ObjectiveThis study aims to investigate the effect of Liuwei Dihuangwan on the autophagy function in the hippocampus of senescence-accelerated mouse prone 8 （SAMP8） by regulating the expression of glycoprotein non-metastatic melanoma protein B （GPNMB）. Furthermore， it is designed to explore the mechanism of the method of tonifying the kidneys and replenishing essence in the treatment of Alzheimer's disease （AD）. MethodsIn experiment 1， 24 5-month-old SAMP8 mice were randomly and equally divided into the model group， and the low-， middle- and high-dose（0.59，1.18，2.36 g·kg^-1） Liuwei Dihuangwan groups. At the same time， six 5-month-old senescence accelerated mouse resistant 1 （SAMR1） mice were used as the control group. The learning and memory ability was evaluated through novel object recognition experiment. Serum cortisol （Cort）， adrenocorticotropic hormone （ACTH） and urine 17-hydroxycorticosteroid （17-OHCS） levels were detected by enzyme-linked immunosorbent assay （ELISA）. The ultrastructure of hippocampal neurons was observed by transmission electron microscope （TEM）， and the expression levels of hippocampal GPNMB， a disintegrin and metalloproteinase 10 （ADAM10） and autophagy-related proteins were detected by Western blot. In experiment 2， 18 SAMP8 mice were randomly and equally divided into the model group， vector control group （Vector）， and GPNMB overexpression group （GPNMB^OE）. Lentiviral vectors were stereotactically injected into the brain （2 μL per side in the GPNMB^OE group）. Western blot was used to detect the expression of the above target proteins in the hippocampus； In Experiment 3， 24 SAMP8 mice were randomly and equally divided into the model group， Liuwei Dihuangwan group， Liuwei Dihuangwan+negative control （NC） group， and Liuwei Dihuangwan+GPNMB silencing group （shGPNMB）. Before drug treatment， the Liuwei Dihuangwan+NC group and the Liuwei Dihuangwan+shGPNMB group were injected with negative control and GPNMB silencing lentivirus， respectively. Western blot was used to detect the expression of the above target proteins in the hippocampus. ResultsThe novel object discrimination index of mice in the model group was significantly lower than that of mice in the control group （P<0.01）. The novel object discrimination index of mice in the medium- and high-dose Liuwei Dihuangwan groups was significantly higher than that of mice in the model group （P<0.01）. Aggregated autolysosomes were observed in the normal hippocampus tissue by TEM. In the model group， mitochondria were dominant， and no typical characteristic autophagosomes were observed. In the low- and medium-dose Liuwei Dihuangwan groups， a small number of autolysosomes and autophagosomes with double-membrane structures were observed. In the high-dose Liuwei Dihuangwan group， the number of autophagosomes and autolysosomes was greater than that in the low- and medium-dose groups. The results of ELISA and Western blot showed that compared with the control group， the levels of serum Cort， ACTH， and urine 17-OHCS in the model group were substantially increased， while the expression of hippocampal ADAM10， Beclin1， and microtubule associated-protein light chain 3-Ⅱ/Ⅰ （LC3 Ⅱ/Ⅰ） was significantly decreased. The expression of GPNMB and ubiquitin binding protein p62 was significantly increased （P<0.05， P<0.01）. Compared with the model group， the serum Cort and ACTH levels in the low-， medium-， and high-dose Liuwei Dihuangwan groups were significantly reduced， while only the urine 17-OHCS level in the high-dose group was significantly reduced. The hippocampal GPNMB， ADAM10， Beclin1， and LC3 Ⅱ/Ⅰ expression levels in the medium-， and high-dose groups of Liuwei Dihuangwan were significantly increased compared to the model group， whereas the expression of p62 was significantly reduced （P<0.01）. The above indicators showed a progressive trend among the three groups. Compared with the model group， the GPNMB^OE group showed a significant increase in GPNMB， ADAM10， Beclin1， LC3 Ⅱ/Ⅰ expression， and a significant decrease in p62 expression （P<0.01）. Compared with the model group， the expression of GPNMB， ADAM10， Beclin1， and LC3 Ⅱ/Ⅰ in the hippocampus of the Liuwei Dihuangwan group significantly increased， while the expression of p62 significantly decreased （P<0.01）. Compared with the Liuwei Dihuangwan group， the Liuwei Dihuangwan+shGPNMB group showed a significant decrease in GPNMB， ADAM10， Beclin1， LC3 Ⅱ/Ⅰ， and a significant increase in p62 expression （P<0.01）. ConclusionLiuwei Dihuangwan can enhance hippocampal autophagy function and improve AD by upregulating GPNMB expression.

Objective To retrospectively analyze the distribution and differences of oral behavior among patients with acute and chro-nic painassociated with temporomandibular disorders(TMD).Methods A total of 212 patients with pain TMD in West China Hospital of Stomatology completed oral behaviors checklist(OBC).SPSS statistical software was used to analyze all the data.Results The pa-tients with pain-related TMD had specific oral behaviors,such as"pressure on the jaw during sleep"(49.5%),"unilateral chewing"(49.5%),"upper and lower teeth contact when not eating"(37.7%),etc.Scores of"upper and lower teeth contact when not eating"and"muscle tension when teeth are not clenched or teeth are not in contact"in patients with chronic pain TMD were higher than those of acute patients,and the difference was statistically significant(P＜0.05).The total OBC scores and daytime OBC scores of chronic pain TMD patients were higher than those of acute pain patients,and the difference was statistically significant(P＜0.05).There was no significant difference in OBC scores of patients with acute and chronic pain TMD during sleep(P＞0.05).Conclusion Pain-related TMD patients had specific oral behaviors,and patients with chronic pain TMD exhibited higher levels of total OBC,daytime OBC,"upper and lower teeth contact when not eating"and"muscle tension when teeth do not bite or teeth are not in contact"oral behavior than acute pain TMD patients.In the treatment of patients with painful TMD,correcting the patient's bad oral behavior can help the treatment of pain-related TMD.

AIM:To investigate the effects of nebulized sodium tanshinone ⅡA sulfonate(STS)in a mouse model of pulmonary hypertension associated with chronic obstructive pulmonary disease(COPD-PH).METHODS:A to-tal of 32 healthy SPF-grade male C57BL/6 mice were randomly divided into 4 groups:control(CTL,n=8)group,COPD-PH(CS+LPS,n=8)group,STS-treated COPD-PH(CS+LPS+STS,n=8)group,and STS(n=8)group.The COPD-PH model was established through whole-body exposure to cigarette smoke(CS)combined with lipopolysaccharide(LPS)in-halation.Mice were subjected to cigarette smoke exposure in a chamber(9 cigarettes/h,2 h/session,2 sessions/d,6 d/week)for 60 d,except on days of LPS inhalation.On days 1 and 14,COPD-PH model mice received LPS(7.5 μg/mouse in 50 μL saline)via intranasal inhalation,while the CTL and STS groups received an equivalent volume of saline.STS was administered via nebulized inhalation(5 mg/kg,30 min per session,twice daily)immediately before CS exposure.At the end of the modeling period,lung function and right heart pressure were assessed.Bronchoalveolar lavage fluid(BALF)was collected for inflammatory cell counting.Levels of interleukin-6(IL-6)in BALF supernatants and plasma were measured using ELISA.Pathological changes in the airway and lung tissues were evaluated.RESULTS:(1)Com-pared to CTL mice,those exposed to CS and LPS exhibited lesions characteristic of COPD-PH,including emphysema,lung inflammation,decreased lung function,and increased right ventricular systolic pressure(RVSP)and right ventricu-lar hypertrophy index(RVHI)(P<0.05);(2)COPD-PH mice showed significantly elevated IL-6 levels in both BALF and plasma(P<0.05);(3)STS treatment alleviated emphysema and lung inflammation,improved lung function,prevent-ed increases in RVSP and the RV/(LV+S)ratio,and reduced IL-6 levels in both BALF and plasma(P<0.05).CON-CLUSION:The results indicate that nebulized inhalation of STS significantly slows the progression of COPD-PH,likely due to its ability to inhibit lung inflammation and reduce IL-6 expression in the lungs.

Objective To evaluate the impact of different exercise frequencies on the symptoms,quality of life,and duration of rehabilitative effects in patients with Parkinson's disease(PD).Methods A total of 90 PD patients(Hoehn and Yahr stage ≤3,receiving continuous dopaminer-gic stimulation)admitted in Department of Neurology of Chinese PLA General Hospital were en-rolled and randomly divided into groups G1,G2 and G3,with 30 participants in each group.They received rehabilitation exercises of Parkinson's Wellness Recovery(PWR)1 h per time,at differ-ent frequencies,that is,once(G1),twice(G2)and three times(G3)a week,for totally 6 months.Unified Parkinson's disease rating scale(UPDRS),levodopa equivalent dose,10-meter walk test(10-MWT),Fullerton advanced balance(FAB)Scale,trail making test-A(TMT-A),nine-hole peg test(9-HPT)and Parkinson's disease questionare-39(PDQ-39)were applied to evaluate the efficacy of the rehabilitation.Results The PDQ-39 score was significantly decreased in all the three groups after PWR exercise(P＜0.05,P＜0.01).Compared with the corresponding levels before intervention,the group G2 had obviously lower 10-MWT score and higher FAB score(P＜0.05),and the group G3 obtained notably decreased scores in UPDRS,UPDRS-Ⅲ,10-MWT,TMT-A,9-HPT,and PDQ-39[30.72±11.31 vs 31.41±10.42,P=0.001;19.21±9.72 vs 20.17±8.80,P=0.001;7.38(6.26,8.49)s vs 7.48(6.53,8.56)s,P=0.004;53.97±8.74 s vs 54.07±8.62 s,P=0.026;38.71(33.71,49.71)s vs 38.90(33.78,49.82)s,P=0.001;28.00(14.50,36.00)vs 30.00(15.50,38.50),P=0.001],and increased FAB score[23.00(16.00,33.00)vs 21.00(16.00,32.00),P=0.005].In addition,the patients in the G3 group experienced the most pronounced re-habilitation effect immediately after exercise,but,all aforementioned indicators remained augmen-ted even 4 weeks after exercise.Conclusion Exercise rehabilitation three times weekly can greatly enhance the motor function of PD patients,ameliorate non-motor symptoms,and the overall quali-ty of life,with the beneficial effects persisting for a minimum of 4 weeks.

Approximately 300 million tons of plastic are produced globally each year,which has a serious impact on human health,marine life and the livestock industry.Microplastics have also been detected in meat and milk samples.Research has shown that nanoplastics(NP)(＜1 μm)and mi-croplastics(MP)(1 μm-5 mm)can affect the digestive,immune and reproductive systems of ani-mals.This experiment aims to investigate whether NP and MP regulate autophagy and damage the nervous system and spatial memory of animals.This experiment was divided into control group,nanoplastic group(PS-NP group,0.1 μm)and microplastic group(PS-MP group,1 μm),with 20 mice in each group.The mice were given 0.5 mL of PS-NP and PS-MP every day for 35 consecutive days,followed by neck amputation and brain analysis.The results showed that NPs and MPs of dif-ferent diameters caused varying degrees of damage to the brains of mice.In the behavioral tests of new object recognition,barnes maze and Y-shaped maze spatial memory,compared with the control group,the PS-NP group and PS-MP group showed a significant decrease in spatial memory ability of mice.HE staining results showed that neuronal cells in the PS-NP and PS-MP groups of mice exhibited shrinkage,decreased cell volume and deepened staining.The number of Nissl bodies de-creased,leading to dissolution and disappearance.RT-PCR and Western blot results showed that compared with the control group,the expression of glutamate receptors NR1,NR2A and NR2B in-creased in mice administered NP and MP orally,while the expression of autophagy related proteins Parkin,LC3B and Beclin1 was inhibited.In summary,this study suggests that nanoplastics and mi-croplastics stimulate glutamate receptors in mice by inhibiting the autophagy pathway,leading to impaired spatial memory.

Objective:To elucidate the mechanism by which Clostridium butyricum alleviates atopic dermatitis (AD) from three aspects: immune cells, gut microbiota, and the metabolites of gut microbiota, short-chain fatty acids (SCFAs), and provide a theoretical reference for clinical probiotic-assisted treatment of AD. Methods:A model of 2, 4-dinitrochlorobenzene (DNCB)-induced AD was established using BALB/c mice. Three groups including control group, AD group, and Clostridium butyricum intervention group were set up with 20 mice in each group. The dermatitis score, scratching score, pathological conditions and mast cell infiltration at the lesion site, and the levels of cytokines related to Th1/Th2 and Th17/Treg as well as IgE levels in serum samples were analyzed. Gut microbiota was detected by 16S rRNA gene sequencing. The contents of SCFAs in mouse fecal samples were detected by gas chromatography-mass spectrometry. Spearman correlation analysis was performed to investigate the correlation between the cytokines related to Th1/Th2 and Th17/Treg, gut microbiota, and SCFAs. Comparisons between groups were performed using one-way analysis of variance with Turkey post hoc test correction. Results:Clostridium butyricum intervention down-regulated various inflammatory indexes and alleviated pathological changes in AD mice, elevated the levels of IFN-γ ( P<0.05) and IL-10 ( P<0.01), reduced the levels of IL-4, IL-17 and IgE ( P<0.01), and maintained the balance of Th1/Th2 ( P<0.01) and Th17/Treg ( P<0.001). Besides, the intervention improved intestinal dysbiosis by decreasing the abundance of conditionally pathogenic bacteria such as Prevotellaceae_ UCG-001 ( P<0.01) and increasing the abundance of beneficial bacteria such as Lachnospiraceae_ NK4A136_ group and norank_ f_ Oscillospiraceae ( P<0.05), and enhanced the production of SCFAs ( P<0.05). Correlation analysis showed that allergy-associated immune cytokines were strongly correlated with the composition of gut microbiota and the content of SCFAs. Conclusions:Clostridium butyricum may regulate the microbiota-SCFAs signaling response by inhibiting the colonization of harmful bacteria and increasing the abundance of beneficial bacteria. This, in turn, increases the level of SCFAs, decreases the number of pro-inflammatory cytokines, and maintains the balance of Th1/Th2 and Th17/Treg in the body. Therefore, Clostridium butyricum may alleviate allergic diseases.

Objective:To elucidate the mechanism by which Clostridium butyricum alleviates atopic dermatitis (AD) from three aspects: immune cells, gut microbiota, and the metabolites of gut microbiota, short-chain fatty acids (SCFAs), and provide a theoretical reference for clinical probiotic-assisted treatment of AD. Methods:A model of 2, 4-dinitrochlorobenzene (DNCB)-induced AD was established using BALB/c mice. Three groups including control group, AD group, and Clostridium butyricum intervention group were set up with 20 mice in each group. The dermatitis score, scratching score, pathological conditions and mast cell infiltration at the lesion site, and the levels of cytokines related to Th1/Th2 and Th17/Treg as well as IgE levels in serum samples were analyzed. Gut microbiota was detected by 16S rRNA gene sequencing. The contents of SCFAs in mouse fecal samples were detected by gas chromatography-mass spectrometry. Spearman correlation analysis was performed to investigate the correlation between the cytokines related to Th1/Th2 and Th17/Treg, gut microbiota, and SCFAs. Comparisons between groups were performed using one-way analysis of variance with Turkey post hoc test correction. Results:Clostridium butyricum intervention down-regulated various inflammatory indexes and alleviated pathological changes in AD mice, elevated the levels of IFN-γ ( P<0.05) and IL-10 ( P<0.01), reduced the levels of IL-4, IL-17 and IgE ( P<0.01), and maintained the balance of Th1/Th2 ( P<0.01) and Th17/Treg ( P<0.001). Besides, the intervention improved intestinal dysbiosis by decreasing the abundance of conditionally pathogenic bacteria such as Prevotellaceae_ UCG-001 ( P<0.01) and increasing the abundance of beneficial bacteria such as Lachnospiraceae_ NK4A136_ group and norank_ f_ Oscillospiraceae ( P<0.05), and enhanced the production of SCFAs ( P<0.05). Correlation analysis showed that allergy-associated immune cytokines were strongly correlated with the composition of gut microbiota and the content of SCFAs. Conclusions:Clostridium butyricum may regulate the microbiota-SCFAs signaling response by inhibiting the colonization of harmful bacteria and increasing the abundance of beneficial bacteria. This, in turn, increases the level of SCFAs, decreases the number of pro-inflammatory cytokines, and maintains the balance of Th1/Th2 and Th17/Treg in the body. Therefore, Clostridium butyricum may alleviate allergic diseases.

Objective Based on the"preventive treatment of disease"theory and real-world data,this study aimed to evaluate the clinical efficacy of Yangyin Shugan Formula in patients with ovarian dysfunction characterized by the syndrome of kidney deficiency and liver depression at different stages and to provide evidence for optimizing the timing of traditional Chinese medicine(TCM)in premature ovarian insufficiency(POI)management.Methods Ovarian dysfunction patients under 40 years old characterized by syndrome of kidney deficiency and liver depression,registered in the outpatient electronic medical records of the Second Affiliated Hospital of Guangzhou University of Chinese Medicine from January 2020 to December 2023,were stratified into three groups:subclinical POI,POI,and premature ovarian failure(POF)groups.The therapeutic outcomes of the three groups of patients were compared,including TCM syndrome total response rates,menstrual total response rates/normalization(cured)rates,base follicle-stimulating hormone(FSH)total response rate/normalization rates and anti-Müllerian hormone(AMH)levels.Results A total of 307 patients meeting the research criteria were included(97 cases in subclinical POI group,112 cases in POI group,and 98 cases in POF group).The TCM syndrome total response rates ranked as subclinical POI group(98.97%,96/97)>POI group(95.54%,107/112)>POF group(83.67%,82/98),with the POF group was lower than that of the subclinical POI and POI groups(P<0.017);while the difference of the subclinical POI group with the POI group was not significant.Menstrual total response rates ranked as subclinical POI group(97.22%,70/72)>POI group(72.32%,81/112)>POF group(51.02%,50/98),the differences among the three groups and in the post-hoc comparisons were significant(P<0.01,P<0.017);normalization rates ranked as subclinical POI group(90.28%,65/72)>POI group(54.46%,61/112)>POF group(26.53%,26/98),the differences among the three groups and in the post-hoc comparisons were significant(P<0.01,P<0.017).FSH total response rates ranked as subclinical POI group(89.69%,87/97)>POI group(81.25%,91/112)>POF group(60.20%,59/98),while POF group was lower than that of the other two groups(P<0.017),and the difference of the subclinical POI group with the POI group was not significant.Normalization FSH rates ranked as subclinical POI group(71.13%,69/97)>POI group(53.57%,60/112)>POF group(16.33%,16/98),the differences among the three groups and in the post-hoc comparisons were significant(P<0.01,P<0.017).Compared with before treatment,the FSH levels in all three groups decreased after treatment(P<0.01).The reduction of FSH in the POF and POI groups were better than that in the subclinical P OI group(P<0.017).No significant difference was observed in the reduction of FSH between the POI and POF groups.Post-treatment AMH elevation observed in the subclinical POI and POI groups(P<0.01).Conclusion Yangyin Shugan Formula improves the menstrual conditions and TCM syndromes across all the stages.It also improves elevated FSH levels across all the stages and has a specific effect on normalizing FSH and menstruation.Yangyin Shugan Formula increases the AMH levels of patients with subclinical POI and POI,highlighting the applicability of TCM in staged preventive management.

Objective:To understand the incidence of adverse pregnancy outcome in HIV-infected pregnant women and influencing factors in China and provide reference for the improvement of the health status of HIV-infected pregnant women and their newborns.Methods:Based on a mother-child cohort of HIV-infected pregnant women and children (PMTCT-MC-2005) established in Guangxi Zhuang Autonomous Region, Yunnan Province and Xinjiang Uygur Autonomous Region, this study enrolled pregnant women with or without HIV infection as study subjects from January 2017 to June 2023, a total of 1 646 pregnant women (558 HIV-infected and 1 088 HIV-uninfected) were included, and 34 cases with missing data were excluded. The χ2 test was used to analyze the difference in the incidence adverse pregnancy outcome between two groups, and used logistic regression model to identify the influencing factors of adverse pregnancy outcome in HIV-infected pregnant women. Results:A total of 1 612 pregnant women were included in the study, in whom 541 were infected with HIV and 1 071 were not infected with HIV. The incidence of adverse pregnancy outcome was 18.8% (303/1 612), the incidence of adverse pregnancy outcome was 33.1% (179/541) in the HIV-infected pregnant women and 11.6% (124/1 071) in the pregnant women without HIV infection. The results of multivariable logistic regression analysis showed that the influencing factors of adverse pregnancy outcome were age <35 years at delivery (a OR=0.64, 95% CI: 0.43-0.95) compared with the age ≥35 years and the duration of antiviral treatment over 10 years (a OR=0.43, 95% CI: 0.23-0.79) compared with less than one year. Conclusions:The incidence of adverse pregnancy outcome in HIV-infected pregnant women was high in some regions of China during 2017-2023. It is necessary for HIV-infected women to get pregnancy at appropriate time based on antiretroviral treatment effect and strengthen self-care to reduce the incidence of adverse pregnancy outcome.

Approximately 300 million tons of plastic are produced globally each year,which has a serious impact on human health,marine life and the livestock industry.Microplastics have also been detected in meat and milk samples.Research has shown that nanoplastics(NP)(＜1 μm)and mi-croplastics(MP)(1 μm-5 mm)can affect the digestive,immune and reproductive systems of ani-mals.This experiment aims to investigate whether NP and MP regulate autophagy and damage the nervous system and spatial memory of animals.This experiment was divided into control group,nanoplastic group(PS-NP group,0.1 μm)and microplastic group(PS-MP group,1 μm),with 20 mice in each group.The mice were given 0.5 mL of PS-NP and PS-MP every day for 35 consecutive days,followed by neck amputation and brain analysis.The results showed that NPs and MPs of dif-ferent diameters caused varying degrees of damage to the brains of mice.In the behavioral tests of new object recognition,barnes maze and Y-shaped maze spatial memory,compared with the control group,the PS-NP group and PS-MP group showed a significant decrease in spatial memory ability of mice.HE staining results showed that neuronal cells in the PS-NP and PS-MP groups of mice exhibited shrinkage,decreased cell volume and deepened staining.The number of Nissl bodies de-creased,leading to dissolution and disappearance.RT-PCR and Western blot results showed that compared with the control group,the expression of glutamate receptors NR1,NR2A and NR2B in-creased in mice administered NP and MP orally,while the expression of autophagy related proteins Parkin,LC3B and Beclin1 was inhibited.In summary,this study suggests that nanoplastics and mi-croplastics stimulate glutamate receptors in mice by inhibiting the autophagy pathway,leading to impaired spatial memory.