Objective: To investigate the effect of integrin α5 on the expression of NOD-like receptor thermal protein domain associated protein 3 (NLRP3) in periodontal ligament fibroblasts (PDLFs) within an inflammatory microenvironment. Methods: This study was approved by the Ethics Committee of Laboratory animals. After rat PDLFs were treated with LPS (0.5, 5, and 50 µg/mL) for 24 h, the primary medium was discarded and replaced with serum-free culture medium. After 24 h, the supernatant was collected and mixed with DMEM medium containing 10% exosome-free serum at a volume ratio of 1:1 to obtain conditioned medium (CM). The groups were labeled as the 0.5-CM, 5-CM, and 50-CM groups. In addition, PDLFs cultured in DMEM medium containing 10% exosome-free serum were considered the 0-CM group. PDLFs were cultured with the above CM. In the inhibitor group, PDLFs were cultured in 0-CM containing different concentrations of integrin α5 inhibitor ATN-161 (0, 0.025, 0.25, 2.5, 25, and 250 μg/mL). The effect of CM and integrin α5 inhibitor ATN-161 on cell viability was assessed using the CCK-8 assay. According to the CCK-8 results, in further inhibitor intervention experiments, PDLFs were cultured in 0-CM, 5-CM (without/with 25 μg/mL ATN-161), and 0-CM containing 25 μg/mL ATN-161, which were labeled as the 0-CM, 5-CM, ATN-161+5-CM, and ATN-161 groups, respectively. The expression changes of integrin α5 and NLRP3 were detected using Western blot and qRT-PCR techniques. For in vivo experiments, 24 rats were randomly divided into four groups (n=6). The control group contained healthy rats that received no treatment. The rats in the other three groups were injected with 40 µL of 0-CM containing 25 μg/mL ATN-161 or 5-CM (without or with 25 μg/mL ATN-161) on the palatal side of the left maxillary first molar every three days; these groups were classified as the ATN-161, 5-CM, and ATN-161+5-CM groups, respectively. On the 30th day, the left maxillary tissue of rats was used for Micro-CT, HE staining, and immunohistochemical detection. Results : The CCK-8 assay showed that CM, 25 μg/mL ATN-161, and ATN-161 concentrations below 25 μg/mL had no significant effect on cell viability at 12 h and 24 h (P > 0.05). 50-CM and 25 μg/mL ATN-161 significantly inhibited cell viability at 48 h (P < 0.05). For in vitro experiments, compared to the 0-CM group, both the protein and mRNA levels of integrin α5 and NLRP3 were significantly increased in rat PDLFs in the 5-CM group (P < 0.05). Intervention with 25 μg/mL ATN-161 significantly attenuated the enhancement of 5-CM on the expression of integrin α5 and NLRP3 (P < 0.05). For in vivo experiments, compared to the control group, alveolar bone resorption and periodontal inflammatory cell infiltration were significantly increased in the 5-CM and ATN-161+5-CM groups, and the expression of integrin α5 and NLRP3 was significantly increased (P < 0.01). However, compared to the 5-CM group, the ATN-161+5-CM group had less alveolar bone resorption and fewer periodontal inflammatory cells. Further, the expression of integrin α5 and NLRP3 was significantly reduced (P < 0.01). Conclusion In vitro and in vivo experiments showed that integrin α5 mediated NLRP3 expression in PDLFs under an inflammatory microenvironment. ATN-161 inhibited the expression of integrin α5, thus significantly downregulating the expression of NLRP3, which plays a role in inhibiting inflammation.

Objective To evaluate the efficacy of Jianpi Yang'er Ointment for children with anorexia of spleen deficiency type in Lingnan area;To analyze its effects on children's intestinal flora and functional information by means of metagenome sequencing.Methods Totolly 144 children with anorexia in Guangzhou Women and Children Medical Center of Guangzhou Medical University from August 2022 to January 2023 were orally treated with Jianpi Yang'er Ointment,one bag per time,twice a day,after meal,for 28 d.Visits were conducted at 0 and 28 days,respectively,to measure height and body mass,record symptoms and signs,observe adverse reactions,collect feces,and fill out the"Clinical Index Scoring Table for Children with Spleen Deficiency Type Anorexia in Lingnan Area".Clinically cured children were included in the anorexia group,with healthy children as a healthy control group(HC group),perform metagenomic analysis on feces of HC group and anorexia group 0 and 28 days of treatment.Results Compared with 0 d of treatment,there was no significant difference in the height of children(P>0.05),while there was a significant difference in body mass(P<0.05),the total syndrome score,main symptom score,and secondary symptom score decreased at 28 d of treatment(P<0.05).The efficacy index was 57.03%,and the total effective rate was 99.31%(143/144).All patients did not experience symptoms such as allergies,vomiting,or diarrhea.7 clinically cured children and 6 healthy children were included in metagenome analysis on feces.PCoA showed that the structure of intestinal flora in anorexia group with 28 d treatment(JP2 group)was more similar to that in HC group at the phylum and genus levels;at the phylum level,there was no significant difference in the distribution of intestinal flora between anorexia group with 0 d treatment(JP1)group and JP2 group and HC group.At the genus level,compared with JP1 group,the abundance of Bacteroides in JP2 group decreased,but the difference was not statistically significant(P>0.05);at the species level,both JP1 and JP2 groups were mainly composed of Phocaeicola plebeius et al.and Bifidobacterium pseudocatenatum et al.The biomarker with significant differences between the two groups was s_Phascolarctobacterium_faecium,while the HC group was mainly composed of Prevotella copri and Bifidobacterium pseudocatenatum.The functional annotation results indicated that a significant change in microbial functional genes related to carbohydrate metabolism occurred after the treatment with Jianpi Yang'er Ointment.Conclusion Jianpi Yang'er Ointment can effectively improve the clinical symptoms of children with spleen deficiency anorexia in Lingnan area,adjust the structure of intestinal flora,increase the content of key probiotics,and regulate the production of intestinal microbiota short-chain fatty acids.

Objective:To investigate the outcomes of periacetabular osteotomy (PAO) through modified Smith-Petersen or Bikini approach with an auxiliary posterolateral incision for developmental dysplasia of hip (DDH).Methods:85 patients (97 hips) who underwent PAO through modified Smith-Petersen or Bikini approach with auxiliary posterolateral incision for DDH in the 920th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army or the Second Xiangya Hospital of Central South University from January 2016 to January 2021 were retrospectively analyzed. There were 70 females and 15 males, with an average age of 28.6±8.4 years (12-49). According to the Hartofilakidis classification, all patients were classified as type Ⅰ. There were 77 hips classified as Grade 0 and 20 hips classified as Grade 1 according to the T?nnis classification. The X-ray evaluation including the lateral central edge angle (LCEA), t?nnis angle (TNS) and anterior central edge angle (ACEA), visual analogue score (VAS) and modified Harries hip score (mHHS) were recorded preoperatively and during each postoperative follow-up. The cumulative sum (CUSUM) method was used to fit the learning curves of operation time.Results:All patients were followed-up with an average of 52.7±18.9 months (21 to 84). The average operative time and intraoperative blood loss were 95.9±28.2 min (65 to 215) and 414.7±97.0 ml (250 to 900), respectively. The learning curve of operation time was 10 cases. The average LCEA 32.39° (30.29°, 34.92°), TNS 3.14° (1.56°, 5.67°) and ACEA 31.55° (29.07°, 33.88°) were all significantly improved compared to preoperative values ( Z=-30.764, P<0.001; Z=30.595, P<0.001; Z=-38.134, P<0.001). The average VAS was reduced from 5.18±0.88 points preoperatively to 1.27±0.93 points postoperatively with significant difference ( t=51.231, P<0.001). The average mHHS was increased from 73.12±9.17 to 92.02±4.05 postoperatively with significant difference ( t=-26.902, P<0.001). No patients received total hip arthroplasty during the follow-up period. Conclusion:Bernese PAO through modified Smith-Petersen or Bikini approach with a small auxiliary posterolateral incision was not only a safe and effective method for the treatment of DDH but also shortened the learning curve and reduced difficulty of the surgery.

Objective:To observe any effect of ultrashortwave (USW) therapy on inflammatory cytokines and the MAPK pathway of rats with a spinal cord injury.Methods:Seventy-nine Sprague-Dawley rats were randomly divided into a control group ( n=35), an intervention group ( n=35) and a sham group ( n=9). Allen′s method was used to establish a contusion model of SCI in the rats of the control and intervention groups, while the sham group′s spinal tissues were exposed but not stricken. Beginning twenty-four hours after SCI modeling, the intervention group was given 7min of USW therapy daily, five days a week till the day of sacrifice for sampling the target area of spinal cord for tests. Then, motion function was evaluated using Basso, Beattie and Bresnahan (BBB) scoring. One, three and seven days after the SCI modeling, immunofluorescence and western blotting were employed to observe any changes in inflammatory factors and the MAPK pathway in the lesioned area. Results:Fourteen days after the modeling the average BBB score of the intervention group was significantly higher than the control group′s average. Moreover, 7 days after the modeling the average content of the domains containing protein 3 (NLRP3), interleukin-6 (IL-6), IL-6 receptor and tumor necrosis factor-α (TNF-α) in the target area of the spinal cord of sham group showed significantly lower levels than in the other 2 groups. And the levels in the intervention group were significantly lower than in the control group. Seven days after the modeling the number of cells positive for zinc finger protein 36 (TTP) in the lesioned area of the intervention group was significantly greater than among the control group. At the same time the levels of MAPK-activated protein kinase 2 (MK2), phosphorylated-mitogen-activated protein kinase-activated version (p-MK2) and TTP in the control and intervention groups were significantly higher than in the sham group. And there were significant differences between the intervention group and control group in the levels of MK2, p-MK2 and TTP.Conclusion:Ultrashortwave therapy can inhibit inflammation by regulating the MAPK inflammatory pathway, promoting the recovery of motion functions, at least in rats.

Objective:To describe the characteristics and change trends of incidence, mortality and disease burden of acute myocardial infarction (AMI) in Qingdao from 2014 to 2020.Methods:We analyzed the incidence data of AMI retrieved from Qingdao Chronic Diseases Surveillance System. The average annual percent change (AAPC) of morbidity and mortality of AMI were evaluated by using Joinpoint log-linear regression model. Disability adjusted life year (DALY) was used to estimate disease burden of AMI in Qingdao.Results:A total of 70 491 AMI cases and 50 832 deaths of AMI occurred in Qingdao from 2014 to 2020. The age-standardized morbidity and mortality were 54.71/100 000 and 36.55/100 000, respectively. During 2014-2020, the AAPC of age-standardized morbidity was 2.86% (95% CI: 0.42%-5.35%), and 4.30% (95% CI: 1.24%-7.45%) in men and 0.78% (95% CI: -0.89%-2.47%) in women, respectively. The log-linear regression model showed that age-standardized morbidity in age groups 30-39, 40-49 years increased rapidly, with the AAPCs of 8.92% (95% CI: 2.23%-16.06%) and 6.32% (95% CI: 3.30%-9.44%), respectively. The trend was also observed in age groups 30-39, 40-49 and 50-59 years in men, with the AAPCs of 11.25% (95% CI: 3.54%-19.54%), 6.73% (95% CI: 2.63%-10.99%) and 6.72% (95% CI: 2.98%-10.60%), respectively. There was no significant change in age-standardized mortality. The DALY rate increased from 7.49/1 000 in 2014 to 8.61/1 000 in 2020, with the AAPC of 1.97% (95% CI: 0.36%-3.60%). Conclusions:The age-standardized morbidity of AMI in men increased in Qingdao, especially in those aged 30-49 years, while age-standardized mortality rate of AMI was relatively stable from 2014 to 2020. The burden of disease of AMI increased in both men and women.

Objective:To explore the effect of repetitive transcranial magnetic stimulation (rTMS) combined with cognitive behavioral therapies (CBT) on the cognitive function and alcohol craving in patients with alcohol dependence.Methods:From March 2019 to September 2021, a total of 150 patients with alcohol dependence were enrolled and randomly divided into rTMS treatment group (rTMS+ sham CBT, n=41), CBT treatment group (CBT+ sham rTMS, n=34), rTMS+ CBT treatment group( n=36) and control group (sham rTMS+ sham CBT, n=39). At baseline (before treatment), 2nd week, 8th week, 12th week and 24th week, alcohol dependence scale (ADS) was used to evaluate the degree of alcohol dependence, the obsessive compulsive drinking scale (OCDS) was used to assess patients' drinking craving, and Montreal cognitive assessment scale (MoCA) was used to assess the overall cognitive level of patients.SPSS 23.0 statistical software was used to compare the differences of ADS, OCDS and MoCA scale scores of the four groups by repeated measure ANOVA and simple effect analysis. Results:(1)The patients in the four groups were evaluated with ADS scale at baseline, 12th week and 24th week respectively.The interaction of group×time( F=1.279, P=0.279) and the main effect of group were not significant ( F=0.882, P=0.454), and the main effect of time was significant ( F=12.925, P<0.001) .Further simple effect analysis showed that the ADS score of rTMS+ CBT group was lower than that of baseline(14.48±5.70, 10.00±6.51) ( P=0.01) at 24th week.(2)Patients in the four groups were assessed with OCDs scale at baseline, 2nd week, 8th week, 12th week and 24th week, and the interaction of group×time was significant ( F=2.015, P=0.042). Further simple effect analysis showed that the OCDs scores of rTMS group and rTMS+ CBT group at each follow-up time node were lower than those at baseline period (all P<0.05). (3)Patients in the four groups were assessed with MoCA scale at baseline, 8th week, 12th week and 24th week, and the interaction of group×time was not significant ( F=1.660, P=0.106), and the main effect of group and the main effect of time were significant ( F=2.964, P=0.038; F=14.239, P<0.001). Further simple effect analysis showed that the score of MoCA scale in CBT group at the 24th week was higher than that at baseline (21.73±5.81, 24.60±3.98)( P=0.029), the score of MoCA scale in rTMS+ CBT group at the 24th week was higher than that at the 8th week (23.50±6.01, 25.95±2.87) ( P=0.006), and the score of MoCA scale in rTMS group at the 12th week was higher than that in control group (22.08±6.64, 26.64±2.46)( P=0.009). Conclusion:rTMS combined with CBT can be effective in improving alcohol craving and cognitive function in patients with alcohol dependence, and has a good long-term effect.

Objective:To investigate the protective effect of external diaphragm pacing on the prevention of ventilator-induced diaphragm dysfunction (VIDD) in rabbits and its mechanism.Methods:Eighty-five New Zealand white rabbits were randomly (random number) divided into the blank control group (BC, n=5), spontaneous breathing group (SB, n=20), volume control ventilation group (VC, n=20), external diaphragm pacing group (EDP, n=20), external diaphragm pacing and volume control ventilation group (EDP+ VC, n=20). After successful modeling, the rabbits in each group were treated accordingly except for the BC group. Rabbitss in the BC group were not mechanically ventilated, and the diaphragm was removed immediately after anesthetizing. Whole diaphragms of 5 rabbits per time point per other group were also collected after anesthesia at post treatment hour (PTH) 6 and on post treatment day (PTD) 1, 3, and 7. Diaphragm weight/body weight and diaphragm isometric contractile force of each group were measured. The pathological changes of diaphragmatic tissues were observed by HE staining. The protein expressions of Cyt c, RyR1, caspase-3, and p-mTORC1 were measured by Western blot. Repeated measures analysis of variance was used for the comparison between multiple groups of variables at different time points, and LSD- t test was used for the further comparison between two groups at the same time point, a P<0.05 was considered statistically significant. Results:Compared with the BC group, the VC group showed diaphragmatic pathological changes conformed to VIDD: DW/BW was decreased obviously; HE staining revealed obvious changes in diaphragmatic tissue; Diaphragmatic contractility was also significantly decreased; The expression of Cyt c and caspase-3 were increased while the expression of RyR1 and p-mTORC1 were decreased gradually with the extension of treatment time ( P<0.05). Compared the EDP+VC group with the VC group, with the extension of treatment time, DW, DW/BW, pathological damages and diaphragmatic contractility were improved [PTD 1: (0.80±0.05)kg vs (0.56±0.04) kg, PTD 3: (1.06±0.05) kg vs (0.47±0.03) kg, PTD 7: (1.24±0.10) kg vs (0.39±0.07) kg, all P<0.05; PTD 1: (2.05±0.54) vs (1.86±0.72), PTD 3: (2.19±0.61) vs (1.74±0.40), PTD 7: (2.46±0.62) vs (1.53±0.85), all P<0.05; PTD 1: (2.39±0.42) N/cm 2vs (1.91±0.25) N/cm 2, PTD 3: (2.57±0.62) N/cm 2vs (1.72±0.50) N/cm 2, PTD 7: (2.77±0.55) N/cm 2vs (1.54±0.33) N/cm 2, all P<0.05]. The expression of Cyt c and caspase-3 were decreased while the expression of RyR1 and p-mTORC1 were increased gradually in the EDP+VC group ( P<0.05). Conclusions:External diaphragm pacer plays a protective role in ventilator-induced diaphragm dysfunction, which can inhibit mitochondrial damage, reduce oxidative damage, and mitigate diaphragmatic atrophy and injury.

Objectives:To examine the prognosis factors of recurrence of esophageal carcinoma within 6 months after neoadjuvant therapy followd by surgery.Methods:The clinical data of 187 patients with esophageal squamous cell carcinoma who underwent neoadjuvant therapy followed by curative esophagectomy between January 2018 and April 2020 at Department of Thoracic Surgery, Shanghai Chest Hospital were analyzed retrospectively. There were 160 males and 27 females, aging (63.0±7.1) years (range:43 to 76 years). The t test, χ 2 test and rank-sum test were used for univariate analysis of the prognosis factors for recurrence within 6 months postoperative, while the Logistic regression was used for multivariate analysis. Results:There were 30 patients (16.0%) developed recurrence within 6 months after operation, including local recurrence in 1 case, regional recurrence in 11 cases, hematogenous recurrence in 13 cases, and combined recurrence in 5 cases. Univariate analysis suggested that there was a significant difference in T staging of tumor before neoadjuvant therapy (cT), tumor regression grade, circumferential resection margin, pathological T stage (ypT) and pathological N stage (ypN) between the recurrence patients and non-recurrence patients (all P<0.05). Logistic regression analysis suggested that the cT3-4 ( OR=2.701, 95% CI: 1.161 to 6.329, P=0.021) and ypN(+)( OR=1.654, 95% CI: 1.045 to 2.591, P=0.032) were the independent prognosis factors for recurrence within 6 months. Conclusion:The combination of neoadjuvant therapy and surgery is not effective in reducing early postoperative recurrence in patients who have invaded the epineurium before treatment, and still have positive lymph nodes after neoadjuvant therapy.

Objectives:To examine the prognosis factors of recurrence of esophageal carcinoma within 6 months after neoadjuvant therapy followd by surgery.Methods:The clinical data of 187 patients with esophageal squamous cell carcinoma who underwent neoadjuvant therapy followed by curative esophagectomy between January 2018 and April 2020 at Department of Thoracic Surgery, Shanghai Chest Hospital were analyzed retrospectively. There were 160 males and 27 females, aging (63.0±7.1) years (range:43 to 76 years). The t test, χ 2 test and rank-sum test were used for univariate analysis of the prognosis factors for recurrence within 6 months postoperative, while the Logistic regression was used for multivariate analysis. Results:There were 30 patients (16.0%) developed recurrence within 6 months after operation, including local recurrence in 1 case, regional recurrence in 11 cases, hematogenous recurrence in 13 cases, and combined recurrence in 5 cases. Univariate analysis suggested that there was a significant difference in T staging of tumor before neoadjuvant therapy (cT), tumor regression grade, circumferential resection margin, pathological T stage (ypT) and pathological N stage (ypN) between the recurrence patients and non-recurrence patients (all P<0.05). Logistic regression analysis suggested that the cT3-4 ( OR=2.701, 95% CI: 1.161 to 6.329, P=0.021) and ypN(+)( OR=1.654, 95% CI: 1.045 to 2.591, P=0.032) were the independent prognosis factors for recurrence within 6 months. Conclusion:The combination of neoadjuvant therapy and surgery is not effective in reducing early postoperative recurrence in patients who have invaded the epineurium before treatment, and still have positive lymph nodes after neoadjuvant therapy.

Ischemic stroke is a severe disorder resulting from acute cerebral thrombosis. Here we demonstrated that post-ischemic treatment with ciclopirox olamine (CPX), a potent antifungal clinical drug, alleviated brain infarction, neurological deficits and brain edema in a classic rat model of ischemic stroke. Single dose post-ischemic administration of CPX provided a long-lasting neuroprotective effect, which can be further enhanced by multiple doses administration of CPX. CPX also effectively reversed ischemia-induced neuronal loss, glial activation as well as blood-brain barrier (BBB) damage. Employing quantitative phosphoproteomic analysis, 130 phosphosites in 122 proteins were identified to be significantly regulated by CPX treatment in oxygen glucose deprivation (OGD)-exposed SH-SY5Y cells, which revealed that phosphokinases and cell cycle-related phosphoproteins were largely influenced. Subsequently, we demonstrated that CPX markedly enhanced the AKT (protein kinase B, PKB/AKT) and GSK3 (glycogen synthase kinase 3) phosphorylation in OGD-exposed SH-SY5Y cells, and regulated the cell cycle progression and nitric oxide (NO) release in lipopolysaccharide (LPS)-induced BV-2 cells, which may contribute to its ameliorative effects against ischemia-associated neuronal death and microglial inflammation. Our study suggests that CPX could be a promising compound to reduce multiple ischemic injuries; however, further studies will be needed to clarify the molecular mechanisms involved.