats in each group.The PDD rat model was established by bilateral ovariectomy and chronic unpredictable mild stress.The wheat grain moxibustion group received moxibustion at the"Zhongwan","Xiawan","Qihai","Guanyuan"acupoints,with a conical moxa cone for intervention,6 cones per acupoint;the Western medicine group was given fluoxetine hydrochloride solution 1.8 mg/kg by gavage,once a day,for consecutive 21 d.Before and after modeling and after intervention,the body mass,sucrose water preference rate and immobility time in the forced swimming test of the rats were recorded;Transmission electron microscopy was used to observe the synaptic structure in hippocampal tissue;ELISA was used to detect the expressions of 5-HT,GABA and Glu in hippocampal tissue;Western blot was used to detect the expressions of ERK,p-ERK,CREB,p-CREB,BDNF and TrkB proteins in hippocampal tissue.Results Compared with the sham-operation group,the body mass and sucrose water preference rate of the model group decreased,and the immobility time in the forced swimming test was prolonged(P<0.01);the number of synaptic vesicles in hippocampus decreased,the thickness of the postsynaptic density decreased,the aggregation of transduction-related proteins decreased,the synaptic cleft increased;the contents of 5-HT and GABA in hippocampal tissue decreased,the content of Glu increased,and the expressions of p-ERK,p-CREB,BDNF and TrkB proteins decreased(P<0.01).Compared with the model group,the body mass and sucrose water preference rate of the rats in Western medicine group and wheat grain moxibustion group increased,and the immobility time in the forced swimming test was shortened(P<0.01);the area of the postsynaptic density in the hippocampus increased,the synaptic cleft reduced,the contents of 5-HT and GABA in hippocampal tissue increased,the content of Glu decreased,and the expressions of p-ERK,p-CREB,BDNF and TrkB proteins increased(P<0.01).Conclusion Wheat grain moxibustion can effectively alleviate depressive-like behavior in PDD rats,and its mechanism may be related to regulating neurotransmitter expressions and mediating the ERK/CREB-BDNF/TrkB signaling pathway,thereby regulating hippocampal neuroplasticity.

Aim To explore the mechanism of Lycium barbarum glycopeptide(LbGP)improving aging in rat primary ovarian granulosa cells.Methods This study divided the cells into a normal group,a DOX group,and four different LbGP concentration treatment groups post-DOX intervention.Results Cell proliferation was assessed using CCK-8,EDU,and Ki67 assays,while aging markers and mitochondrial function-related fac-tors were detected using immunofluorescence and West-ern blotting.The results showed that,compared to the DOX group,LbGP treatment significantly increased cell viability(P＜0.05)and promoted proliferation(P＜0.05).Post LbGP treatment,the β-galactosidase-posi-tive area in cells was significantly reduced compared to the DOX group(P＜0.05).Immunofluorescence re-sults indicated that,compared to the DOX group,levels of p21 and γH2AX significantly decreased(P＜0.05),while pRB increased(P＜0.05)after LbGP treatment.Western blot results showed that,compared to the DOX group,the aging phenotype proteins p21 and p53 significantly decreased(P＜0.05),and pRB notably increased(P＜0.05)in the LbGP treatment group.The release of cytC into the cytoplasm and the activated caspase-9 significantly decreased(P＜0.05);levels of CAMKK2,pAMPK,and mitochondrial calcium homeostasis regulator MCU increased(P＜0.05);nuclear energy metabolism-related proteins SirT1,PGC1α/β and ATP5A1 significantly increased(P＜0.05);compared to the DOX group,ROS levels significantly decreased after LbGP treatment(P＜0.05).Conclusions The results suggest that LbGP can ameliorate DOX-induced aging in rat primary ovar-ian granulosa cells,potentially through the upregulation of the CAMKKβ/AMPK signaling pathway,thereby im-proving mitochondrial calcium homeostasis and increas-ing the expression levels of cell energy metabolism-re-lated regulatory proteins.This provides an experimen-tal basis for LbGP's potential role in supporting the im-provement of ovarian function.

Objective To observe the effects of wheat grain moxibustion on the hippocampal ERK/CREB-BDNF/TrkB signaling pathway and neurotransmitters in rats with perimenopausal depressive disorder(PDD);To explore its mechanism in treating PDD.Methods Totally 24 female SD rats were randomly divided into sham-operation group,model group,Western medicine group and wheat grain moxibustion group,with 6 rats in each group.The PDD rat model was established by bilateral ovariectomy and chronic unpredictable mild stress.The wheat grain moxibustion group received moxibustion at the"Zhongwan","Xiawan","Qihai","Guanyuan"acupoints,with a conical moxa cone for intervention,6 cones per acupoint;the Western medicine group was given fluoxetine hydrochloride solution 1.8 mg/kg by gavage,once a day,for consecutive 21 d.Before and after modeling and after intervention,the body mass,sucrose water preference rate and immobility time in the forced swimming test of the rats were recorded;Transmission electron microscopy was used to observe the synaptic structure in hippocampal tissue;ELISA was used to detect the expressions of 5-HT,GABA and Glu in hippocampal tissue;Western blot was used to detect the expressions of ERK,p-ERK,CREB,p-CREB,BDNF and TrkB proteins in hippocampal tissue.Results Compared with the sham-operation group,the body mass and sucrose water preference rate of the model group decreased,and the immobility time in the forced swimming test was prolonged(P<0.01);the number of synaptic vesicles in hippocampus decreased,the thickness of the postsynaptic density decreased,the aggregation of transduction-related proteins decreased,the synaptic cleft increased;the contents of 5-HT and GABA in hippocampal tissue decreased,the content of Glu increased,and the expressions of p-ERK,p-CREB,BDNF and TrkB proteins decreased(P<0.01).Compared with the model group,the body mass and sucrose water preference rate of the rats in Western medicine group and wheat grain moxibustion group increased,and the immobility time in the forced swimming test was shortened(P<0.01);the area of the postsynaptic density in the hippocampus increased,the synaptic cleft reduced,the contents of 5-HT and GABA in hippocampal tissue increased,the content of Glu decreased,and the expressions of p-ERK,p-CREB,BDNF and TrkB proteins increased(P<0.01).Conclusion Wheat grain moxibustion can effectively alleviate depressive-like behavior in PDD rats,and its mechanism may be related to regulating neurotransmitter expressions and mediating the ERK/CREB-BDNF/TrkB signaling pathway,thereby regulating hippocampal neuroplasticity.

Objective:To investigate the effect of Eriocitrin on the proliferation of esophageal cancer KYSE30 cells,and to explore its possible mechanism based on ferroptosis.Methods:Esophageal cancer KYSE30 cells were divided into 8 groups:the control group(conventional culture),RSL3 group(treated with 3 μmol/L ferroptosis inducer RSL3),Eriocitrin group(treated with 75 μmol/L Eriocitrin),Eriocitrin+Fer-1 group(treated with 5 μmol/L of ferroptosis inhibitor Fer-1 and 75 μmol/L Eriocitrin),Fer-1 group(treated with 5 μmol/L Fer-1 treatment),oe-NC group(transfected with blank vector control),oe-STAT3 group(transfected with STAT3 overexpression vector)and oe-STAT3+Eriocitrin group(transfected with STAT3 overexpression vector and then treated with 75 μmol/L Eriocitrin).Proliferation abilities of cells in each group were detected using CCK-8 assay,EdU incorporation assay and clone formation assay respectively.The levels of intracellular ferroptosis-related indicators were detected using the ELISA kits.Western blotting was used to detect the expression of STAT3/GPX4 pathway-related proteins.KYSE30 cell nude mouse subcutaneous transplanted tumor model was constructed to observe the effects of Eriocitrin and Fer-1 on the growth of transplanted tumors.Results:Eriodictyol could inhibit the proliferation and clone formation of KYSE30 cells,increase the levels of reactive oxygen species(ROS),malondialdehyde(MDA)and Fe2+,decrease the level of glutathione(GSH)(all P<0.05)and suppress the growth of transplanted tumors in nude mice.These effects could be reversed by Fer-1(P<0.05).Overexpression of STAT3 could abolish the inductive effect of eriodictyol on ferroptosis and its inhibitory effect on the STAT3/GPX4 pathway(P<0.05).Conclusion:Eriocitrin could induce ferroptosis in esophageal cancer KYSE30 cells by inhibiting STAT3/GPX4 signaling pathway and exert significant antitumor effects in esophageal cancer.

Objective:To investigate the prevalence of adult skeletal fluorosis caused by drinking tea-type endemic fluorosis in Yushu Tibetan Autonomous Prefecture (hereinafter referred to as Yushu Prefecture), Qinghai Province, and provide scientific basis for prevention and control of the disease.Methods:In August 2021, one village was selected as a survey site in six counties (cities) in Yushu Prefecture, including Nangqian, Chindu, Yushu, Zadoi, Qumarlêb, and Zhiduo. Drinking water samples and 10 brick tea samples were collected from each village to determine the fluoride content in water and brick tea; at least 100 permanent residents aged ≥ 25, who had a habit of drinking brick tea and had lived in the local area for more than 5 years, were selected for X-ray imaging to examine the prevalence of adult skeletal fluorosis.Results:A total of 75 samples of residential drinking water were collected, with a fluoride content of (0.21 ± 0.05) mg/L, ranging from 0.11 to 0.34 mg/L; 60 samples of brick tea, with a fluoride content of (626.70 ± 157.27) mg/kg, ranging from 324.00 to 2 102.00 mg/kg. A total of 1 136 adults were examined, and 318 cases of skeletal fluorosis were diagnosed, with a detection rate of 27.99%. Among them, the detection rates of mild, moderate, and severe skeletal fluorosis were 20.95% (238/1 136), 6.07% (69/1 136), and 0.97% (11/1 136), respectively, with mild symptoms being the main. The detection rates of skeletal fluorosis in males and females were 29.09% (121/416) and 27.36% (197/720), respectively, with no statistically significant difference between the gender (χ 2 = 0.39, P = 0.533). Comparison of the skeletal fluorosis in different gender, the differences were statistically significant (χ 2 = 22.31, P < 0.001). The detection rates of skeletal fluorosis in the age groups of 25 - 35, 36 - 45, 46 - 55, 56 - 65, 66 - 75, and ≥76 years old were 6.86% (7/102), 22.37% (51/228), 24.02% (92/383), 37.44% (73/195), 43.48% (70/161), and 37.31% (25/67), respectively. The differences between the groups were statistically significant (χ 2 = 59.84, P < 0.001). Moreover, there was a statistically significant difference in the composition of skeletal fluorosis among different age groups ( H = 37.66, P < 0.001). The Spearman correlation analysis results showed that the severity of adult skeletal fluorosis was positively correlated with age ( r = 0.34, P < 0.001). Conclusions:There is a certain degree of prevalence of adult skeletal fluorosis in Yushu Prefecture. And as age increases, the condition of skeletal fluorosis becomes more severe.

Objective To establish a high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)for the simultaneous determination of gefitinib,erlotinib,nillotinib and imatinib plasma concentrations and analyze the results.Methods The plasma samples were treated with acetonitrile precipitation and separated by Diamonsil C18 column(150 mm ×4.6 mm,3.5 μm)with mobile phase of 0.1％formic acid water(A)-0.1％formic acid acetonitrile(B).The flow rate of gradient elution was 0.7 mL·min-1,and the column temperature was 40 ℃ and the injection volume was 3 μL.Using arotinib as the internal standard,the scanning was carried out by using electrospray ionization source in positive ionization mode with multi-reaction monitoring.The specificity,standard curve,lower limit of quantitation,precision,accuracy,recovery rate,matrix effect and stability of the method were investigated.The concentrations of imatinib and erlotinib in 20 patients with chronic myelogenous leukemia(CML)and gefitinib and erlotinib in 3 patients with non-small cell lung cancer were measured.Results The standard curves of the four drugs were as follows,gefitinib:y=2.536 × 10-3x+9.362 × 10-3(linear range 20-2 000 ng·mL-1,R2=0.996 6);erlotinib:y=3.575× 10-3x+7.406 × 10-3(linear range 50-5 000 ng·mL-1,R2=0.994 9);nilotinib:y=1.945 x 10-3x+0.015 643(linear range 50-5 000 ng·mL-1,R2=0.990 6);imatinib:y=4.56 x 10-3x+0.010 451(linear range 100～104 ng·mL-1,R2=0.9963).RSD of intra-day and inter-day were less than 10％,and the accuracy ranged from 90％to 110％,and the recovery rates were 91.35％to 98.93％(RSD＜10％);the matrix effect ranged from 91.64％to 107.50％(RSD＜10％).Determination of 23 patients showed that the blood concentration of nilotinib ranged from 623.76 to 2 934.13 ng·mL-1,and the blood concentration of imatinib ranged from 757.77 to 2 637.71 ng·mL-1,and the blood concentration of gefitinib ranged from 214.76 to 387.40 ng·mL-1.The serum concentration of erlotinib was 569.57 ng·mL-1.Conclusion The method of this research is simple,fast,sensitive and dedicated,which can be monitored by the concentration of clinical blood.