Objective:To investigate damaging effects of clomifene citrate(CC)on endometrial stromal cells(hEndoSCs),and to study effects of spermidine on autophagy and inflammatory cytokine expression in damaged endometrial stromal cells.Methods:Groups were firstly divided into control group,spermidine group,clomiphene group(CC group),CC+Spermidine group.MTT assay was used to detect cell survival rate of hEndoSCs after co-incubation with different concentrations of CC or Spermidine for 24 h.Con-tent of intracellular reactive oxygen species(ROS)and level of apoptosis in cells of the 4 groups were detected by flow cytometry tech-nique.Western blot was used to detect expressions of autophagy pathway-related proteins ULK1,p-ULK1,LC-3Ⅱ,and apoptosis-re-lated proteins Bax,Bcl-2,Cleaved-caspase 3.RT-qPCR was used to detect mRNA expressions of IL-6,IL-1β and TNF-α.Results:Compared with control group,CC group showed decreased cell survival,increased apoptosis rate,ROS content,Bax,Cleaved-cas-pase 3 expressions,decreased Bcl-2 expression,decreased levels of autophagy-related proteins p-ULK1 and LC-3Ⅱ/Ⅰ,and elevated expressions of inflammatory factors IL-6,IL-1β and TNF-α mRNA(P<0.01).There was no significant changes viability of cells in spermidine group compared with control group(P>0.05).Compared with CC group,cell survival rate in CC+spermidine group was sig-nificantly increased,apoptosis rate,ROS content,Bax and Cleaved-caspase 3 expressions were decreased,Bcl-2 expression was in-creased,expressions of autophagy-related proteins p-ULK1 and LC-3Ⅱ/Ⅰ were elevated,while expressions of inflammatory factors IL-6,IL-1β and TNF-α mRNA were decreased(P<0.01).Conclusion:CC can inhibit endometrial stromal cell proliferation,promote apoptosis,and increase the transcript levels of inflammatory factors IL-6,IL-1β and TNF-α.Spermidine can reduce intracellular ROS in clomiphene-injured endometrial stromal cells by activating cellular autophagy,increase cell survival,and inhibit the expressions of inflammatory factors IL-6,IL-1β and TNF-α.

Objective:To discuss the effect of Wnt/β-catenin signaling pathway inhibitor methyl 3-｛[(4-methyl-phenyl)sulfonyl]amino ｝ benzoate(MS AB)on the fibrogenic response of the human endometrial stromal cells(HESCs),and to provide the foundation for the application of MSAB in the target therapy of intrauteriue adhesion(JUA).Methods:The normal HESCs were cultured in vitro and divided into two groups:control group and transforming growth factor β1(TGF-β1)group;the HESCs from the adhesion part of the IUA patients were cultured in vitro,regarded as IUA group.Western blotting method was used to detect the expression levels of fibrotic marker protein type Ⅰ collagen α1(COL1A1)in the cells in various groups at different time points(0,12,24,48,and 60 h)after treated with TGF-β1.MTT assay was used to detect the proliferation activities of the cells in various groups.Western blotting method was used to detect the expression levels of the fibrotic marker protein COL1A1,stromal marker proteins such as N-cadherin and α-smooth muscle actin(α-SMA),and Wnt/β-catenin signaling pathway-related protein β-catenin in the cells in control and IUA groups.Based on the MSAB concentrations,the normal HESCs were divided into 0(control),0.25,0.50,0.75,and 1.00 μmol·L-1 MSAB groups,and MTT assay was used to detect the survival rates of the cells in various groups.After treated with MSAB,the normal HESCs were divided into control group(normal HESCs),TGF-β1 group(10 μg·L-1 TGF-β1 induced normal HESCs for 24 h then the drug was withdrawn,replaced with complete culture medium,and the cells continued to be cultured for 24 h),and MSAB group(10 μg·L-1 TGF-β1 induced normal HESCs for 24 h then the drug was withdrawn,replaced with a complete medium containing 0.75 μmol·L-1 MSAB and the cells continued to be cultured for 24 h).Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of epithelial-mesenchymal transition(EMT)-related transcription factors Snail,Slug,Smuc,ZEB1,and ZEB2,and COL1A1 mRNA in the cells in various groups.Western blotting method was used to detect the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in various groups.Results:Compared with control group(after treated with TGF-β1 for 0 h),the expression levels of COL1A1 proteins in the HESCs after treated with TGF-β1 for 12,24,48,and 60 h in TGF-β1 group were increased(P＜0.05 or P＜0.01).Compared with control group,there was no significant difference in the proliferation activity of the HESCs in IUA and TGF-β1 groups(P＞0.05).Compared with control group,the expression levels of COL1A1,β-catenin,N-cadherin,and α-SMA proteins in the cells in IUA group were increased(P＜0.05 or P＜0.01).Compared with control group,the survival rates of the cells in 0.75 and 1.00 μmol·L-1 MSAB groups were decreased(P＜0.05 or P＜0.01).Compared with control group,the expression levels of Snail,Slug,and COL1A1 mRNA in the cells in TGF-β1 group were increased(P＜0.05 or P＜0.01);compared with TGF-β1 group,the expression levels of Snail,Slug,and COL1A1 mRNA in the cells in MSAB group were decreased(P＜0.05 or P＜0.01).Compared with control group,after treated with TGF-β1 for 24 h,the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in TGF-β1 group were increased(P＜0.01);compared with TGF-β1 group,the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in MSAB group were decreased(P＜0.05 or P＜0.01).Conclusion:MSAB can inhibit the fibrogenic responses of the HESCs in vitro,and the results provide the theoretical basis for the application of MSAB in the target therapy of IUA.

Objective:To investigate the effects of bisphenol A(BPA)on the proliferation activity and stemness characteristics of endometrial mesenchymal stem/stromal cells(eMSCs),and to elucidate the improvement effect of human umbilical cord mesenchymal stem cell-derived supernatant(hUCMSC-Sup)on the cell injury.Methods:The eMSCs were cultured in vitro and treated with different concentrations of BPA(0,200,250,300,350,and 400 μmol·L-1).The eMSCs were divided into control group(only cultured with culture solution),BPA group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA),BPA+hUCMSC-Sup group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA and 50%volumetric ratio of hUCMSC-Sup),and BPA+CHIR-99021 group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA and 10 μmol·L-1 CHIR-99021).The survival rates of eMSCs in various groups were detected by methyl thiazolyl tetrazolium(MTT)assay.The numbers and diameters of the spheroids in various groups were detected by spheroids formation assay,the proliferation activities of the cells in eMSCs stem cell spheroids in various groups were detected by CCK-8 assay;the percentage of CD73+cells in eMSCs in various groups were detected by flow cytometry;the expression levels of sex determining region Y-box 2(Sox2),octamer-binding transcription factor 4(Oct4),and Nanog mRNA in the eMSCs in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)method,the expression levels of β-catenin protein in the eMSCs in various groups were detected by Western blotting method.Results:The MTT results showed that after treated with BPA for 24 and 48 h,compared with 0 μmol·L-1 BPA group,the survival rates of eMSCs in 200,250,300,350,and 400 μmol·L-1 BPA groups were significantly decreased(P<0.01).At 24 and 48 h after treatment,compared with control group,the survival rate of the eMSCs in BPA group was significantly decreased(P<0.01);at 48 h after treatment,compared with BPA group,the survival rate of the eMSCs in BPA+hUCMSC-Sup group was significantly inereased(P<0.05).The spheroids formation assay results showed that compared with culture 3 d group,the numbers and diameters of stem cell spheroids of the eMSCs in culture 4 d group and culture 5 d group were significantly increased(P<0.05 or P<0.01);compared with control group,after 48 h of culture,the number and diameter of the cells in eMSCs stem cell spheroids in BPA group were significantly decreased(P<0.05 or P<0.01).The CCK-8 results showed that after 24 and 48 h of treatment,compared with control group,the proliferation activity of the cells in eMSCs stem cell spheroids in BPA group was significantly decreased(P<0.01);compared with BPA group,the proliferation activity of the cells in eMSCs stem cell spheroids in BPA+hUCMSC-Sup group was significantly increased(P<0.01).The flow cytometry results showed that compared with control group,the percentage of the CD73+cells in the eMSCs in BPA group was significantly decreased(P<0.01);compared with BPA group,the percentage of the CD73+cells in eMSCs in BPA+hUCMSC-Sup group was significantly increased(P<0.01).The RT-qPCR results showed that compared with control group,the expression levels of Sox2,Oct4,and Nanog mRNA in the cells in BPA group were significantly decreased(P<0.01);compared with BPA group,the expression levels of Sox2,Oct4,and Nanog mRNA in the cells in BPA+hUCMSC-Sup group and BPA+CHIR-99021 group were significantly increased(P<0.01).The Western blotting results showed that compared with control group,the expression level of β-catenin protein in the eMSCs in BPA group was significantly decreased(P<0.01);compared with BPA group,the expression levels of β-catenin protein in the eMSCs in BPA+hUCMSC-Sup group and BPA+CHIR-99021 group were signifrcantly inereased(P<0.01).Conclusion:BPA can inhibit the stemness characteristics of the eMSCs,and injury the self-renewal and repair of endometrium;its mechanism may be related to down-regulating the activity of Wnt/β-catenin signal pathway in the cells.hUCMSC-Sup can promote the proliferation of injured eMSCs,and has improvement effect on the stemness injury induced by BPA.

This article reported a survived case of amniotic band syndrome (ABS) following fetal reduction by radiofrequency ablation. The woman conceived monochorionic diamniotic twin pregnancy spontaneously. Prenatal ultrasound at 24 weeks of gestation indicated twin-twin transfusion syndrome (stage Ⅲ), and radiofrequency ablation for fetal reduction was successfully performed after formal consent. At 28 +6 weeks, ultrasound reexamination revealed significant edema in the left foot of the fetus, with banding around the ankle, as well as the strangulation mark and narrowing rings. Fetal ABS (ⅡB stage) was diagnosed after multidisciplinary consultation. An immediate emergency cesarean section was performed and a live male baby was born. A thin amniotic band could be seen wrapping around the left ankle of the newborn for several rounds, with obvious strangulation marks about 1 cm deep into the skin, and significant edema on the dorsum and sole of the foot, and the submalleolus area. The amniotic band was released at once, and the edema faded gradually after surgery. After a follow-up of 28 days, the lower limbs of the newborn became normal.

BACKGROUND: The clinical features of enthesitis in patients with psoriatic arthritis (PsA) have been reported in some Western countries, but data in China are very limited. This study aimed to describe the characteristics of enthesitis in Chinese patients with PsA and compared them with those in other cohorts. METHODS: Patients with PsA enrolled in the Chinese Registry of Psoriatic Arthritis (CREPAR) (December 2018 to June 2021) were included. Data including demographics, clinical characteristics, disease activity measures, and treatment were collected at enrollment. Enthesitis was assessed by the Spondyloarthritis Research Consortium of Canada (SPARCC), Maastricht ankylosing spondylitis enthesitis score (MASES), and Leeds enthesitis index (LEI) indices. A multivariable logistic model was used to identify factors related to enthesitis. We also compared our results with those of other cohorts. RESULTS: In total, 1074 PsA patients were included, 308 (28.7%) of whom had enthesitis. The average number of enthesitis was 3.3 ± 2.8 (range: 1.0-18.0). More than half of the patients (165, 53.6%) had one or two tender entheseal sites. Patients with enthesitis had an earlier age of onset for both psoriasis and arthritis, reported a higher proportion of PsA duration over 5 years, and had a higher percentage of axial involvement and greater disease activity. Multivariable logistic regression showed that axial involvement (odds ratio [OR] 2.21, 95% confidence interval [CI], 1.59-3.08; P <0.001), psoriasis area and severity index (PASI) (OR: 1.03, 95% CI: 1.01-1.04; P = 0.002), and disease activity score 28-C reactive protein (DAS28-CRP) (OR: 1.25, 95% CI: 1.01-1.55; P = 0.037) were associated with enthesitis. Compared with the results of other studies, Chinese patients with enthesitis had a younger age, lower body mass index (BMI), a higher rate of positive human leukocyte antigen (HLA)-B27, more frequent dactylitis, and a higher proportion of conventional synthetic disease-modifying antirheumatic drugs' (csDMARDs) use. CONCLUSIONS Enthesitis is a common condition among Chinese patients with PsA. It is important to evaluate entheses in both peripheral and axial sites.
Humans ; Arthritis, Psoriatic/drug therapy* ; East Asian People ; Enthesopathy/complications* ; Registries ; Severity of Illness Index ; Spondylarthritis/epidemiology*

Objective To analyze the individualized CTV-to-PTV margin dose and positioning errors in radiotherapy for esophageal cancer for improving the treatment accuracy while meeting dose requirements.Methods Fifty-four esophageal cancer patients admitted to Sun Yat-sen University Cancer Center at Huangpu District from June 2021 to June 2022 were enrolled.All of the patients underwent CBCT scans in each fraction,and a total of 1283 CBCT images were collected.The image registration between CBCT image before radiotherapy and planning CT image was carried out to obtain errors in vertical(VRT),longitudinal(LNG),lateral(LAT),Roll,Pitch,and YAW directions.The mean values of six-dimensional positioning errors in the first 5 fractions were calculated,and the results were compared with the total fractional errors using the single sample t-test method for determining the differences.The CTV-to-PTV margin was calculated with the formula(margin=2.5∑+0.7δ),and the calculated margins were divided into 5 groups:Group A(5 mm expansion in all directions),Group B(7.9 mm expansion in LAT direction,and 5 mm expansion in other directions),Group C(11.03 mm expansion in LNG direction,and 5 mm expansion in the other directions),Group D(6.36 mm expansion in VRT direction,and 5 mm expansion in the other directions),and Group E(7.9 mm expansion in LAT direction,11.03 mm expansion in LNG direction,and 6.36 mm expansion in VRT direction).Simulation planning was conducted for 10 patients.Results The proportions of differences between the mean values of six-dimensional errors in the first 5 fractions and the total fractional errors in 54 patients were analyzed.There was no significant difference in 192 out of the 324 directions in 54 patients,accounting for 59.26%(P>0.05).Among them,the LAT,LNG,VRT,Pitch,Roll and YAW directions accounted for 64.81%,57.41%,51.85%,64.81%,57.41%and 59.26%of the total cases.The calculated CTV-to-PTV margin was 7.90,11.03 and 6.36 mm in LAT,LNG and VRT directions.The statistical analysis showed that the differences in the coverage rates of organs-at-risk and target areas among the 5 groups of CTV-to-PTV margins were trivial(P>0.05).Conclusion Using the positioning errors in the first 5 fractions of radiotherapy for esophageal cancer to predict subsequent positioning errors is feasible.The reasonable individualized margin in radiotherapy for esophageal cancer can reduce the inter-fractional off-target rate without increasing the dose delivered to organs-at-risk.The study provides a reference for the target volume margin of esophageal cancer and an important basis for precision treatment.

Objective:To investigate the effect of total alkaloid of harmaline (TAH) on inducing cellular autophagy and degradating of neurotoxic proteins Tau and α-synuclein (α-Syn).Methods:(1) The in vitro cultured PC12 cells were divided into blank control group, and 1, 2.5, 5, 10, 20 and 50 μg/mL TAH groups, respectively; and they were treated with 0, 1, 2.5, 5, 10, 20 and 50 μg/mL TAH for 24 h; cell morphology and number were observed, and cell survival rate was determined by MTT assay. (2) PC12 cells were divided into blank control group, rapamycin group, and 1, 2.5, 5, 10 and 20 μg/mL TAH groups; these cells were treated with same amount of solvent, 50 nmol/L autophagy activator rapamycin, and 1, 2.5, 5, 10 and 20 μg/mL TAH for 4 h, respectively, and the number of autophagosomes was detected by immunofluorescent staining. (3) PC12 cells were divided into blank control group, rapamycin group, and 10 μg/mL TAH group; these cells were treated with same amount of solvent, 50 nmol/L rapamycin, and 10 μg/mL TAH for 4 h; the protein expression levels of p62 and microtubule-associated protein 1 light chain 3 II (LC3-II) was detected by Western blotting. (4) PC12 cells were divided into blank control group, chloroquine group, TAH group, and TAH+chloroquine group; these PC12 cells were treated with 50 nmol/L autophagy inhibitor chloroquine, 10 μg/mL TAH, and 10 μg/mL TAH+50 nmol/L chloroquine for 4 h, respectively; the LC3-II protein expression was detected by Western blotting. (5) PC12 cells were divided into TAH group and blank control group; 10 μg/mL TAH and same amount of solvent were given to each group for 4 h, and then, the phosphorylated mammalian target of rapamycin (p-mTOR) and phosphorylated 70-KD ribosomal protein S6 kinase (p-P70S6K) protein expression levels were detected by Western blotting. (6) Tet on HEK293 cells with Tau-green fluorescent protein (GFP) overexpression were divided into blank control group, TAH group, doxycycline group, doxycycline+TAH group, doxycycline+TAH+3-MA group, and doxycycline+TAH+chloroquine group. Cells in the later 4 groups were treated with 200 ng/mL Tet system inducer doxycycline for 24 h; cells in the blank control group were treated with same amount of solvent, those in the TAH group were treated with 10 μg/mL TAH, and cells in the latter 3 groups were treated with 10 μg/mL TAH, 10 μg/mL TAH+5 mmol/L 3-MA, and 10 μg/mL TAH+50 nmol/L chloroquine, respectively, for 24 h; the changes of green fluorescence intensity of these cells were observed under laser confocal microscope. The Tau-GFP and LC3-II protein expression levels were detected by Western blotting. (7) HEK293 cells with stable α-Syn expression were divided into blank control group, chloroquine group, TAH group and TAH+chloroquine group; these cells were treated with same amount of solvent, 50 nmol/L chloroquine, 10 μg/mL TAH and 10 μg/mL TAH+50 nmol/L chloroquine for 24 h, respectively; the α-Syn and LC3-II protein expression levels were detected by Western blotting. Results:(1) As compared with that in the blank control group, the cell survival rate in 20 and 50 μg/mL TAH groups was significantly lower, and that in the 50 μg/mL TAH group was statistically lower than that in 20 μg/mL TAH group ( P<0.05). (2) As compared with that in the blank control group, the number of autophagosomes in rapamycin group, and 10 and 20 μg/mL TAH groups was significantly increased, and that in 10 μg/mL TAH group was statistically higher than that in 20 μg/mL TAH group ( P<0.05); 10 μg/mL TAH group was selected for subsequent experiments. (3) As compared with the blank control group, the rapamycin group and TAH group had significantly decreased P62 protein expression and significantly increased LC3-II protein expression ( P<0.05). (4) As compared with that in the blank control group, the LC3-II protein expression in the chloroquine group, TAH group and TAH+chloroquine group was significantly increased, and LC3-II protein expression in TAH+chloroquine group was statistically higher than that in chloroquine group ( P<0.05). (5) The p-mTOR and p-p70S6K expression levels in the TAH group were significantly decreased as compared with those in the blank control group ( P<0.05). (6) The Tau-GFP protein expression in doxycycline group was significantly increased as compared with that in the blank control group ( P<0.05); that in doxycycline+TAH group was significantly decreased as compared with that in the doxycycline group ( P<0.05); that in the doxycycline+TAH+3-MA group and doxycycline+TAH+chloroquine group was statistically increased as compared with that in doxycycline+TAH group ( P<0.05). The LC3-II protein expression in the TAH group was significantly increased as compared with that in the control group, that in the doxycycline+TAH group was significantly increased as compared with that in the doxycycline group, that in the doxycycline+TAH+3-MA group was significantly decreased as compared with that in the doxycycline+TAH group, and that in doxycycline+TAH+ chloroquine group was significantly increased as compared with that in the doxycycline+TAH group ( P<0.05). Conclusion:TAH may activate autophagy by inhibiting the mTOR/p70S6K signaling pathway, which in turn promotes the degradation of neurotoxic proteins Tau and α-Syn.

Objective To analyze the effects of different supplements on anemia related indexes in rural children. Methods A stratified method was adopted, and six villages (towns) in and around Qinghai Province were selected as intervention sits for the present study. A total of 304 children from 2 to 6 years old at each intervention site meeting the inclusion criteria were screened and divided into three groups (A, B, and C), who were intervened for 3 months. Serum vitamin A, vitamin D and hemoglobin levels were measured before and after the intervention. Results The hemoglobin level of 304 children before intervention was (118.65±16.07) g /L, and the prevalence of anemia was 9.54%. The vitamin A value, vitamin D value and hemoglobin value were increased after three months of the intervention. The changes of vitamin A value, vitamin D value and hemoglobin value in rural children in group C were significantly higher than those in groups A and B. The increase in vitamin A value in rural children aged 3 years was significantly higher than that in other age groups, and the increase in hemoglobin in rural children of 1 year old was significantly higher than that in other age groups. The increase in vitamin A value of rural children of other ethnic groups (mainly Tibetans) was significantly higher than that of Han and Hui nationalities, and the increase of hemoglobin value in Hui rural children was significantly higher than that in Han and other ethnic groups. Conclusion Vitamin A combined with iron dextran tablets was effective in preventing anemia in rural children.

Objective: To explore the clinical effect of the multi-point, multi-level suspension as well as fixation using midfacial soft-tissue spaces for midface lifting. Methods: A total of 65 patients with aging midface were admitted at the First Affiliated Hospital of Fujian Medical University from October 2017 to February 2019. Among them, 47 patients underwent primary blepharoplasty and midface lifting. Eighteen patients, including 5 patients with lower eyelid retraction or ectropion after blepharoplasty, underwent secondary midface lifting after blepharoplasty. The preseptal space was separated under orbicularis oculi muscle by palpebral margin incision. The orbicularis retaining ligament and the tear trough ligament were severed through preperiosteal plane. The preseptal space was connected with premaxillary space and prezygomatic space. The malar fat pad and superficial fascia were vertically suspended and fixed on the periosteum of infraorbital ridge by selected medial, middle and lateral points. The orbicularis oculi muscle was suspended and superolaterally fixed at lateral orbital periosteum. Therefore, the midface could be lifted by multi-point, multi-level suspension and fixation. Results: All incision healed in the first stage. Eyelid separation occurred to 1 patient, around 1 month after the operation. Tarsal strip lateral canthoplasty was performed for repair. Local protuberance of lateral lower eyelid occurred to another patient shortly after the operation, but improved after 3 months by lid massaging. No other complication was observed in the rest of the cases. All patients were followed up for 1 to 8 months and the results were satisfactory. Conclusions It is simple and practicable to utilize midfacial soft-tissue spaces. This method could benefit patient of less trauma, bleeding, and complications, and good clinical effect. It is a good choice for rejuvenation of the midface, especially for secondary midface rejuvenation after blepharoplasty, or complicated with lower eyelid retraction and ectropion.

Objective:To probe the implement of inquiry -based teaching in nursing ethics. Method:105 nursing students attended the innovation of inquiry-based teaching for Nursing Ethics. Results:88. 5% nursing students satisfied with innovation of teaching;82 . 9% nursing studentsethics awareness was enhanced largely in understanding for nursing ethics;68 . 6% nursing students considered that their ability for ethical thinking has been enhanced largely;93 . 3% nurs-ing students thought that nursing ethics was important. Different inquiry methods for nursing students led to different level of analysis ability for ethic cases(P﹤0. 001). Conclusions:It's necessary to perform the innovation of inquiry-based teaching, promoting the student's activity, focus on learning process and enhancing the students comprehensive ability.