Objective To observe the clinical efficacy of adjuvant PD-1 inhibitor in the treatment of patients with hepatocellular carcinoma after microwave ablation and the changes of lymphocyte subsets before and after treatment.Methods A total of 56 patients with hepatocellular carcinoma who underwent microwave ablation in our hospital were randomly divided into two groups according to different treatments,with 28 cases in each group.Patients in the group A received adjuvant treatment with PD-1 inhibitors after microwave ablation,while patients in the group B received microwave ablation alone.The local efficacy of microwave ablation,the survival of patients after adjuvant treatment of PD-1 inhibitor and the occurrence of related adverse reactions were observed.Peripheral venous blood was collected to detect the number of T lymphocyte subsets before microwave ablation in the two groups,after microwave ablation in the group B,and after six cycles of immunotherapy in the group A.Results There was no significant difference in the complete ablation rate of cases between group A and group B(89.3% vs.85.7%,P=0.69),and there was no significant difference in the complete ablation rate of lesions between group A and group B(91.7% vs.85.3%,P=0.40).The median progression-free survival(mPFS)of the group A was not yet reached,the mPFS of group B was 12.6 months,and there was a statistically significant difference in the mPFS between the two groups(P=0.03).The median overall survival(mOS)of the two group were not yet reached.The proportion of CD4+T lymphocytes after treatment in the group A significantly increased compared with that in the group B[(37.05±2.22)% vs.(29.23±2.88)%,P=0.03],while the proportion of CD8+T lymphocytes[(21.08±2.23)% vs.(27.18±1.75)%,P=0.04)]and the proportion of Treg cells[(4.26±0.31)% vs.(5.22±0.34)%,P=0.04]significantly decreased.There was no serious adverse reactions in all patients after microwave ablation,and 28 patients in the group A had no adverse reactions above grade 3 after PD-1 inhibitor treatment.Conclusion Adjuvant PD-1 inhibitor therapy after microwave ablation can prolong the progression-free survival of patients,enhance the function of T lymphocytes,and weaken the role of immunosuppressive cells,which may be an effective strategy to delay the recurrence of liver cancer after microwave ablation.

ObjectiveThis study aimed to observe the impact of sinomenine hydrochloride on the proliferation of fibroblasts and the mRNA expression of related genes in knee joint adhesion and contracture in rabbits. Additionally, we sought to explore its potential mechanisms in combating knee joint adhesion and contracture. MethodsFibroblasts were cultured in vitro, and experimental groups with varying concentrations of sinomenine hydrochloride were established alongside a control group. Cell proliferation was assessed using the CCK-8 assay. Changes in the mRNA expression of fibroblast-related genes following sinomenine hydrochloride treatment were evaluated using RT-qPCR. The impact of the drug on serum levels of inflammatory cytokines was determined using the ELISA method, and the expression of related proteins was assessed using Western blot. ResultsSinomenine hydrochloride was found to inhibit fibroblast viability, with viability decreasing as the concentration of sinomenine hydrochloride increased. The effects of sinomenine hydrochloride in all experimental groups were highly significant (P<0.05). At the mRNA expression level, compared to the control group, sinomenine hydrochloride led to a significant downregulation of inflammatory cytokines in all groups (P<0.05). Additionally, the expression levels of apoptosis-related proteins significantly increased, while Bcl-2 mRNA expression decreased (P<0.05). The mRNA expression levels of the PI3K/mTOR/AKT3 signaling pathway also decreased (P<0.05). At the protein expression level, in comparison to the control group, the levels of inflammatory cytokines IL-6, IL-8, IL-1β, and TGF-β were significantly downregulated in the middle and high-dose sinomenine hydrochloride groups (P<0.05). The expression levels of cleaved-PARP, cleaved caspase-3/7, and Bax increased and were positively correlated with the dose, while the expression levels of the anti-apoptotic protein Bcl-2 and the PI3K/AKT3/mTOR signaling pathway were negatively correlated with the dose. Sinomenine hydrochloride exhibited a significant inhibitory effect on the viability of rabbit knee joint fibroblasts, which may be associated with the downregulation of inflammatory cytokines IL-6, IL-8, and IL-1β, promotion of apoptosis-related proteins cleaved-PARP, cleaved caspase-3/7, and Bax, suppression of Bcl-2 expression, and inhibition of gene expression in the downstream PI3K/AKT3/mTOR signaling pathway. ConclusionSinomenine hydrochloride can inhibit the inflammatory response of fibroblasts in adhesive knee joints and accelerate fibroblast apoptosis. This mechanism may offer a novel approach to improving and treating knee joint adhesion.

Objective To explore the protective mechanism of RNA binding protein LIN28 on diabetic nephropathy(DN).Methods LIN28 was overexpressed or knocked down by adenovirus in HEK 293T cells.The gene and functional signaling pathway significantly changed after overexpression of LIN28 were obtained through RNA Seq transcriptome sequencing and bioinformatics analysis.HEK 293T cells were divided into the Control group,the Treatment group,the Adv-LIN28-OE+D-ribose group and the Adv-LIN28-NC+D-ribose group in in vitro LIN28 overexpression studies.And HEK 293T cells were divided into the Control group,the Treatment group,the Adv-shRNA LIN28+D-ribose group and the Adv-shRNA NT+D-ribose group in in vitro LIN28 knockdown studies.ELISA was used to detect the levels of human advanced glycation end products(AGEs)of cells supernatants in the above groups.Western blot was used to detect the expression levels of RAGE,NF-κB and MMP-2 of cells in the above groups.Results The results of RNA-Seq transcriptome sequencing,GO analysis and KEGG analysis showed that overexpression of LIN28 resulted in a significant down-regulation of AGE-RAGE signaling pathway in HEK 293T cells(P<0.05).The results of ELISA showed that compared with the Control group,the cell in the Treatment group produced a large amount of AGEs(P<0.05);compared with the Treatment group,the AGEs level of cells in the Adv-LIN28-OE+D-ribose group was significantly decreased(P<0.05),while the AGEs level of cells in the Adv-shRNA LIN28+D-ribose group was increased(P>0.05).The results of Western blot showed that compared with the Control group,the expression levels of RAGE,NF-κB and MMP-2 of cells in the Treatment group were significantly increased(P<0.05);compared with the Treatment group,the expression levels of RAGE,NF-κB and MMP-2 of cells in the Adv-LIN28-OE+D-ribose group were significantly decreased(P<0.05),while the expression levels of RAGE,NF-κB and MMP-2 of cells in the Adv-shRNA LIN28+D-ribose group were significantly increased(P>0.05).Conclusion Overexpression of LIN28 by adenovirus at the cellular level in vitro can lead to significant differential expression of thousands of genes.In particular,it can inhibit the diabetes and complications-related AGE-RAGE signaling pathway,which is critical in the progression of DN disease,and play a role in protecting DN.

Objective: BMI may play a protective role in reducing the mortality rate of patients with chronic obstructive pulmonary disease (COPD), but its effect on acute exacerbation of COPD remain unclear. Methods: Subjects were selected from the COPD patients registration system established in 2014 in Pudong new district, Shanghai. COPD patients from 8 communities were selected by cluster sampling and follow up was conducted prospectively for 18 months. Basic information and BMI were obtained from baseline survey, and acute exacerbations were collected during follow-up. The association between BMI and risk of acute exacerbation was evaluated by using multiple negative binomial regression. Results: Among 328 community COPD patients, 295 who completed the follow up were included in the analysis, in whom 96.3% (284/295) were mild COPD patients. During the follow-up, 11.1% (33/295) of the patients reported acute exacerbation. The results of multiple negative binomial regression suggested that, the risk for acute exacerbation decreased with the increase of BMI (IRR=0.85, 95%CI:0.73-0.98), overweight patients with BMI ≥25.0 kg/m² (IRR=0.36, 95%CI:0.13-0.91) or moderate BMI (T₂ vs. T₁, IRR=0.31, 95%CI:0.11-0.77) had lower risk for acute exacerbation compared with the patients with normal or low BMI. BMI had a linear correlation with the risk of acute exacerbation. Conclusion: The risk for acute exacerbation in patients with mild or moderate COPD in communities decreased with the increase of BMI, and being overweight might be a protective factor for the acute exacerbation of COPD.
Body Mass Index ; China/epidemiology* ; Disease Progression ; Humans ; Overweight/complications* ; Prospective Studies ; Pulmonary Disease, Chronic Obstructive/complications*

The vagina contains at least a billion microbial cells,dominated by lactobacilli.Here we perform metagenomic shotgun sequencing on cervical and fecal samples from a cohort of 516 Chinese women of reproductive age,as well as cervical,fecal,and salivary samples from a second cohort of 632 women.Factors such as pregnancy history,delivery history,cesarean section,and breastfeeding were all more important than menstrual cycle in shaping the microbiome,and such information would be necessary before trying to interpret differences between vagino-cervical micro-biome data.Greater proportion of Bifidobacterium breve was seen with older age at sexual debut.The relative abundance of lactobacilli especially Lactobacillus crispatus was negatively associated with pregnancy history.Potential markers for lack of menstrual regularity,heavy flow,dysmenor-rhea,and contraceptives were also identified.Lactobacilli were rare during breastfeeding or post-menopause.Other features such as mood fluctuations and facial speckles could potentially be predicted from the vagino-cervical microbiome.Gut and salivary microbiomes,plasma vitamins,metals,amino acids,and hormones showed associations with the vagino-cervical microbiome.Our results offer an unprecedented glimpse into the microbiota of the female reproductive tract and call for international collaborations to better understand its long-term health impact other than in the settings of infection or pre-term birth.

Objective:To investigate the effect of resveratrol on the expression of inflammatory cytokines and related genes in human SZ95 sebocytes induced by benzo (a) pyrene.Methods:Human SZ95 sebocytes were cultured in vitro, and divided into 4 groups: control group treated with 1‰ dimethyl sulfoxide for 27 hours, resveratrol group treated with 1 × 10 -5 mol/L resveratrol for 24 hours, benzo (a) pyrene group treated with 1 × 10 -5 mol/L benzo (a) pyrene for 3 hours, resveratrol+benzo (a) pyrene group treated with 1 × 10 -5 mol/L resveratrol for 24 hours followed by 1 × 10 -5 mol/L benzo (a) pyrene for 3 hours. Real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of interleukin (IL) -1α, IL-6, aryl hydrocarbon receptor (AhR) , cytochrome P4501A1 (CYP1A1) and cytochrome P4501B1 (CYP1B1) in SZ95 sebocytes in the above groups; Western blot analysis was conducted to determine the phosphorylation level of p38 mitogen-activated protein kinase (p38 MAPK, expressed as the ratio of phosphorylated to total p38 MAPK) and AhR protein expression; enzyme-linked immunosorbent assay (ELISA) was conducted to detect levels of IL-1α and IL-6 in the cell culture supernatant in each group. One-way analysis of variance was used for comparison of means among multiple groups, and least significant difference- t test was used for multiple comparisons. Results:The mRNA and protein expression of IL-1α in SZ95 sebocytes significantly differed among the control group, resveratrol group, benzo (a) pyrene group and resveratrol+benzo (a) pyrene group (mRNA: 2.045 ± 0.272, 2.058 ± 0.154, 3.124 ± 0.094, 2.185 ± 0.337, protein: 9.132 ± 1.181, 9.429 ± 0.771, 20.361 ± 0.907, 9.917 ± 0.897, F=14.662, 101.705, P < 0.01, < 0.001, respectively) , and were significantly lower in the resveratrol+benzo (a) pyrene group than in the benzo (a) pyrene group (both P < 0.01) . In addition, the phosphorylation level of p38 was significantly higher in the benzo (a) pyrene group than in the control group, resveratrol group and resveratrol+benzo (a) pyrene group ( F=303.129, P < 0.000 1) . The mRNA expression of AhR, CYP1A1 and CYP1B1 was significantly lower in the resveratrol+benzo (a) pyrene group than in the benzo (a) pyrene group ( t=10.64, 33.599, 18.327, respectively, all P < 0.001) . The benzo (a) pyrene group showed significantly decreased protein expression of AhR compared with the resveratrol+benzo (a) pyrene group ( P < 0.001) . Conclusion:Resveratrol can inhibit the environmental pollutant benzo (a) pyrene-induced expression of inflammatory factor IL-1α in SZ95 sebocytes, which is likely mediated by the AhR and p38MAPK pathways.

Dengue fever is one of the most important vector-borne human diseases caused by mosquito vector Aedes aegypti and Aedes albopictus. Dengue virus can cause dengue fever, dengue hemorrhagic fever and dengue shock syndrome. There are no approved drugs for the treatment of dengue disease so far. In this paper, combined with the latest progress in the research of anti-dengue virus, the new progress in the research of anti-dengue virus drugs was reviewed from the aspects of protease inhibitors, virus polymerase inhibitors, entry inhibitors, virus replication-related host factor inhibitors, capsid protein inhibitors and nucleic acid inhibitors.

Objective To evaluate the effect of isotretinoin on expression of ache-associated inflammatory genes induced by peptidoglycan in human SZ95 sebocytes,and to explore the molecular mechanism underlying the treatment of acne with isotretinoin.Methods Cultured SZ95 sebocytes were divided into 3 groups:control group receiving no treatment,peptidoglycan group treated with 20 mg/L peptidoglycan alone,and costimulation group treated with 20 mg/L peptidoglycan combined with 10-5 mol/L isotretinoin.After 3-hour treatment,real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of pro-inflammatory cytokines including interleukin (IL)-1α,IL-1β,IL-6,IL-8,tumor necrosis factor (TNF)-α,Toll-like receptor 2 (TLR2) and MyD88 (a downstream gene of TLR2) in SZ95 sebocytes in the above groups.After 24-hour treatment,enzyme-linked immunosorbent assay (ELISA) was performed to detect levels of IL-1α,IL-1β,IL-6,IL-8 and TNF-α in the cell culture supernatant in the above groups.After 48-hour treatment,Western blot analysis was conducted to determine the protein expression of TLR2 and MyD88.Statistical analysis was carried out with SPSS 23 software by one-way analysis of variance (ANOVA) for the comparison among the 3 groups,and by Bonferroni method for multiple comparisons.Results The mRNA and protein expression of pro-inflammatory cytokines including IL-1α,IL-1β,IL-6,IL-8 and TNF-α all significantly differed among the 3 groups (all P ＜ 0.01),and was significantly higher in the peptidoglycan group than in the control group and costimulation group (both P ＜ 0.016 7).The mRNA expression of MyD88 also significantly differed among the control group,peptidoglycan group and costimulation group (6.707 ± 0.950,10.270 ± 0.477,7.892 ± 0.900 respectively,F =10.17,P ＜ 0.01),and was significantly higher in the peptidoglycan group than in the control group and costimulation group (t =4.740,3.298 respectively,both P ＜ 0.016 7).The mRNA and protein expression of TLR2 were markedly higher in the peptidoglycan group than in the control group,but did not differ between the peptidoglycan group and the costimulation group.Conclusion Isotretinoin can inhibit peptidoglycan-induced expression of inflammatory factors possibly associated with the occurrence of acne in human SZ95 sebocytes,likely by inhibiting the expression of MyD88,but not TLR2,in the innate immune response,which may be one of the mechanisms underlying the treatment of acne with isotretinoin.

Background: The coexistence of chronic obstructive pulmonary disease (COPD) and obstructive sleep apnea (OSA) is termed overlap syndrome (OS). COPD and OSA both have increased risks of developing cardiovascular diseases. This study aimed to explore if patients with OS exhibited a higher prevalence of cardiovascular complications, and if patients with OS exhibited vascular endothelial dysfunction and abnormalities in the cellular immune function of T lymphocytes. Methods: Totally 25 patients with stable COPD (COPD group), 25 patients with OSA (OSA group), 25 patients with OS (OS group), and 20 healthy adults (control group) were enrolled between January 2017 and December 2017 from the Respiratory Department of Tianjin Medical University General Hospital. The clinical characteristics of the four groups were collected and the expression levels of soluble vascular cell adhesion molecule-1 (sVCAM-1), tumor necrosis factor-α (TNF-α), and T-lymphocyte subsets were detected. One-way analysis of variance, χ² test and Pearson correlation were used to manage the data. Results: The prevalence of hypertension and coronary heart disease was significantly higher in the OS group than in the control, OSA, and COPD groups (χ² = 20.69, P < 0.05 and χ² = 11.03, P < 0.05, respectively). The levels of sVCAM-1 and TNF-α were significantly higher in the OS group than in other groups (F = 127.40, P < 0.05 and F = 846.77, P < 0.05, respectively). The percentage of CD4+ lymphocytes and CD4+/CD8+ were both significantly lower in the OS group than in any other group (F = 25.40, P < 0.05 and F = 75.08, P < 0.05, respectively). There were significantly negative correlations in the levels of sVCAM-1 and TNF-α with CD4+/CD8+ lymphocytes (r = –0.77, P < 0.05 and r = –0.83, P < 0.05, respectively). Conclusions The prevalence of hypertension and coronary heart disease was higher in patients with OS than in patients with either OSA or COPD alone. Patients with OS exhibited more severe vascular endothelial injury, stronger inflammatory response, and lower cellular immune function.

OBJECTIVE: To evaluate the clinical efficacy of warm moxibustion therapy in the recovery of quadriceps muscle strength in patients undergoing total knee arthroplasty (TKA) with analgesia of the femoral nerve block (FNB). METHODS: A total of 174 patients with KOA were randomized into a warm moxibustion group and a rehabilitation group, 87 cases in each group. In the warm moxibustion group, warm moxibustion combined with conventional quadriceps strength training were used. In the rehabilitation group, conventional quadriceps strength training was given. The warm moxibustion was applied at Liangqiu (ST 34) and Zusanli (ST 36), the treatment was given twice a day, 7 days for one course, with a total of 2 courses.The quadriceps muscle strength of the two groups was recorded and compared at 24 h before FNB, 24, 48, 72 and 96 h after surgery, and the resting and exercise VAS pain scores were also recorded at the same time point. And the first time for standing up and the first straight raising time in the two groups were compared, and the occurrence of adverse reactions in the two groups were observed. RESULTS: At 24, 48, 72 and 96 h after FNB, the quadriceps muscle strength in the warm moxibustion group was better than that in the rehabilitation group (<0.05, <0.01). At 72 h and 96 h after FNB, the resting and exercise VAS scores of the warm moxibustion group were lower than those of the rehabilitation group (both <0.001). The average first straight leg raising time in the warm moxibustion group was postoperative (31.03±10.78) h, and the time in the rehabilitation group was postoperative (47.23±15.78) h. The difference was statistically significant (<0.001). The average time of the first time for standing up in the warm moxibustion group was postoperative (25.76±7.00) h, and postoperative (33.12±11.18) h in the rehabilitation group. The difference was also statistically significant (<0.001). No adverse reactions occurred in both groups. CONCLUSION Warm moxibustion combined with conventional quadriceps strength training can improve the symptoms of quadriceps weakness in patients with femoral nerve block after total knee arthroplasty, and accelerate the recovery of joint function, which is superior to conventional quadriceps strength training.
Arthroplasty, Replacement, Knee ; Femoral Nerve ; Humans ; Moxibustion ; Muscle Strength ; Nerve Block ; Pain, Postoperative ; Quadriceps Muscle ; Treatment Outcome