Objective To construct mice hepatocellular carcinoma models with different tumor immune microenvironments(TIME)and explore the differences.Methods H22 and hepa1-6 were used to construct subcutaneous transplantation tumor model of C57 mice as homologous hepatocellular carcinoma cell lines(denoted as H22 group and hepal-6 group,each n=8),and the differences of TIME were evaluated.Immunohistochemistry was used to detect and quantify the infiltration of T cells,CD4+T cells,CD8+T cells,regulatory T cells and B cells in TIME.Flow cytometry was performed to detect the differences of composition of immune cell subpopulations in peripheral blood and tumor parenchyma.Gene expression profile characteristics of tumor tissue were analyzed based on high-throughput transcriptome sequencing technology,and enrichment analyses of immune-related signaling pathways were evaluated combined with gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG).Results H22 group showed cold and hepa1-6 group showed hot TIME characteristics.The number of T cells,CD4+T cells and CD8+T cells in tumor tissue of H22 group were all lower,while the proportion of T cells,CD4+T cells and CD8+T cells in peripheral blood were all higher than those of hepa1-6 group(all P＜0.05).Compared with H22 group,up-regulated genes of tumor tissue in hepa1-6 group expressed significantly enriched in tumor immune activation-related signaling pathways.Conclusion H22 and hepa1-6 hepatocellular carcinoma models showed distinct TIME characteristics of cold and hot tumors,respectively,and the amount of immune cells in tumor tissue of the former were significantly lower than those in the latter.

Objective To construct mice hepatocellular carcinoma models with different tumor immune microenvironments(TIME)and explore the differences.Methods H22 and hepa1-6 were used to construct subcutaneous transplantation tumor model of C57 mice as homologous hepatocellular carcinoma cell lines(denoted as H22 group and hepal-6 group,each n=8),and the differences of TIME were evaluated.Immunohistochemistry was used to detect and quantify the infiltration of T cells,CD4+T cells,CD8+T cells,regulatory T cells and B cells in TIME.Flow cytometry was performed to detect the differences of composition of immune cell subpopulations in peripheral blood and tumor parenchyma.Gene expression profile characteristics of tumor tissue were analyzed based on high-throughput transcriptome sequencing technology,and enrichment analyses of immune-related signaling pathways were evaluated combined with gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG).Results H22 group showed cold and hepa1-6 group showed hot TIME characteristics.The number of T cells,CD4+T cells and CD8+T cells in tumor tissue of H22 group were all lower,while the proportion of T cells,CD4+T cells and CD8+T cells in peripheral blood were all higher than those of hepa1-6 group(all P＜0.05).Compared with H22 group,up-regulated genes of tumor tissue in hepa1-6 group expressed significantly enriched in tumor immune activation-related signaling pathways.Conclusion H22 and hepa1-6 hepatocellular carcinoma models showed distinct TIME characteristics of cold and hot tumors,respectively,and the amount of immune cells in tumor tissue of the former were significantly lower than those in the latter.

Objective: To evaluate the effect of shear bond strength between resin cement and zirconia using SiO₂-ZrO₂ slurry coating. Methods: One hundred and forty pre-sintered zirconia discs were randomly divided into seven groups (n=20) according to the surface treatments: AS (as-sintered), SB (sand blasting with Al₂O₃), 2SiO₂-1ZrO₂ (2∶1 mole ratio SiO₂-ZrO₂ coating), 1SiO₂-1ZrO₂ (mole ratio 1∶1 SiO₂-ZrO₂ coating), 1SiO₂-2ZrO₂ (mole ratio 1∶2 SiO₂-ZrO₂ coating), 1SiO₂-3ZrO₂ (mole ratio 1∶3 SiO₂-ZrO₂ coating), 1SiO₂-4ZrO₂ (mole ratio 1∶4 SiO₂-ZrO₂ coating). Each zirconia disc was bonded to composite resin cylinder using resin cement. All specimens were stored in distilled water (37 ℃, 24 h). Each group was divided into two subgroups in which half specimens were tested using universal testing machine and another half specimens accepted artificial aging of 5 000 times thermocycling then tested. Scaning electron microscopy (SEM) was used to observe the micro-morphology of coating surface etched by hydrofluoric acid,then the coating thickness was measured. Results: Before artificial aging, 1SiO₂-1ZrO₂ showed a higher shear bond strength [(41.69±6.28) MPa] than all the other group (P<0.05). 1SiO₂-2ZrO₂ gained a higher strength than AS, SB, 1SiO₂-3ZrO₂ and 1SiO₂-4ZrO₂ (P<0.05). However, 1SiO₂-2ZrO₂ did not get a significant higher shear bond strength than 2SiO₂-1ZrO₂ (P>0.05). No significant differences were found among SB, 2SiO₂-1ZrO₂ and 1SiO₂-3ZrO₂ (P>0.05). After artificial aging, shear bond strength of all groups were decreased significantly besides 2SiO₂-1ZrO₂. 2SiO₂-1ZrO₂, 1SiO₂-1ZrO₂ and 1SiO₂-2ZrO₂ [(24.13±5.50), (22.28±4.40), (23.11±4.80) MPa] showed higher shear bond strength than SB and 1SiO₂-3ZrO₂ (P<0.05),no intergroup differences were observed (P>0.05). Shear bond strength of AS and 1SiO₂-4ZrO₂ fell to 0 MPa approximately. The SEM images of etched coating surface showed contraction fissure due to different thermal expansion coefficient between SiO₂ and ZrO₂ and intercrystal pores of zirconia. The thickness of coating was measured to be less than 30 μm. Conclusions: Mole ratio 1∶1 SiO₂-1ZrO₂ slurry coating showed the highest shear bond strength of resin cement to zirconia.
Dental Bonding ; Materials Testing ; Resin Cements/chemistry* ; Silicon Dioxide ; Surface Properties ; Zirconium

Gastrectomy ; Humans ; Lymph Node Excision ; Lymph Nodes ; Lymphatic Metastasis ; Retrospective Studies ; Stomach Neoplasms/surgery*

ObjectiveTo investigate the role of the serum inhibin B (INHB) level in evaluating the testicular function of the prepubertal patient with varicocele (VC) after high ligation of the spermatic vein (HLSV).

METHODSThis study included 31 prepubertal male patients with left VC, averaging 12.55 years of age and 9 complicated by right VC. We collected peripheral blood samples before and at 4, 12 and 26 weeks after HLSV as well as spermatic venous blood samples intraoperatively for determination of the levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), anti-sperm antibody (AsAb) and serum INHB by ELISA.

RESULTSCompared with the baseline, statistically significant differences were observed in the INHB level in the peripheral blood at 12 and 26 weeks after operation (［255.18 ± 69.97］ vs ［141.78 ± 59.82］ pg/ml, P < 0.05) and that in the spermatic venous blood intraoperatively (［255.18 ± 69.97］ vs ［412.44 ± 259.42］ pg/ml, P < 0.01). Spearman's analysis showed a negative correlation between the level of INHB and that of FSH (r = －0.224, P < 0.01).

CONCLUSIONSThe level of serum INHB in the peripheral blood of the prepubertal VC patient is decreased within 6 months after HLSV and negatively correlated with that of FSH. The levels of INHB and FSH may well reflect the testicular function of the prepubertal VC patient.

Adolescent ; Antibodies ; blood ; Biomarkers ; blood ; Child ; Follicle Stimulating Hormone ; blood ; Humans ; Inhibins ; blood ; Luteinizing Hormone ; blood ; Male ; Spermatozoa ; immunology ; Testosterone ; blood ; Varicocele ; blood

OBJECTIVETo observe effects of removing arms and ovarian on lumbar intervertebral disc and vertebral bone mineral density (BMD) by establishing rat model of lumbar intervetebral disc degeneration (IDD) with kidney deficiency, and to explore internal mechanism of disc degeneration, relationship between disc degeneration and osteoporosis.

METHODSThirty Sprague-Dawley female rats aged one month were randomly divided into control group, lumbar IDD group and lumbar IDD with kidney deficiency group (combined group), 10 rats in each group. Lumbar IDD group removed double arms, lumbar IDD with kidney deficiency group removed double arms after 3 months, both ovaries were removed. Vertebral bone mineral density were observed by Micro-CT scan; morphological changes were tested by safranine O-fast green staining; II, X collagen protein expression in the intervertebral disc were obsevered by immunohistochemistry; extracellular matrix gene expression were obsevered by real-time polymerase chain reaction (RT-PCR), in order to evaluate the effects of removed of forelimbs and double ovarian on degeneration and vertebral bone mineral density of intervertebral disc.

RESULTSMicro-CT scan showed osteoporosis in kidney deficiency group was obviously worse than other two groups; safranine O-fast green staining showed that intervertebral space became narrowed, intervertebral disc tissue degenerated obviously, chondral palte was underdeveloped in kidney deficiency group; immunohistochemistry showed that X collagen expression increased, type II collagen expression decreased in kidney deficiency group; RT-PCR showed that type II collagen expression in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P=0.000); Age 1 in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P= 0.000); while type X collagen expression was higher than control group, but no significant meaning; MMP-13 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P= 0.000, P=0.000); aggrecanase-2 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P=0.006, P=0.008).

CONCLUSIONRats model of lumbar disc degeneration established by removed forelimbs and ovariectomized can occure "bone like"--osteoporosis, which is similar with clinical kidney lumbar disc degeneration in tissue morphology, molecular cell biology expression.

Animals ; Collagen ; genetics ; metabolism ; Extracellular Matrix ; genetics ; metabolism ; Female ; Humans ; Intervertebral Disc Degeneration ; etiology ; metabolism ; physiopathology ; surgery ; Kidney ; physiopathology ; Osteoporosis ; complications ; genetics ; metabolism ; Ovariectomy ; adverse effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo compare the enhanced recovery program after surgery (ERAS) with conventional perioperative management in patients undergoing radical resection for colorectal cancer.

METHODSThe ERAS protocol included a combination of evidence-based and consensus methodology. A total of 597 consecutive patients undergoing elective colorectal resection were randomized to either the ERAS(n=299) or the control group(n=298). Outcomes related to nutrition and metabolism index, stress index, and recovery index were measured and recorded.

RESULTSDemographics and operative parameters were similar between the two groups(P>0.05). The nutritional status of patients in the ERAS group was improved after surgery compared with that of the control group. On postoperative day (POD) 1, the HOMA-IR in the ERAS group was significantly lower than that in the control group(P<0.01). The cortisol level in the control group was elevated on both POD 1(P<0.01) and POD 5(P<0.01) compared to the preoperative level. However, the cortisol level was not increased until POD 5(P<0.01) in the ERAS group. The levels of TNF-α, IL-1β, IL-6, and IFN-γ were reduced in the ERAS group, indicating less postoperative stress responses compared with the control group. In addition, ERAS group was associated with accelerated recovery of gastrointestinal function. The postoperative length of stay [(5.7±1.6) d vs. (6.6±2.4) d, P<0.01] and expense[(15 998±2655) RMB vs. (17 763±3059) RMB, P<0.01] were reduced in the ERAS group. Twenty-eight patients(9.4%) in the control group and 29(9.7%) in the ERAS group developed complications, while the difference was not statistically significant(P>0.05).

CONCLUSIONERAS protocol alleviates surgical stress response and accelerates postoperative recovery without compromising patient safety.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; surgery ; Female ; Humans ; Male ; Middle Aged ; Perioperative Care ; methods ; Prospective Studies ; Young Adult

Contributions of Xin'an medical school and physicians to acupuncture theory were introduced in the article. Academic theories or characteristics of several physicians of Xin'an school such as YANG Xuan-cao, WU Kun, WANG Ji, WU Yi-ding, ZHENG Mei-jian and XU Chun-fu, et al were sorted out. Contributions of inheriting and illustrations on acupuncture theory were analyzed so as to expound its significance and value on modern acupucture clinic.
Acupuncture ; education ; history ; manpower ; Acupuncture Therapy ; history ; China ; History, Ancient ; Humans ; Physicians ; history ; Schools, Medical ; history ; manpower

OBJECTIVETo examine whether the polymorphisms of endothelial nitric oxide synthase (eNOS) gene are associated with the susceptibility to high altitude pulmonary edema (HAPE) in Chinese railway construction workers at Qinghai-Tibet where the altitude is over 4 500 m above sea level.

METHODSA case-control study was conducted including 149 HAPE patients in the construction workers and 160 healthy controls randomly recruited from their co-workers, matching the patients in ethnicity, age, sex, lifestyle, and working conditions. Three polymorphisms of eNOS gene, T-786C in promoter, 894G/T in exon 7, and 27bp variable number tandem repeat (VNTR) in intron 4, were genotyped using polymerase chain reaction (PCR) and confirmed with DNA sequencing.

RESULTSThe frequencies of 894T allele and heterozygous G/T of the 894G/T variant were significantly higher in HAPE patients group than in the control group (P=0.0028 and P=0.0047, respectively). However, the frequencies of the T-786C in promoter and the 27bp VNTR in intron 4 were not significantly different between the two groups. Haplotypic analysis revealed that the frequencies of two haplotypes (H3,T-T-b, b indicates 5 repeats of 27 bp VNTR; H6, C-G-a, a indicates 4 repeats of 27 bp VNTR) were significantly higher in HAPE patients (both Pü0.0001). On the contrary, the frequencies of H1 (T-G-b) and H2 (T-G-a) were lower in HAPE patients than in healthy controls (both Pü0.001).

CONCLUSIONSTwo haplotypes (T-T-b and C-G-a) may be strongly associated with susceptibility to HAPE. Compared with the individual alleles of eNOS gene, the interaction of multiple genetic markers within a haplotype may be a major determinant for the susceptibility to HAPE.

Adolescent ; Adult ; Altitude ; Base Sequence ; Case-Control Studies ; DNA Primers ; Genotype ; Haplotypes ; Humans ; Male ; Middle Aged ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type III ; genetics ; Occupational Diseases ; enzymology ; genetics ; Polymorphism, Genetic ; Pulmonary Edema ; enzymology ; genetics ; Tibet ; Young Adult

Objective To investigate the epidemiological condition of the sub-health status of the young and middle-aged persons in urban and rural areas of Hubei province, and to explore the relationships among personality, quality of sleep and sub-health status. Methods Purposive sampling method and Personal Health Status Questionnaire developed by Chinese Academy of Medical Sciences were used to investigate the sub-healthy status of 3187 subjects aged 18-55 and evaluate the relationships among sub-health status, personality and the quality of sleep with structural equation model. Results Gender, education, marriage, age and occupation were factors influencing the status of sub-health. Among all of the subjects, sub-health status of females were more serious than that of males (P＜0.0001 of all). The divorced, separated and widowed ones were more likely to experience sub-health than other status of marriage (P= 0.0006, P=0.0102). The sub-health status of the subjects aged 30-55 in the urban (P=0.0197)and low-literacy residents in rural (P=0.0003)were among the worst. The condition of the sub-health in migrant workers and professional were worse compared to other occupational groups (P=0.0043). Structural equation model analysis revealed that sub-health status associated with the quality of sleep and the type of characteristics (P＜0.01 ). Conclusion Sub-health status did exist among the young and middle-aged persons and was influenced by the quality of sleep and personality. However, it could be adjusted by improving the quality of sleep and personality to prevent sub-health.