Objective To evaluate the enhancement of chemosensitivity of celecoxib, a specific cyclooxygenase-2 (COX-2) inhibitor, on leukemia HL-60 cell line in vitro, and explore the possible mechanisms. Methods MTT assay was used to assess the cytostatic efficacy of doxorubicin in the absent or present of different doses of celecoxib on HL-60 cell. The apoptosis of HL-60 cells was measured by flow cytometry (FCM). Gene expressions of Survivin was examined by reverse transcription-polymerase chain reaction (RT-PCR). Protein of survivin was detected by Western blotting. Results Celecoxib could increase the cytostatic efficacy of doxorubicin on HL-60 cells. HL-60 cells were treated with increasing doses of doxorubicin in absence or presence of celecoxib (5 μmol/L, 10 μmol/L), IC50 were 0.48 μg/ml, 0.25 μg/ml and 0.16 μg/ml, respectively. Doxorubicin combined with low dose of celecoxib could induce the down-regulation of mRNA and protein of Survivin. Apoptosis rate of HL-60 cells treated with both 0.10 μg/ml doxorubicin and celecoxib(5 μmol/L, 10 μmol/L) were (13.07±1.66) % and (22.36±1.84) %, respectively, while it was (5.72±1.25) % in HL-60 cells treated with 0.10 μg/ml doxorubicin alone, with significant difference (P＜0.01).Conclusion Celecoxib could enhance the chemosensitivity of doxorubicin on leukemia HL-60 cell, which involves in increasing the apoptosis of HL-60 cells by down-regulation expression of Survivin.

ObjectiveTo explore the effects and possible mechanisms of VCR combined with low dose cyclophosphamide(CTX) intermittently to treat severe systemic lupus erythematosus(SLE).It is assumed that this might be a new combination therapy for SLE and expected to improve the overall prognosis and outcome of SLE.MethodsMurine chronic graft-versus-host disease(cGVHD) model were developed for study.They were randomly divided into the control group,vincristine (VCR) pulse therapy group,CTX pulse therapy group,CTX every other day(EOD) group,VCR+CTX combination group.One way ANOVA and repeated measure variance analysis were used for statistical analysis.Results① Six weeks after cGVHD models were set up,the average 24-hour urine protein quantification was(5.02±0.88) mg,anti-dsDNA antibody was positive,and Ⅳ LN pathology could be observed histologically in the model murine.So cGVHD models were successfully developed.② Significantly difference in decreasing of 24-hour urine protein quantification was found in the CTX EOD group,VCR+CTX combination group and other groups (P＜0.01).Significant decrease in Cr,ALT,anti-dsDNA,was found in the CTX EOD group,VCR+CTX combination group,CTX pulse therapy group and other groups(P＜0.05).Decrease in urine MCP-1 and TGF-β1 could be detected,and statistical significant difference in these parameters could be found in the CTX EOD group,CTX pulse therapy group,VCR+CTX combination group and other groups (P＜0.01).MCP-1 and TGF-β1'expression in model kidney were reduced in the CTX EOD group,VCR+CTX combination group and had statistical significant difference in the CTX EOD group,VCR+CTX combination group,VCR pulse therapy group,and CTX pulse therapy group.③ VCRand CTX combination treatment was effectivein 24-hour urine protein quantification,blood Cr,ALT,anti-dsDNA and urine MCP-1,as well as urine TGF-β1 (P＜0.05).Conclusion ① The combination of VCR and CTX is synergistic in decreasing 24-hour urine protein quantification,Cr,and the expression of MCP-1,TGF-β1.② The adverse effect of VCR+CTX combination group is similar to VCR pulse therapy group and CTX pulse therapy group.

Objective To investigate the tumor inhibition effect of paclitaxel long-circulating thermo sensitive liposomes (PLTL) in mice bearing Lewis lung carcinoma cells. Methods A tumor-bearing mouse model was established, and the mice were randomly divided into five groups: control, heating, paclitaxel (PTX), paclitaxel thermo sensitive liposomes (PTL), and PLTL groups. The living status was observed in the mice. The volume and weight of the tumor were measured. The morphological changes in the tumor cells were observed f by HE staining and apoptosis of the tumor cells was determined by flow cytometry. Results The inhibition rate of tumor in PTX, PTL and PLTL groups was 48.87%, 57.22%and 78.87%, respectively. The apoptotic rate of tumor cell in PTX, PTL and PLTL groups was (42.7 ± 3 .8)%, (54.6 ± 2.9)%and (69.7 ± 5.0)%, respectively. Conclusions PLTL, as compared with PTX and PTL, has an evident thermo sensitive feature and increases the anticancer effect of paclitaxel remarkably in combination with local hyperthermia.

Objective To observe the effect of paclitaxel long-circulating thermo-sensitive liposome ( PLTL) on inhibiting the growth of transplanting Lewis lung cancer cells in C57BL/ 6 mice. Methods The model of mice carried Lewis lung cancer was established. 40 tumor-bearing mice were divided into five groups randomly: blank control group, model control group, PTX group, PTL group and PLTL group, 8 mice for each group. The blank control group and the model control group were injected with 0. 9% sodium chloride solution. In PTX group, PTL group and PLTL group, the dose of per injection was calculated with the reference of PTX for 20 mg·kg-1 , diluted with 0. 9% sodium chloride solution, and the mice were injected via the tail vein with a volume of 0. 2 mL each time. Except for the blank control group, 5 minutes after administration, the tumors of the other groups were subjected to local hyperthermia at (42±0. 5) ℃ for 30 min. During the treatment period, transplantation tumor growth was observed; pathological morphology changes of tumor tissues and cells were detected by HE stain; apoptosis rate of tumor cells was determined by flow cytometry to investigate the inhibition effect of PLTL combined with local thermal therapy on the tumor. Results The inhibition rate of tumor in model control group, PTX group, PTL group and PLTL group was 21. 81% , 48. 87% , 57. 22% and 78. 87% , respectively. The apoptosis rate of tumor cells was (20. 4 ± 4. 2)% , (42. 7 ± 3. 8)% , (54. 6±2. 9)% and (69. 7±5. 0)% , respectively. Observed by pathology, apoptosis rate and necrosis number of tumor cells in PLTL group were significantly increased. Conclusion Compared with PTX and PTL, PLTL has an evident thermo-sensitive feature and can increase the anticancer effect of paclitaxel remarkably when combined with local hyperthermia.

Objective To investigate the effect of combination therapy by observing the salivary glands function and related organ pathology after given methotrexate (MTX) and cyclophosphamide (CTX) intermittently in induced mice model of Sjogren's syndrome (SS). To further explore the synergistic effect of combination therapy by detecting the immunological regulatory factor B cell activation factor belonging to the TNF family (BAFF) expression. Methods The ingredients of Lewis rat's exocrine glands homogenate were injected into female C57BL/6 mice to set up the mice model of SS. After established the SS mice model successfully, they were randomly divided into six SS model group, including low-dose MTX treatment group (0.02 mg/w), high-dose MTX treatment group (0.06 mg/w), CTX pause treatment group (1.2 mg/3 w), CTX alternate day treatment group (0.6 mg/2 d), MTX+CTX combination treatment group (MTX 0.02 mg/3 w+ CTX 1.2 mg/3 w). Treatment effects were assessed both clinically and histologically. Results Eighteen weeks after the first treatment, the improvement of the salivary secretion of the CTX alternate day treatment group and MTX+CTX combination treatment group were higher than other groups, which showed statistically significant difference (P<0.01). Compared with the SS model control group, HE staining showed that the lymphocytic infiltration of exoerine glands was decreased in the treatment group. In the CTX alternate day treatment group and MTX+CTX combination treatment group, few amount of inflammatory cell infiltration were found, and the expression intensity of BAFF mRNA and protein were decreased markedly in salivary gland than others by RT-PCR and immunohistochemistry assay (P<0.01). Conclusion MTX and CTX can inhibit lymphocytic infiltration of the salivary glands, inhibit BAFF transcriptional level and production of BAFF protein, leading to an increase of fluid production. It suggests that modulation of signaling via BAFF pathways may be a mechanism of action. MTX and CTX combination therapy is more effective than single-agent therapy. The inhibitory effects of MTX and CTX on BAFF-mediated inflammatory pathways are primarily synergistic.

Objective To investigate the lipid profiles of patients with primary Sjogren's syndrome (pSS) and to analyze the correlation between abnormal serum lipids and the inflammationsof SS. Methods One hundred and fourteen pSS patients and 20 gendermatehed healthy controls were studied. Serum lipids were measured in both groups. Results There was statistically significant difference between SS and healthy controls, and the serum HDL-c and apoA<,1 concentrations were significantly lower in patients (P<0.05). The incidence of abnormal serum lipids was 39.5% in these patients. Patients with abnormal lipids had longer course of disease, higher ESR level, lower salivary flow rate and more frequent parotid gland enlargement than those without abnormal lipids(P<0.05). Logistic regression analysis revealed statistically significant association between serum lipids levels and occurrence of parotid gland enlargement. Conclusion Findings from this study suggest that patients with SS have altered lipid profiles and the decrease of apoA, and HDL-c levels may be the correlated factors of SS. The inflammation of SS may cause changes in lipids metabolisms.

Objective: To observe the effects of Na⁺ /H⁺ exchanger 1 (NHE1) inhibitor on intestinal injury of rats with burn sepsis, and to explore the possible mechanism preliminarily. Methods: Ninety SD rats were divided into control group, pure sepsis group, and NHE1 inhibitor group according to the random number table, with 30 rats in each group. Full-thickness scald (hereinafter referred to as burn) model with 20% total body surface area were reproduced on the back of rats in pure sepsis and NHE1 inhibitor groups, and then 50 μL liquid of Pseudomonas aeruginosa ATCC 27853 (2×10⁵ colony forming unit/mL) were injected into the center of wounds on the back. Rats in NHE1 inhibitor group were intraperitoneally injected with 0.1 mmol/L NHE1 inhibitor cariporide (0.4 mg/kg) rapidly after the successful establishment of burn sepsis model, while rats in pure sepsis group were injected with the same volume of normal saline. Except for not being made burn wounds nor receiving bacterination, rats in control group were treated the same as those in pure sepsis group. Rats with burn sepsis in each group were laparotomized and injected with 200 mL fluorescein isothiocyanate (FITC)-dextran in the concentration of 0.1 mol/L in terminal ileum at 12 hours post injury, and their left ventricular blood and terminal ileum were collected 30 minutes later. The serum content of FITC-dextran was detected with fluorescence spectrophotometer (n=10); the morphology of intestinal tissue was observed with HE staining (n=10); the content of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in serum and intestinal tissue was determined with enzyme-linked immunosorbent assay (n=20); the activity of myeloperoxidase (MPO) in serum and intestinal tissue was detected with colorimetric method (n=20); the protein expression of nuclear factor-kappa B-p65 (NF-κB-p65) and phosphorylation levels of mitogen-activated protein kinase (MAPK) signal pathway related proteins p38MAPK, extracellular signal-regulated kinase 1/2 (ERK1/2), and c-Jun N-terminal kinase 1/2 (JNK1/2) were determined by Western blotting (n=4). The same samples of rats in control group were collected for related detection at the same time point as above. Data were processed with one-way analysis of variance and SNK test. Results: (1) The serum content of FITC-dextran of rats in pure sepsis group was significantly higher than that in control group (P<0.01), while the serum content of FITC-dextran of rats in NHE1 inhibitor group was significantly lower than that in pure sepsis group (P<0.01). Compared with that in control group, infiltration of a large number of inflammatory cells, ulcer and necrosis of intestinal mucosa of rats in pure sepsis group were observed. The injury condition of intestine of rats in NHE1 inhibitor group was better than that in pure sepsis group. (2) The serum content of IL-6, TNF-α, and MPO of rats in pure sepsis group was (387±42) and (164.7±10.1) ng/mL, and (7.5±1.5) U/mL, respectively, significantly higher than that in control group [(75±17) and (13.1±6.5) ng/mL, and (2.3±0.7) U/mL, respectively, with P values below 0.01]. The serum content of IL-6, TNF-α, and MPO of rats in NHE1 inhibitor group was (176±37) and (64.9±9.3) ng/mL, and (5.9±0.8) U/mL, respectively, which was significantly lower than that in pure sepsis group (with P values below 0.01). (3) The content of IL-6, TNF-α, and MPO in intestinal tissue of rats in pure sepsis group was (190±13) and (172.8±29.7) ng/mL, and (8.7±1.5) U/mL, respectively, significantly higher than that in control group [respectively (20±3) and (11.9±2.3) ng/mL, and (2.9±0.3) U/mL, with P values below 0.01]. The content of IL-6, TNF-α, and MPO of intestinal tissue of rats in NHE1 inhibitor group was (35±6) and (45.2±6.1) ng/mL, and (5.3±0.6) U/mL, respectively, significantly lower than that in pure sepsis group (with P values below 0.01). (4) The protein expression of NF-κB-p65 and phosphorylation levels of p38MAPK and ERK1/2 in intestinal tissue of rats in pure sepsis group were significantly higher than those in control group (with P values below 0.01); the protein expression of NF-κB-p65 and the phosphorylation level of p38MAPK in intestinal tissue of rats in NHE1 inhibitor group were significantly lower than those in pure sepsis group (with P values below 0.01); phosphorylation levels of JNK1/2 in intestinal tissue of rats in the three groups were similar (with P values above 0.05). Conclusions The inhibition of NHE1 can significantly alleviate the intestinal injury, and the mechanisms may be attributed to the regulation of NF-κB and p38MAPK signal pathway, resulting in inhibition of the inflammatory response of intestinal tract.

Objective To evaluate the value of IgA and IgG antibodies against α-fodrin in both serum antibodies in SS is also assessed.Methods Samples from 39 patients with SS(25 primary and 14 secondary),8 patients with systemic lupus erythematosus(SLE),and 15 patients with rheumatoid arthritis (RA)as well as 10 healthy blood donors were collected.Anti-α-fodrin antibodies were measured using ELISA.Results The titer of serum anti-α-fodrin was higher in SS than in other connective tissue diseases group and healthy group(P<0.01).IgA type anti-α-fodrin antibodies was detected in 60%.44% of serum and saliva in patients with pSS respectively.IgG antibodies were detected in 43% of sera,and 29% of saliva of patients with pSS.The sensitivity and specificity of serum anti-α-fodrin IgA in SS was 54%and 85%.The level of anti-α-fodrin was positively associated with xerostomia and parotid swelling (P<0.05),and was negatively associated with xeroma,renal tubule acidosis,lung interstitial disease and hepatic damages(P>0.05).Conclusion Saliva and serLlm anti-α-fodrin level may be diagnostic for SS.It may be a useful screening marker.

MicroRNA (miRNA) is an endogenous, non-coding single-stranded RNA that regulates a variety of signal pathways or cytokines.Recent studies have confirmed that miRNA can affect alkaline phosphatase activity and matrix mineralization in bone formation, and plays an important role in osteogenic differentiation and cartilage differentiation.Abnormalities in the osteogenesis process can lead to osteogenesis imperfecta, Feingold syndrome, and femoral head necrosis.This review summarizes the specific mechanism of osteogenic differentiation and cartilage differentiation regulated by miRNA, suggesting the new clue for the future research about the underlying mechanism of bone development and clinical treatment of bone dysplasia from epigenetics.

Objective:To investigate the incidence, drug sensitivity and drug resistance characteristies, and theraputic effect of staphylococcal peritoneal dialysis-associated peritonitis (PDAP), aim to provide clinical evidences for standardizing treatment therapy of staphylococcal PDAP. Methods:Clinical data of PDAP patients admitted to the Second Hospital of Jilin University, the First Hospital of Jilin University-the Eastern Division, Jilin Central Hospital and Jilin First Automobile Work General Hospital during January 1, 2013 and December 31, 2019 were retrospectively collected. The results of etiology, drug sensitivity and drug resistance of staphylococcal PDAP patients were collected. According to the pathogenic bacteria, patients were divided into staphylococcus aureus group ( n=48) and coagulase-negative staphylococcus group ( n=232). According to the results of methicillin resistance, patients were divided into drug-resistant group ( n=71) and drug-sensitive group ( n=30). The prognosis of antibiotic therapy in each group were compared. Poisson regression was used to test the changing trend of the incidence of staphylococcal PDAP. The changes of drug sensitivity and drug resistance of staphylococcus were compared between 2013 and 2019 by linear trend χ2 test. Results:A total of 1 085 cases of PDAP occurred in 625 patients were screened, and 280 cases of staphylococcal PDAP were finally included. The incidences of staphylococcal PDAP, staphylococcus aureus PDAP and coagulase-negative staphylococcal PDAP were 0.063 times per patient year, 0.010 times per patient year and 0.053 times per patient year respectively. In addition, the incidence of PDAP caused by staphylococcus, staphylococcus aureus and coagulase-negative staphylococcus decreased year by year (all P<0.05). With the change of years, the sensitivity rate of staphylococcus to rifampicin increased, while the sensitivity rate of staphylococcus to moxifloxacin decreased (both P<0.05). The drug resistance rate of staphylococcus to levofloxacin increased ( P<0.05). The staphylococcus aureus group was more prone to refractory PDAP and catheter removal than that in coagulase-negative staphylococcus group, and the recurrence rate was higher than that in coagulase-negative staphylococcus group (all P<0.05). The proportion of vancomycin used during the whole course of antibiotic therapy in drug-resistant group was higher than that in drug-sensitive group ( P<0.05). Conclusions:The incidence of staphylococcal PDAP decreases year by year, and the drug sensitivity characteristics of staphylococcus also change. The therapeutic outcomes of staphylococcus aureus PDAP are worse than that of coagulase-negative staphylococcus.