AIM:To investigate the effects of insulin on drug sensitivity of Etoposide(Vp-16) to MBA-MD-543(human breast cancer cell line) in vitro.METHODS:Insulin was applied directly to the MBA-MD-543 cultured in vitro.The effects of insulin and Vp-16 on the number of viable cell,the total number of cells were assayed by MTT method and cell count.RESULTS: Insulin could induce the cell growth and promote the cell metabolism at the concentration(4.0)-(32.0)(mU?ml~(-1))(8-18 hours).At the condition of same density of cells,the administration of insulin((7.5)(mU?ml~(-1))) before adding Vp-16 in 9-15 h,enhanced the chemocytotoxity of Vp-16((70.09)(?g?ml~(-1))) on human breast cancer cells as indicated by MTT colorimetry.CONCLUSION: Proliferation of human breast cancer cell line can be induced by insulin,and insulin can sensitize MBA-MD-543 to the anticancer activity of Vp-16 in vitro. The key of improving the chemotherapeutic effect is the selection of revulsant occasion and concentration.It is possible to increase the growth and metabolism of cancer cells first so as to enhance the chemosensibility,and then administer chemotherapeutic agents,thus improving their theraeutic effects.

Objective To investigate the impact of covariates on diagnostic test and assess the correlation between re-peated measurement data,this paper explores innovative modeling techniques of ROC curve.Methods We introduce the new ROC curve method based on generalized linear mixed effects model and apply Bayesian techniques to parameters estimation with Winbugs Software.Further,areas under the ROC curve(AUC)with different values of covariates could be calculated in terms of assessment.Results Cases analysis results indicate the proposed method efficiently explores the repeated measurement data and provides parameters with practical significance,serving as a golden reference.Conclusion The ROC curve based on generalized linear mixed effects models can be effectively used to solve the test accuracy evaluation problem of the repeated diagnostic trials.

To compare the performances of community family physicians in different patterns of chronic disease management.From June 2011 to April 2012,3 different models (A,B,C) of chronic disease management were employed for a total of 4972 patients.Statistics analyses were performed.The number of participants,session and average effective working time were different.In terms of standard management of hypertension,diabetic management rate and non on-site management rate,model C was superior to models A and B (P ＜ 0.01).And model A had the lowest rate of non on-site management (P ＜0.05).Though with each own advantage,three models are complementary.But model C reflects the residents' self-management concept of chronic disease.

Objective To explore the influence of dexmedetomidine combined with sufentanil on postoperative analgesia effect and immune function for patients undergoing lower limb fracture surgery. Methods One hundred cases patients of lower limb fracture undergoing open reduction and internal fixation of elective surgery in our hospital from January 2016 to November 2016 were selected ( ASAⅠ-Ⅱgrade, male 51 cases and female 49 cases, 30-65 years old, 50-75 kg). All cases were randomly divided into sufentanil group (Group S) and dexmedetomidine combined with sufentanil group (Group DS) according to the random number table, who used patient-controlled intravenous analgesia (PCIA). The PCIA drug formulations of two groups were as followings: sufentanil group (Group S) used 3 μg/h sufentanil+ondansetron 16 mg, dissolved in 0.9%sodium chloride injection 100 ml;dexmedetomidine combined with sufentanil group (Group DS) used dexmedetomidine 0.1μg/(kg · h)+sufentanil 2μg/h+ondansetron 16 mg, dissolved in 0.9%sodium chloride injection 100 ml;background infusion rate was 2 ml/h, and loading dose was 2 ml. Dose for patient-controlled analgesia (PCA) was 0.5 ml, and locking time was 15 min. The changes of pain, MAP, HR , sedation scores were recorded at 4 h (T1), 8 h (T2), 24 h (T3) and 48 h (T4) after operation; nausea and vomiting, hypotension, bradycardia, respiratory depression and other adverse reactions were aslo recorded at the same time. At 10 min before induction of anesthesia (T0) and T1-T4 after operation, the CD3+, CD4+, CD8+and NK cell activity of peripheral blood T lymphocyte subsets were determined using flow cytometry, and CD4 +/CD8 + ratio was calculated. Results The levels of MAP and HR in DS group at each time point after operation were lower than those in group S (P<0.05), the level of visual analogue score (VAS) in DS group at different time points were significantly lower than those of s group: (1.8 ± 0.3) scores vs. (2.5 ± 0.5) scores, (1.1 ± 0.5) scores vs. (1.9 ± 0.3) scores, (1.0 ± 0.5) scores vs. (1.8 ± 0.5) scores, (0.8 ± 0.3) scores vs. (1.5 ± 0.6) scores (P < 0.05), Ramsay Sedation score was significantly higher than that of s group: (3.5 ± 0.3) scores vs. (2.4 ± 0.6) scores, (3.3 ± 0.5) scores vs. (2.5 ± 0.3) scores, (3.5 ± 0.6) scores vs. (2.3 ± 0.5) scores, (3.2 ± 0.4) scores vs. (2.2 ± 0.5)scores (P<0.05);nausea and vomiting occurred in two patients after the operation of group DS , which was significantly lower than that of S group (P<0.05);compared with those at T0, CD3+, CD4+, CD4+/CD8+and NK cells in the two groups of patients decreased significantly at the time of T1-T4 (P<0.05);The levels of CD3+, CD4+, CD4+/CD8+and NK cells were significantly higher in group DS at T1- T4 than those in group S(P < 0.05). Conclusions Dexmedetomidine combined with sufentanil for analgesia in patients with lower limb fracture surgery has better effect. It decreases the incidence of nausea and vomiting, and improves the cellular immune function of patients.

BACKGROUNDAltered immunoresponse is associated with tumorigenesis and cancer progression. This study assessed the levels of tumor-infiltrating CD3 + or CD8 + T lymphocytes and interleukin-2 (IL-2) protein in radically resected non-small cell lung cancer (NSCLC) tissues to predict overall survival (OS) of the patients.

METHODSParaffin-embedded tissue specimens from 129 NSCLC patients were retrospectively collected for immunostaining of CD8 + , CD3 + , and IL-2 expression. Clinicopathological and survival data were collected and analyzed using the Chi-squared test, Kaplan-Meier curves, and the log-rank test or the Cox regression model.

RESULTSThe data showed a significant inverse association between CD8 + T lymphocyte levels and IL-2 expression (r = -0.927; P = 0.000) and between the levels of CD8 + and CD3 + T lymphocytes (r = -0.722; P = 0.000), but a positive association between CD3 + T lymphocyte levels and IL-2 expression (r = 0.781; P = 0.000) in NSCLC tissues. Furthermore, the levels of CD3 + and CD8 + T lymphocytes and IL-2 expression were associated with tumor stage (P = 0.023, 0.006, and 0.031, respectively) and the level of CD8 + T lymphocytes was associated with the patient gender (P = 0.024). In addition, the levels of CD8 + T lymphocytes were associated with an unfavorable 5-year OS, whereas patients with high levels of CD3 + T lymphocytes in tumor lesions and IL-2-expressing tumors had significantly better 5-year OS rates than patients with low levels.

CONCLUSIONSThe levels of CD8 + T cells in tumor lesions and IL-2 expression were both independent predictors of OS for these NSCLC patients. Thus, the detection of tumor-infiltrating CD3 + or CD8 + T lymphocytes and IL-2 expression could be useful to predict the prognosis of radically resected NSCLC patients.

CD3 Complex ; metabolism ; CD8-Positive T-Lymphocytes ; metabolism ; Female ; Humans ; Immunohistochemistry ; Interleukin-2 ; metabolism ; Lung Neoplasms ; immunology ; metabolism ; Lymphocytes, Tumor-Infiltrating ; metabolism ; Male ; Prognosis

The development of cervical cancer is associated with persistent infection of high-risk human papillomavirus. The detection and genotyping of HPV could be used to evaluate the epidemiology of HPV infections, monitor HPV vaccine efficacy, and screen cervical cancer. There are a lot of commercially available molecular tests for HPV, based on different methodologies and detection systems. In principle, it mainly includes two categories, namely signal amplification and target amplification. Most of them are based on PCR amplification, such as fluorescent PCR, PCR-reverse hybridization, etc. The performances of detection reagents are different. In addition, HPV genotyping assays based on next-generation sequencing and quantitative HPV detection kits are developed. However, only a small number of commercial assays have been clinically verified. A large number of assays which may bring greater values in the screening of cervical cancer are needed to be clinically validated.

BACKGROUND:Immunosuppression leads to impaired body immune function and aggravates the disease.Herb-partitioned moxibustion can effectively regulate immune function and improve immunity in the body,but its regulatory mechanism has not been elucidated. OBJECTIVE:To sequence immunosuppressed model rats treated with herb-partitioned moxibustion using bioinformatics techniques based on transcriptomics and to explore the mechanisms by which it regulates immunity. METHODS:Twenty-four Sprague-Dawley rats were randomly assigned to three groups:control,model,and herb-partitioned moxibustion groups,with eight rats in each group.The model and herb-partitioned moxibustion groups were subjected to establishment of an immune suppression model by intraperitoneal injection of cyclophosphamide at a dose of 35 mg/kg for 3 consecutive days.No interventions were administered to the control and model groups after modeling.In contrast,the herb-partitioned moxibustion group received moxibustion treatment at Zhongwan,Shenque,Guanyuan,and Zusanli acupoints using a combination of moxa and herbal cakes,once a day,for 10 consecutive days,with samples being collected the day after the end of the intervention.Peripheral blood was collected from all groups of rats to measure their white blood cell count.RNA-seq was performed on the Illumina sequencing platform,and differentially expressed genes were selected for bioinformatics analysis using the GO and KEGG databases. RESULTS AND CONCLUSION:Compared with the control group,the model group exhibited a significant decrease in white blood cell count(P<0.001).RNA-seq analysis identified 3 026 differentially expressed genes between the model and control groups,with 1 565 upregulated and 1 461 downregulated.There were 535 differentially expressed genes identified between the herb-partitioned moxibustion group and the model group,with 280 upregulated and 255 downregulated.The Venn diagram analysis revealed that 159 genes were downregulated in the model group compared with the control group.However,after moxibustion with herbal cakes,these genes were upregulated.Protein-protein interaction network analysis identified 10 core targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,Mx1,Syk,Hspa1a,and Ret.According to GO and KEGG analyses,moxibustion with herbal cakes regulated the body through pathways related to immune response,viruses,angiogenesis,and the autoimmune system.To conclude,there is a significant association between herbal cake-separated moxibustion intervention and immune suppression targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,and Mx1.The intervention exhibits regulatory effects in the pathways related to immune responses,viral activities,and angiogenesis.

Objective:To compare similarities and differences in expression changes of cytotoxic T-lymphocyte antigen-4(CT-LA-4)and programmed death receptor 1(PD-1)in immunosuppressed rabbits under different moxibustion interventions.Methods:Twenty large-eared white rabbits were randomly divided into normal group,model group,moxa stick moxibustion(MSM)group and herbal cake-partitioned moxibustion(HPM)group,with five rabbits in each group.CTX-induced immunosuppressed models were prepared by intraperitoneal injection for 7 consecutive days.After successful modeling,MSM and HPM were performed on alternate days for 10 treatments.Rabbits were anesthetized after treatment,and serum,liver and spleen were collected.Serum PD-1 and PD-L1 contents were detected by ELISA.PD-1 in liver tissue was detected by immunohistochemistry,and CTLA-4 mRNA in liver and spleen tissues were detected by RT-qPCR.Results:Both HPM and MSM could reduce PD-1 and PD-L1 levels due to immunosuppression,and could effectively suppress elevated levels of CTLA-4 in spleen and PD-1 and CTLA-4 in liver,which were statistically different compared with immunosuppression model group(P<0.05).Pearson correlation test showed that CTLA-4 and PD-1 in liver tissue had significant positive correlation(r=0.780 7,P<0.001).Conclusion:HPM can improve body immune function by regulating multiple immunosuppressive sites.

ObjectiveTo explore the mechanism of herbal cake-separated moxibustion using the classical formula Xiaoyaosan in alleviating neuroimmune inflammatory responses in chronic fatigue syndrome （CFS） rats， based on the regulation of the toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear transcription factor-κB （NF-κB） signaling pathway. MethodsFifty SPF-grade SD rats aged 6-8 weeks were randomly divided into five groups： Normal group， model group， sham herbal cake moxibustion group， Chinese medicine intragastric administration group， and herbal cake-separated moxibustion group， with 10 rats in each group. Except for the normal group， all other groups underwent a 21-day modeling process， followed by behavioral testing. The herbal cake-separated moxibustion and sham herbal cake moxibustion groups received interventions at the Shenque （CV8）， Guanyuan （CV4）， Zusanli （ST36）， and Qimen （LR14） acupoints. The Chinese medicine intragastric administration group was treated with a Xiaoyaosan suspension via gavage. Behavioral tests were conducted after 10 days of continuous intervention. Serum levels of interleukin （IL）-1β， IL-6， and tumor necrosis factor-α （TNF-α）， as well as hippocampal levels of IL-1β， IL-6， TNF-α， and NF-κB， were detected by enzyme-linked immunosorbent assay （ELISA）. Morphological changes in the hippocampus were observed using hematoxylin-eosin （HE） staining. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect mRNA expression levels of TLR4， MyD88， and NF-κB in the hippocampus. Western blot analysis was performed to detect the relative expression levels of TLR4， MyD88， NF-κB， and p65 proteins in the hippocampus. ResultsCompared with the normal group， the model group showed a significant decrease in upright times during the open field test （P<0.01）， as well as significant reductions in total movement distance， resting time， and center region duration （P<0.01）. In the tail suspension test， immobility time increased （P<0.01）， and struggle times decreased （P<0.01）. Serum and hippocampal levels of IL-1β， IL-6， and TNF-α， as well as hippocampal NF-κB levels and TLR4， MyD88， and NF-κB mRNA expression， were significantly elevated （P<0.01）. After treatment， compared with the model group， the total movement distance and upright times in the open field test were significantly increased in all treatment groups （P<0.01）， while resting time and center region duration were notably prolonged （P<0.05， P<0.01）. Immobility time in the tail suspension test was significantly shortened （P<0.01）， and struggle times significantly increased （P<0.05）. Serum and hippocampal levels of IL-1β， IL-6， TNF-α， hippocampal NF-κB levels， and TLR4 and NF-κB mRNA expression were significantly reduced （P<0.05， P<0.01）. Compared with the sham herbal cake moxibustion group， the herbal cake-separated moxibustion group showed a significant extension in center region duration during the open field test （P<0.05） and a significant increase in upright times （P<0.01）. In the tail suspension test， immobility time was reduced （P<0.01）， and struggle times increased （P<0.01）. Serum TNF-α levels in the Chinese medicine intragastric administration group were significantly reduced （P<0.01）， while serum IL-6 levels， as well as hippocampal levels of IL-1β， TNF-α， NF-κB， and TLR4， MyD88， and NF-κB mRNA expression， were significantly decreased in both the Chinese medicine intragastric administration group and the herbal cake-separated moxibustion group （P<0.05， P<0.01）. Compared with the Chinese medicine intragastric administration group， the herbal cake-separated moxibustion group exhibited significantly increased upright times in the open field test （P<0.01）. In the tail suspension test， immobility time was reduced （P<0.01）， and struggle times increased （P<0.01）. Serum IL-1β， hippocampal TNF-α levels， and TLR4， MyD88， and NF-κB mRNA expression were significantly decreased （P<0.05， P<0.01）. ConclusionHerbal cake-separated moxibustion effectively improves fatigue and memory function in CFS rats， regulates neuroimmune inflammatory responses， and its mechanism may be related to the modulation of the TLR4/MyD88/NF-κB signaling pathway.

OBJECTIVETo compare the difference of serum sex hormone between female patients with post-adolescent acne and healthy women, and to explore the efficacy and action mechanism of acupoint catgut embedding, fire needle, auricular acupuncture on skin lesion in female patients of post-adolescent acne.

METHODSA total of 107 female patients of post-adolescent acne were divided into an integrated acupuncture group (54 cases, 4 cases were excluded) and a medication group (53 cases, 5 cases were excluded). The patients in the integrated acupuncture group were treated with comprehensive treatment of acupoint catgut embedding, fire needle, auricular acupuncture; the acupoint catgut embedding was applied at Dazhui (GV 14), Yintang (GV 29), Yangbai (GB 14) through Yuyao (EX-HN 4) and other acupoints based on syndrome differentiation; the fire needle was applied at skin lesion; the auricular acupuncture was applied at erjian (HX), e (AT), kou (CO), etc. The patients in the medication group were treated with oral administration of tanshinone capsules (4 capsules each time, 3 times a day) and external use of adapalene gel (one treatment per day at night). Patients in the two groups were treated for 8 weeks. The skin lesion of acne was evaluated before treatment as well as 4 weeks and 8 weeks after treatment in the two groups; the serum levels of testosterone (T) and estradiol (E) were tested 24 hours before menstruation in the integrated acupuncture group (50 cases) and healthy control group (46 cases), and the change of serum sex hormone after treatment was observed in 21 patients with sex hormone disorder in the integrated acupuncture group.

RESULTSBefore treatment, the level of E in the integrated acupuncture group was significantly lower than that in the healthy control group (<0.01), but T/E in the integrated acupuncture group was significantly higher than that in the healthy control group (<0.01). After treatment, the level of E was significantly increased (<0.01) and T/E was reduced (<0.01) in the 21 patients with sex hormone disorder in the integrated acupuncture group. The skin lesion scale of acne was significantly reduced in the two groups after 4-week and 8-week treatment (all <0.01); the difference between the two groups was not significant after 4-week treatment (>0.05); the skin lesion scale of acne in the integrated acupuncture group was significantly lower than that in the medication group after 8-week treatment (<0.01). The efficacy between the two groups was not significant after 4-week the treatment (>0.05); after 8-week treatment, the cured and effective rate was 66.0% (33/50) in the integrated acupuncture group, which was superior to 45.8% (22/48) in the medication group (<0.05).

CONCLUSIONCompared with healthy women, the level of serum sex hormone of E is reduced in the female patients of post-adolescent acne, resulting in relative increased level of T; the acupoint catgut embedding, fire needle, auricular acupuncture have better efficacy than medication for post-adolescent acne, which have regulation effects on sex hormone disorder.