OBJECTIVE To explore the effect and mechanism of warming channels and activating blood circulation external treat-ment to alleviate peripheral hyperalgesia in knee osteoarthritis(KOA)based on NGF/TrKA signaling pathway.METHODS 30 SD rats were randomly divided into normal group,KOA group and Yiceng group.KOA model was established by anterior cruciate ligament transection(ACLT).14 days after model establishment,rats in Yiceng group were treated with Yiceng patch.The peripheral pain threshold of rats was measured at different time points.The cartilage sections were stained with HE,Aggrecan and type II collagen.The synovial sections were stained with HE,Sirius red,silver and performed with immunostaining.The protein expression of key molecules NGF and TrKA of NGF/TrKA signaling pathway,inflammatory index IL-1β,pain mediator TRPV1,pan-neural mark-ers PGP9.5 and S100 in synovium and complexes transported to dorsal root ganglia(DRG)tissues via nerve endings was determined by Western Blot.The corresponding gene expression was determined by qPCR.The levels of NGF and SP in peripheral blood of rats were determined by ELISA.RESULTS Compared with the KOA group,the cold allodynia and mechanical allodynia thresholds of the rats in the Yiceng group increased(P<0.05,P<0.01);the protein and gene expression of NGF,TrKA,TRPV1,IL-1β,PGP9.5 in the synovial tissue decreased(P<0.05,P<0.01);the protein and gene expression levels of TRPV1,PGP9.5,S100 in the DRG tissue were downregulated(P<0.05,P<0.01).CONCLUSION The warming channels and activating blood circulation external treatment can inhibit the NGF/TrKA signaling pathway,downregulate the gene and protein expressions of NGF,TrKA,TRPV1,IL-1β,PGP9.5,and may inhibit the sprouting of sensory nerve fibers and improve the peripheral hyperalgesia state of rats with KOA.

Objective To analyze the current status and quality of existing obstetrics and gynecology enhanced recovery after surgery(ERAS)guidelines/consensus,understand the evidence-based practices in this field,and provide a basis for clinical decision-making.Methods A systematic search of PubMed,CNKI,and other databases,as well as NGC,SIGN,and websites of anesthesia societies domestically and internationally was conducted to obtain relevant guidelines and consensus on ERAS for gynecological surgery.The search was conducted from inception to July 1,2023,and the AGREE Ⅱ and RIGHT tools were used to analyze the current status and quality of the included guidelines/consensus,as well as to conduct a summary of the recommendations.Results A total of 5 guidelines and 10 consensus statements were included.The publication years were from 2018 to 2023,originating from the United States,China,Canada,and the United Kingdom.In the 6 areas evaluated using AGREEⅡ,clarity(66.06％)scored highest,followed by scope and purpose(56.67％),involvement of stakeholders(42.02％),and editorial independence(37.64％)with scores above 30％.The scores for rigor of development(26.88％)and applicability(4.17％)were relatively low.The same items among different evaluators indicated good consistency.The results of the scores in each area were as follows:A-level 2,B-level 13,and no C-level.Two high-quality guidelines were selected.The average RIGHT reporting rate was 44.38％.Subgroup analysis showed that guidelines were higher than consensus statements in the areas of rigor of development and applicability(P＜0.05),and that using GRADE was higher than not using(P＜0.05).The areas of rigor of development and clarity were higher in foreign guidelines than domestic guidelines(P＜0.05),and using evidence grading tools was higher than not using(P＜0.05).The summary of the recommendations showed that the quality of the evidence supporting the recommendations was uneven,and some recommendations were not specific enough.Conclusion The quality of the 15 included ERAS guidelines/consensus in obstetrics and gynecology is uneven,and 2 of them can be used as high-quality guidelines for clinical reference.The quality of evidence-based guidelines in foreign countries is higher,and the use rate of GRADE in foreign guidelines is high,most of which can provide clear recommendation strength and evidence level.Chinese guidelines/consensus are of lower quality compared to overseas,and methodology and report quality need further improvement.Overall,it is still necessary to strengthen the formulation of methods and reporting standards,improve the quality of evidence,and refine specific recommendations.

Objective To investigate the effect of Piezo1 activated by mechanical stress on knee osteoarthritis synovial fibrosis via the extracellular signal-regulated kinase(ERK)1/2 signaling pathway.Methods Twenty-five Sprague Dawley rats were divided into blank,exercise,exercise+GsMTx4,exercise+PD98059,and exercise+GsMTx4+PD98059 groups(n=5 per group).After modeling,serum and synovial tissue were extracted and collagen deposition was evaluated by Sirius red and Masson staining.Expression levels of Piezo1,ERK1/2,phospho(p)-ERK1/2,α-smooth muscle actin(SMA),transforming growth factor(TGF)-β,Collagen Ⅰ,and tissue inhibitor of metalloproteinase(TIMP)-1 were detected by Western blot and reverse transcription-quantitative polymerase chain reaction(RT-qPCR),and the interleukin(IL)-1β,IL-6,and tumor necrosis factor(TNF)-α contents were detected by enzyme-linked immunosorbent assay.For cell experiments,synovial cells were divided into blank,pull,pull+GsMTx4,pull+PD98059,and pull+GsMTx4+PD98059 groups and the above indices were detected in the model cells by Western blot,RT-qPCR,and other techniques.Results Mechanical stress increased collagen deposition in synovial tissues in the rats,and increased the protein and mRNA expression levels of the pathway-related and fibrosis-specific indicators Piezo1,p-ERK/ERK,α-SMA,TGF-β,Collagen I,and TIMP-1(P＜0.05).Piezo1 expression was significantly down-regulated by both inhibitors(P＜0.05),but the ERK inhibitor(PD98059)had no significant effect on Piezo1 gene expression.Levels of serum inflammatory factors were significantly higher in the exercise group compared with the blank group(P＜0.05),and levels were significantly reduced by the inhibitors(P＜0.05).The in vitro experiments showed the same trends as the animal experiments.Conclusions The Piezo1 ion channel can sense mechanical stress and activate the ERK 1/2 pathway to mediate knee synovial fibrosis.

Objective:To investigate the molecular expression and pathological features of endothelial cell (EC) in a murine model of choroidal neovascularization (CNV) based on single-cell RNA sequencing (scRNA-seq).Methods:Six C57BL/6 mice aged 6-8 weeks were randomly divided into two groups, with 3 mice in each group.Bilateral eyeballs were enucleated.The choroidal tissues from the two groups were isolated by shearing the complex and scraping the choroid, respectively.Single-cell suspension was prepared by continuous digestion with trypsin/type Ⅰ collagenase at 37 ℃, and the cell viability and EC ratio were detected by flow cytometry to determine the preparation method of single-cell suspension.Another 6 mice were randomly assigned into the control group and the CNV group, with 3 mice in each group.The CNV model was induced by laser photocoagulation and single-cell suspensions were prepared 7 days after modeling.Gene expression library construction was performed using the Chromi-um (10x Genomics) instrument.High throughput sequencing was performed using the Illumina Novaseq6000 to obtain the expression matrix.The EC subpopulations were classified according to previous researches and the Cellmarker database.Pseudo-time analysis was performed in EC, revealing the gene expression matrix of different states.CNV-EC were further selected with preliminary analysis of the expression characteristics.Another 6 mice were selected to establish the CNV model and eyeball frozen sections were prepared 7 days after modeling.Expression and distribution as well as the area percentage of EC marker Pecam1, mitochondrial outer membrane proteins Tomm20 and mt-Co1, and capillary markers Kdr and Plvap were observed by immunofluorescence staining, and the vascular diameter was calculated.The use and care of animals followed the ARVO statement.This study protocol was approved by the Experimental Animal Welfare and Ethics Committee of Air Force Military Medical University (No.20200181).Results:The cell viability of the single-cell suspension prepared from choroidal-scleral fragments and choroidal scrapings was 99.4% and 99.1%, respectively, both of which met the sequencing requirements.The percentage of EC detected by flow cytometry was approximately 1.58%.The scRNA-seq result revealed that both the normal control and CNV groups contained 13 choroidal cell clusters.Compared with the normal control group, the proportions of rod/cone photoreceptor cells, EC and hematopoietic cells all increased, while the retinal pigment epithelium (RPE) and Schwan cells reduced in the CNV group.Among all clusters, EC constituted 18.4%.The pseudo-time analysis demonstrated that EC could be further divided into 4 states.The percentage of state 2 EC was 29.1% in the CNV group, which was significantly higher than 9.5% in the normal control group.Differentially expressed gene analysis showed that the expression of mitochondrion-related genes, including mt-Nd4 and mt-Atp6, were upregulated in state 2 EC, while capillary-related genes, including Kdr and Esm1, were downregulated.Immunofluorescent staining revealed that the area of Tomm20 and mt-Co1 in Pecam1-positive EC in the CNV area was (19.50±4.68)% and (4.64±2.82)%, respectively, which were both higher than (3.00±2.09)% and (0.18±0.34)% in normal area ( t=7.88, 3.84; both at P<0.01). The area of Kdr and Plvap in Pecam1-positive EC in the CNV area was (1.50±0.29)% and (0.79±0.97)%, respectively, which were both lower than (31.30±5.44)% and (10.43±2.28)% in the normal area ( t=13.40, 9.48; both at P<0.01). The vascular diameter in the CNV area was (5.52±1.85)μm, which was larger than (4.21±1.84)μm in the normal area ( t=9.57, P<0.001). Conclusions:When CNV occurs, the proportion of EC in choroid increases, and CNV-EC shows pathologic features of mitochondrial metabolic activation and loss of capillary properties, suggesting the mitochondrial activation of EC may play a role in the formation of CNV.

Objective: To explore the intra-host genetic evolution of HIV-1 pol gene via follow-up for treatment-naïve HIV infections, and estimate the infection time with Bayesian coalescent theory, so as to support the evaluation of HIV epidemic. Methods: Five cases were recruited and followed up. The pol gene fragments were amplified for the characteristics of transmitted drug resistance ( TDR ) by RT-PCR. Bayesian coalescent theory was utilized to construct maximum clade credibility ( MCC ) tree for genetic evolution and calculate the time to the most recent common ancestor ( tMRCA ). Results: The five cases were all male, and aged from 27 to 50 years old.Five to nine sampling times were obtained from each case, and the pol gene sequences from each case formed a unique subcluster (posterior probability: 100% ), with different evolution characteristics, in the MCC tree. The three cases in primary HIV-1 infection were estimated to be infected one to five months before the first positive reaction of HIV screening, whereas the two HIV-1 diagnosed cases at first screening were extrapolated to get infected fourteen months and seven months before diagnosis, respectively. One case with acute HIV-1 infection carried TDR mutation ( M46I ) , expressing fast disease progress and quasispecies variation. Conclusions The general infection time can be estimated by analyzing the characteristics of intra-host genetic evolution of HIV-1 pol gene with Bayesian coalescent theory, and this method can help to estimate the HIV epidemic.

A double burden of overweight/obesity and malnutrition during childhood is a major concern in China. Dietary intakes in this critical period affect children's physical and cognitive development, and also have health consequences in later life. Therefore, establishing healthy eating habits that will endure is crucial for children. Nutrition education is an effective way in improving nutrition knowledge and attitudes, and healthy eating behaviors. Diverse forms of nutrition improvement programs that targeting children, family, teachers, and school settings have been conducted in many developed countries. However, due to the differences of genetic background, household environment as well as dietary patterns between Chinese children and children from other countries, the existing nutrition education programs for children abroad might not be appropriate for children in China. Thus, nutrition education programs that consider Chinese nutrition-related policies and food supply as well as the local educational resources are required for Chinese children. This review summarized nutrition-related policies and legislations in China and developed countries. A series of evidence-based nutrition education programs that combined educational strategies and environmental supports conducted in the Southwest China Childhood Nutrition and Growth Study were presented. These programs can serve as example models for adopting nutrition interventions to improve nutrition and health status of children in different regions of China.

Objective To evaluate the clinical effect of dextromethorphan and its effect on daily living of patients with poststroke pseudobulbar affect. Methods Sixty patients with poststroke pseudobulbar affect admitted in our hospital from May 2013 to October 2016 were enrolled. Then they were randomly divided into the control group and the treatment group,with 30 patients in each group.Patients in the control group were treated with fluox-etine therapy and patients in the treatment group were treated with dextromethorphan therapy.The center for neuro-logic study lability scale(CNS-LS)and activity of daily living(Barthel index,BI)before and 30 days after the treat-ments in the two groups had been accessed. Results Thirty days after the treatment,CNS-LS of the treatment group had obvious improvement compared with that before treatment(P < 0.01),but CNS-LS of the control group had no obvious improvement compared with that before treatment(P > 0.05). And significant improvement has been found 30 days after the treatment between the two groups(P<0.01).Furthermore,significant difference was found on BI between these two groups(P<0.05).Conclusions Dextromethorphan is effective in treatment of pa-tients with poststroke pseudobulbar affect and it can improve the activity of daily living of these patients.

Objective To observe the clinical effects of breaking blood expelling stasis method of traditional Chinese medicine (TCM) combined with edaravone on patients with acute hypertensive intra-cerebral hemorrhage and preliminarily discuss its protection mechanism on this disease. Methods Ninety-two patients with hypertensive intra-cerebral hemorrhage within 72 hours after occurrence admitted to Zengcheng District People's Hosipital of Guangzhou from May 2013 to December 2017 were enrolled, they were divided into conventional therapy group and combined treatment of TCM and western medicine group by random numbers produced by a computer, 46 cases in each group. The conventional therapy group was treated with conventional therapy, and the combined treatment of TCM and western medicine group was treated with conventional therapy, additionally the treatment of Poxue Zhuyu decoction (the ingredients of the decoction: leech 10 g, gradfly 10 g, rhubarb 15 g, cattail pollen 5 g, trichosanthes fruit 10 g, radix notoginseng 5 g, colla plastri testudinis 10 g, grassleaf sweetflag rhizome 10 g, ground beetle 3 g, dried lacquer 3 g, peach seed 10 g) and edaravone for 10 days. The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in serum were tested by enzyme-linked immunosorbent assay (ELISA) before and 7 days after treatment in the two groups; and the differences in National Institutes of Health stroke scale score (NIHSS) before and 14 days, 90 days after treatment as well as the changes of brain edema around the hematoma 14 days after treatment were compared between the two groups, and the incidence of adverse reaction was observed. Results After treatment, the levels of TNF-α, IL-6 and intra-cerebral hemorrhage quantities in the two groups were all decreased compared with those before treatment, and the degrees of decrease of TNF-α and cerebral hemorrhage volume in combined treatment of TCM and western medicine group were more significant than those in the conventional treatment group [TNF-α (ng/L): 21.00±6.10 vs. 29.40±11.33, cerebral hemorrhage volume (mL): 5.23±0.60 vs. 8.50±0.64, both P <0.05]. The IL-6 in the two groups were recovered to approximately normal levels after the treatment (ng/L: 13.60±5.36 vs. 15.40±6.13, P > 0.05). With the prolongation of therapeutic time, the scores of NIHSS of the two groups were significantly lowered than those before treatment, and the degree of reduction in the combined TCM and western medicine group on 90 days after treatment was more obvious than that of the conventional treatment group (4.34±0.67 vs. 7.73±0.61, P < 0.05). The volumes of edema around hematomas were increased after treatment in the two groups, but the degree of increase in combined treatment of TCM and western medicine group was slower than that of the conventional group (cm3: 7.57±0.64 vs. 10.16±0.60, P < 0.05). There was no statistical significant difference in the incidence of adverse reactions in comparison between the combined treatment of TCM and western medicine group and conventional therapy group [18.2% (8/44) vs. 20.5% (9/44), P > 0.05]. Conclusions Using breaking blood expelling stasis and edaravone for treatment of acute hypertensive intra-cerebral hemorrhage can accelerate the absorption of brain hematoma and improve the neurological function, and its mechanism may be relevant to the inhibition of some inflammatory factors.

Objective The aim of this study was to determine the content of phthalate in disposable plastic tableware sold on Chengdu market,and to provide primary data for safety evaluation.Methods Sample selection was based on stratified sampling.Sixteen phthalate compounds were investigated in 60 disposable plastic tableware,divided into seven groups.The analysis was performed by gas chromatography-mass spectrometry (GC-MS).Results In this survey,diethyl phthalate,diisobutyl phthalate,dibutyl phthalate and diethylhexyl phthalate were detected,while the other 12 phthalate compound were not.The positive rates of the four detected phthalate were 6.7％ (4/60),10.0％ (6/60),46.7％ (28/60) and 28.3％ (17/60) respectively,and the highest concentrations were 10.3,6.4,7.2 and 65.6 mg/kg,respectively.Conclusion The observed level of detection rates and maximum concentrations were relatively high in this survey.In addition,some subgroups of PAEs that were not allowed to use in food contact materials were detected.Therefore,the migration in different food simulant would be analyzed in the next step for further health outcome assessment.

Objective To study the determination method of total flavonoids in Gansu Astragali Radix and Hedysarum Polybotrys. Methods Calycosin glycosides etc. was selected as reference substances, comparison on the difference of absorption curves was done by ultraviolet spectroscopy and colorimetric method (NaNO2-Al(NO3)3-NaOH, AlCl3, Mg(Ac)2, NaOH, phosphomolybdic acid, HCl-Mg power). Results With colorimetric method, the maximum absorption wavelength of referrence and the test was inconsistent. The absorption peak shape was also different. With UV method, Calycosin glycosides in band Ⅱ (260 nm) showed a shoulder absorption. Astragali Radix and Hedysarum Polybotrys also showed characteristic shoulder absorptions in band Ⅱ with absorption wavelength at 263 nm and 265 nm. So the sample absorption wavelength is basically the same as that of the control sample. Conclusion Colorimetries usually used for determination of total flavonoids are not suitable for the comparison determination of Gansu Astragali Radix and Hedysarum Polybotrys. It is suitable for determining the contents of total flavonoids in samples by UV spectrophotometry at the band Ⅱ, which is the characteristic absorption band of isoflavone compound.