Objective To investigate the expression of CD86 on systemic lupus erythematosus (SLE) peripheral B cells and the effect of triptolide (TL) on it.Methods The percentage of CD86 + B cells was detected by flow cytometer when the peripheral blood mononuclear cells (PBMC) was freshly seperated or after cultured with TL in different concentration for 48 hours.Results The percentage of CD86 + B cells of SLE patients was higher than that of normal controls either when the PBMC was freshly separated ( P

To investigate the enhancement of immunological activity of CpG ODN by chitosan gene carrier in mice, the effect of lymphocyte proliferation was detected in mice by using MTT, the levels of IgG and cytokines (IL-2 and IL-12) in serum were measured by ELISA and peripheral blood T lymphocyte subsets CD4(+), CD8(+) were analyzed by flow cytometry. Our results showed that spleen lymphocytes isolated from the CS-CpG ODN group of mice showed the strongest proliferation (SI=1.551), and the levels of IgG, IL-2 and IL-12 in serum were higher than those of other groups. Compared with the immunization with CpG ODN, the immunization with CS-CpG ODN gene carrier was more efficient in up-regulating the percentage of CD4(+)T cells and the ratio of CD4(+)/CD8(+) of mice. It was concluded that CS gene carrier of CpG ODN was much more effective in improving immunity of CpG ODN in mice.

OBJECTIVE: To study the expression of nuclear factor kappa B p65 in hyperlipemia model of mice, and the relationship between hyperlipemia and deaf. METHOD: Twenty mice were divided into two group. The hyperlipemia diet group was established in ten mice,and the normal diet group was served as normal control. Six weeks later, immunohistostaining was used to detected the express of NF-kappa B p65 in all mouse cochlear. ABR threshold was obtained from both normal group and hyperlipemia group. RESULT: Immunoreactivity NF-kappa B p65 in mouse cochlea of hyperlipemia was localized in the organ of Corti, tectorial membrane, stria vascularis, spiral ligament, spiral ganglion and nerve fibers. The NF-kappa B p65 expression was markedly increased in mouse cochlea of hyperlipemia ABR threshold was significant difference between hyperlipemia group mice and control group mice (P < 0.01). CONCLUSION The expression of NF-kappa B p65 in mouse cochlea can be induced by hyperlipemia. And ABR threshold increased in hyperlipemia group mice. This shows that hyperlipidemia can damage acouesthesia of mice.
Animals ; Cochlea ; metabolism ; Disease Models, Animal ; Hyperlipidemias ; metabolism ; Mice ; Mice, Inbred Strains ; Transcription Factor RelA ; metabolism

To investigate the enhancement of immunological activity of CpG ODN by chitosan gene carrier in mice, the effect of lymphocyte proliferation was detected in mice by using MTT, the levels of IgG and cytokines (IL-2 and IL-12) in serum were measured by ELISA and peripheral blood T lymphocyte subsets CD4+, CD8+ were analyzed by flow cytometry. Our results showed that spleen lymphocytes isolated from the CS-CpG ODN group of mice showed the strongest proliferation (SI =1.551), and the levels of IgG, IL-2 and IL-12 in serum were higher than those of other groups. Compared with the immunization with CpG ODN, the immunization with CS-CpG ODN gene carrier was more efficient in up-regulating the percentage of CD4+ T cells and the ratio of CD4+/CD8+ of mice. It was concluded that CS gene carrier of CpG ODN was much more effective in improving immunity of CpG ODN in mice.

Objective In order to explore the role of autophagy in the development of systemic lupus erythematosus (SLE),we measured the expression of autophagy related gene microtubule-associated protein 1 light chain 3 (LC3),Atg5,Beclinl,Atg7 and the incidence of autophagy in T cells from patients with SLE.Methods The mRNA levels of LC3,Atg5,Beclinl,Atg7 in T cells from 67 SLE patients and 31 healthy individuals were detected by real-time quantitative polymerase chain reacton (qPCR) technique.Autophagy in T cells from 17 SLE patients and 11 healthy controls was also determined by flow cytometry (FACs).The correlation of Atg7 mRNA expression with clincal features was then analyzed.The differences between the two groups were tested by t-test and x2 tcst,all data were analyzed by statistical and service solutions (SPSS) 17.0 software.Results The mRNA levels of LC3 and Atg7 (ΔCT value) in SLE patients were obviously down-regulated as compared to healthy populations (P=0.010,P=0.002),paralleled with the decreased autophagy rate detected by flow cytometry in T cells of SLE patients [(3.7±1.9)％ vs (6.6±1.4)％,t=4.132,P=0.000].Also,the protein expression levels of LC3-Ⅱ in T cells of SLE patients (LC3-Ⅱ/GAPDH) was significantly lower than those in healthy controls (0.21±0.08 vs 0.34±0.11,t=1.846,P=0.047).Moreover,Atg7 mRNA expression levels were found to be negatively correlated to autophagy rate (r=-0.492,P=0.008).However,when comparing the clinical features of 24 SLE patients with decreased Atg7 mRNA expression (ΔCT value＞9.86) to 43 SLE patients with normal or high Atg7 mRNA expression (ΔCT value ＜9.86),increasing trend of incidence of arthritis,blood involvement and CNS was noted in patients with decreased Atg7 mRNA expression.However,there was a significant difference between the two groups in the incidence of renal involvement and anti-dsDNA antibody and SLEDAI (P=0.008,P=0.018,P=0.035).Conclusion The impaired autophagy resulted from down-regulated LC3 and Atg7 mRNA levels in T cells from SLE patients indicates that autophagy plays a role in mediating the occurrence and development of SLE,which might be through unable to clean harmful molecules effectively.

Objective To investigate the application valve of real-time fluorescence quantitative polymerase chain reaction(RT-PCR) for the detection of tumor M2-pyruvate kinase(tM2-PK) DNA in patients with colorectal cancer(CRC).Methods Fragment of tM2-PK DNA(162 bp) was amplified and inserted into PGM-T vector to construct recombinant plasmid,which was used to develop RT-PCR method.Sensitivity,specificity and repeatability of RT-PCR for the detection of tM2-PK were analyzed.From Jan.2014 to Jun.2016,200 CRC patients and 100 healthy subjects were enrolled and detected for fecal and serum tM2-PK DNA by using RT-PCR,and the detected results were compared with those detected by using enzyme linked immunosorbent assay(ELISA).Results Recombinant plasmid was successfully constructed,which was certified by sequencing.The sensitivity of RT-PCR for the detection of tM2-PK DNA was 10 copy/mL,with high specificity and 0.3%-2.9% of coefficient of variation.In patients,the positive rate of fecal tM2-PK DNA,detected by RT-PCR,was 92.50%,and that of ELISA to detect tM2-PK was 80.00%.Fecal and serum levels of tM2-PK were correlated with the pathologic stages of tumour.Conclusion Self-established RT-PCR could be specificity and sensitivity for the detection of fecal tM2-PK,which could be used for the early diagnosis of CRC.

Objective In order to explore the effect of 25-(OH)D3 on monocyte chemoattratant protein (MCP)-1 expression from patients with system lupus erythematosus (SLE),we detected the level of active vitamin D and the expression of MCP-1 mRNA in patients with SLE,and analyzed the correlation between them.Methods The level of serum 25-(OH)D3 and mRNA expression of MCP-1 in 154 SLE patients and 31 healthy individuals were detected by enzyme-linked immuno sorbent assay (ELISA) and real time quanti-tative pol ymerase chain reaction (PCR) respectively.We also analyzed the correlation between serum 25-(OH)D3 level and the expression of MCP-1 mRNA,then analyzed the function of 25(OH)D3 on the regula-tion of MCP-1 mRNA expression in vitro.The differences between the two groups were tested by t-test and x2 test,multiple data were tested by one-way ANOVA and the correlation was analyzed by Pearson's correlation,all data were analyzed by statistical product and service solutions (SPSS) 17.0 software.Results The serum 25(OH)D3 levels in SLE group (20±11) ng/ml was significantly lower than normal control group (29±11) ng/ml (t=4.198,P＜0.01),and the ratio of the serum levels of vitamin D deficiency in SLE group were significantly higher than that of normal control group [55.8％(86/154) vs 22.6％(7/31),x2=11.421,P=0.001].The expression level of MCP-1 mRNA in PBMCs from the normal control group was significantly lower than the SLE group (1.14±0.27 vs 1.44± 0.31,t=3.277,P=0.001),serum 25(OH)D3 level and MCP-1 mRNA expression in patients with SLE PBMCs were significantly negatively correlated (r=-0.289,P＜0.01).Further study found that 25-(OH)D3 inhibited MCP-1 mRNA expression in PBMCs from SLE patients depending on the concentration.Conclusion The decreased 25-(OH)D3 level and up-regulated MCP-1 mRNA expression suggestthat MCP-1 may play an important role in SLE pathological process.

Objcetive: To explore the treatment of long segment defect of tibia by using tensor fascia lata combined with iliac flap or deep circumflex iliac pedicle iliac flap. Methods: From February 2012 to August 2017, The People′s Hospital of Zun Yi City Bo Zhou District treated 16 patients who had long segment defect of tibia.There were 11 males and 5 females, age from 22 to 58 years old, the average age was 42 years old. Iliac flap grafting with tensor fascia lata combined with iliac flap or deep circumflex iliac pedicle was used to treat the defect of long segment of tibia. There were 4 cases with simple tibial defect and 12 cases with skin defect. The longest tibial defect was 5-8 cm. Results: In this study, four patients used iliac flaps with deep circumflex iliac pedicle, the area of flaps ranged from 2.5 cm×5.0 cm to 5.0 cm×10.0 cm, while the area of iliac flaps ranged from 5.0 cm×2.5 cm to 8.0 cm×4.0 cm. Twelve patients used grafting with tensor fascia lata combined with iliac flap, the area of flaps ranged from 5.0 cm×12.0 cm to 12.0 cm×23.0 cm, while the area of iliac flaps ranged from 7.0 cm×2.0 cm to 8.0 cm×4.0 cm. All 16 cases of bone flap were survived, fracture healing, without surgical complications. The average follow-up period was 1.5 years, the flaps had good appearance in 10 cases and was slightly bloated in 6 cases; the ankle had normal motion in 14 cases and had poor dorsal extension in 2 cases. X-ray films showed that the bone flap repaired the bone defects and reached bone healing. Conclusions Vascularized tensor fascia lata combined with iliac flap or deep circumflex iliac pedicle iliac flap grafts increase local blood supply and accelerate the process of fracture healing.

Objective:To investigate the primary tumor volume change and timing of radiotherapy for patients with stage Ⅳ non-small cell lung cancer with EGFR mutation during molecular targeted therapy.Methods:Simulated CT scanning measurement and analysis were performed to observe the volume changes of primary tumors before and after treatment with a time interval of 10 days in this prospective study. Positioning and volume measurement were terminated when the volume change was 5% or less between two time points before and after treatment or 90 days after treatment. Primary tumor radiation therapy was then performed, acute radiation-induced injury was recorded, and the implementation and simulation of related parameters of radiotherapy plans were compared.Results:Twenty-nine of 30 cases were included in the analysis (1 case dropped off). After EGFR-TKIs treatment, the volume of all primary tumors was decreased, but the shrinking rate was inconsistent with the speed. Until the last simulated CT scanning, the maximum and minimum shrinking rates were 90% and 28%, respectively. There was no case of termination within 30 days of treatment, and the average tumor volume was significantly decreased within 40 days and the average tumor volume significantly differed every 10 days ( P<0.001). After 40 days, the volume shrinking rate of primary tumors ≤5% gradually appeared, and one patient presented with a volume shrinking rate of >5% on 90 days. During this time, the average volume shrinking rate slowed down and became stable, ranging from 49.15% to 54.77%. Moreover, the average volume continued to gradually shrink after slight increase at 70 days. There was no significant difference in the average volume every 10 days ( P>0.05). After the termination of simulated CT scanning, the dose of primary tumor was (69±7) Gy for patients receiving radiotherapy. Two patients had grade 2 acute radiation-induced pneumonitis and 3 patients had grade 3 acute radiation-induced pneumonitis. In addition, 1 patient had grade 2 radiation-induced esophagitis. According to the technology and dose parameters of radiotherapy plan, simulated radiotherapy plans before and 40 days after EGFR-TKIs treatment were designed. The timing of implementation plan was significantly better than that before EGFR-TKIs treatment (all P<0.05), whereas it was similar to that at 40 days after EGFR-TKI treatment ( P>0.05). Conclusions:The primary tumor shrinking rate is gradually slowed down over time after EGFR-TKIs treatment in patients with stage Ⅳ non-small cell lung cancer. The average tumor volume is significantly decreased within 40 days and then the shrinking rate becomes slow. The tumor shrinking rate of each case is inconsistent. Radiotherapy at 40 days after treatment is probably the optimal timing to obtain high dose and control radiation-induced injury.

Objective:To analyze the mediastinal displacement of target volume in the postoperative radiotherapy (PORT) process for non-small cell lung cancer (NSCLC) and the value of mid-term evaluation.Methods:For 100 patients with postoperativeN 2 stage NSCLC, R 1-2 and any N staging, bone anatomy was utilized to measure the change of the first and second CT localization on the same level. Statistical analysis were performed using the WilCoxon, Kruskal-Wallis and χ2 tests. The cut-off values were calculated with the receiver operating characteristic (ROC) curve. Results:Among the included patients, in the PORT process, the mediastinal displacement in the x (front and rear), Y (left and right) and Z (upper and lower) directions were 0.04-0.53 cm, 0.00-0.84 cm and 0.00-1.27 cm, respectively, and the order of mediastinal displacement distance wasz > Y> X,respectively. According to the ROC curve calculation, the cut-off values were 0.263, 0.352 and 0.405, respectively, which were greater than the cut-off values in 25 cases (25%), 30 cases (30%) and 30 cases (30%), respectively. There was significant difference in the three-dimensionalmediastinal displacement ( P=0.007, <0.001 and<0.001). The mediastinal displacement in thex, Y and Z directions had no statistical significance regarding resection site ( P=0.355, 0.239 and 0.256) and operation mode ( P=0.241, 0.110 and 0.064). Comparative analysis of modified whole group mediastinal shift> and cut-off values, medium-simulation (m-S) and the originally planned radiotherapy shown that there was no significant difference in the incidence of radiation esophagitis (RE) and radiation pneumonitis in PORT patients (all P>0.05); however, the incidence of ≥grade 3 RE in the modified plan after m-S was significantly lower than that in the originally planned PORT patients, which were 0 and 7%, respectively ( P<0.001). Conclusions:Mediastinal displacement exists in the PORT process of N 2 or/and R 1-2 cases after radical operation of NSCLC, and obvious movement occurs in 20%-30% of patients. Relocating and modifying the target volume and radiotherapy plan in the middle of the PORT process is beneficial to quality assurance and quality control.