MiRNA is a small non-coding single stranded RNA(ssRNA)which is made up by 21~23 nucleotide.This ssRNA has a double effect as tumor suppressor genes and oncogenes , and plays an important role in cell proliferation , differentiation and apop-tosis.Pancreatic carcinoma is a highly malignant tumor with poor prognosis and high mortality rate , and the early diagnosis is difficult . In the last decades , growing studies highlighted the interactions between miRNAs and pancreatic carcinoma .This review provides a re-search overview between miRNAs and pancreatic carcinoma .Meanwhile , the significance of Expression Profile in pancreatic carcinoma′s diagnosis ,treatment and prognosis is also discussed .

Objective To study the complexity of S region qusispecies in various disease stages of chronic hepatitis B virus(HBV) infection and its relation to disease activity. Methods Serum samples were obtained from 112 patients with chronic hepatitis B virus infection;22 with chronic carries(ASC),30 with chronic mild or moderate hepatitis(CH),60 with fulminant hepatitis failure(FHF). HBV qusispecies populations were separated by the single strand conformation polymorphism (SSCP) method targeted the S region and DNA sequencing analysis. Results The number of SSCP bands detected in the patients with ASC、CH and FHF was 1.45?0.13,3.70?0.22 and 5.93?0.24, respectively. There was a statistically significant difference in the number of quasispecies among various disease stages ( P

Objective To investigate difference of NKG2D receptor expression level on the surface of natural killer (NK) cells in the patients with hepatitis B and its clinical significance.Methods This was a four-arm study with different types of subjects,including patients with chronic hepatitis B (CHB,n =22),HBV carriers (HBVC,n=10),patients with acute hepatitis B (AHB,n=18) and healthy donors (HD,n=18).NKG2D protein and mRNA levels on the surface NK cells in the peripheral blood were examined by reverse transcription-polymerase chain reaction assay.The relationship between NKG 2D expression and serum hepatitis B virus (HBV) DNA level was analyzed.The data were compared by analysis of variance and linear regression.Results NKG2D mRNA expression levels in groups of HBVC, HD, AHB and CHB were 0.96±0.17, 1.03±0.12,1.53±0.30 and 1.51 ± 0.35,respectively; the differences among groups were statistically significant (q=7.586,7.485,7.920 and 7.880,respectively; all P＜0.01).NKG2D protein expression levels in groups of AHB,HD,CHB and HBVC were 0.87±0.14,0.89±0.17,0.67±0.09 and 0.59±0.13,respectively; the differences among groups were statistically significant (q=6.92,7.67,7.53and 8.16,respectively; all P＜0.01).The NKG2D mRNA expression levels on NK cells were negatively correlated with serum HBV DNA viral loads in patients with CHB,AHB or HBVC (r=-0.75,-0.66 and-0.69,respectively; all P＜0.01).The NKG2D protein levels on NK cells from patients with AHB and CHB were negatively correlated with serum HBV DNA levels (r=-0.47 and -0.45,respectively; both P＜0.05).Conclusion NKG2D mediated NK cytotoxicity may play a role in viral clearance in hepatitis B.

AIM:To investigate the target killing effect of T lymphocytes with chimeric CD20scFv gene on Daudi cells and the activation of T lymphocytes.METHODS:Two kinds of plasmids were transfected into retrovirus-packed PA317 cell lines.The supernatant was collected from successfully transfected PA317 culture and was used to infect peripheral blood T lymphocytes.After one-week screening with G418,the cells were used to kill Daudi and K562 cells.The positive rates of AnnexinⅤ in Daudi cells were measured at different times points respectively by flow cytometry.Meanwhile,the level of IL-2 and IFN-? were determined by ELISA.RESULTS:The Annexin V positive rate was significant higher in Daudi cells compared to control K562 cell lines at 24 h.No difference of AnnexinV in Daudi cells was observed in CD20 modification T lymphocyte groups.The secretions of IL-2 and IFN-? in CD20scFv-CD80-IgGFc-CD28-? gene modified T cells co-cultured with Daudi cells were dramatically higher than that in CD20scFv-IgGFc group at 72 h.CONCLUSION:① The two kinds of genetic modified specific T cells have no significant difference in inducing early apoptosis of Daudi cells.CD28-? can't affect Daudi cell early apoptosis at the CD20scFv target killing.② The increase in the secretions of IL-2 and IFN-? is more obvious in CD20scFv-IgGFc-CD28-? group,indicating that the self-activation takes place in CD3? and CD28 modified T cells without MHC restriction and then increases the activation and killing function of T cells.

Objective To investigate the effect of cannabinoid receptor 1 ( CBR1 ) on spatial learning and memory function of neuropathic pain ( NP ) model rats and the expression of N-methyl-D-aspartic acid receptor 1(NR1) subunit in medial prefrontal cortex (mPFC).Methods Thirty-six healthy male Wistar rats were randomly divided into 4 groups, with 9 rats in each group: the sham operated group (SO group), the neuropathic pain model group (NP group), the NP model group with an mPFC injection of saline ( NS group ) , and the NP model group with an mPFC injection of the CBR 1 antagonist AM251 ( AM251 group).The NP model was prepared using the operation of chronic constriction injury ( CCI) of the right sciatic nerve.The mechanical withdrawal threshold ( MWT ) and the thermal withdrawal latency (TWL) of the rats in each group were detected at 3, 7, 14, 21 and 28 days after the operation.At 29 days after the operation , 18 rats of NP model were randomly selected and given an mPFC injection of saline or AM251 using a three-dimensional brain puncture.At days 30-37 after operation , the eight-arm maze test was performed to detect the spatial learning and memory function of the rats , and the rats were sacrificed immediately after this test.The expression levels of CBR1, NR1 and phosphorylated-N-methyl-D-aspartic acid receptor 1 ( p-NR1 ) ( Ser896 ) in the mPFC brain region were detected by Western blotting , RT-PCR and immunofluorescence.Results Compared with the SO group , the pain thresholds and the spatial learning and memory function of the rats in the NP group were significantly lower ( both P <0.05 ).Compared with the NS group , the rats in the AM251 group showed improvement about spatial learning and memory function ( P<0.05).Compared with the SO group ( the mRNA and protein level of CBR 1:0.23 ± 0.06,0.42 ±0.03), the mRNA(0.43 ±0.12) and protein (0.53 ±0.05) level of CBR1 in NP group increased (both P<0.05).Compared with the NS group (the mRNA and protein level of CBR1:0.42 ± 0.11,0.52 ±0.10), the mRNA (0.53 ±0.05) and protein (0.98 ±0.17) level of CBR1 in AM251 group increased (both P<0.05).Compared with the SO group (the mRNA and protein level of NR1 and the protein level of p-NR1:1.50 ±0.15,0.65 ±0.05,0.79 ±0.15), the mRNA (0.94 ±0.07) and protein (0.24 ±0.05) level of NR1 in NP group decreased (both P<0.05), the protein level of p-NR1 (0.33 ± 0.04) decreased (P<0.05).Compared with the NS group (the mRNA and protein level of NR1 and the protein level of p-NR1:1.09 ±0.14,0.26 ±0.06,0.31 ±0.08), the mRNA(1.58 ±0.10) and protein (1.42 ±0.10) level of NR1 in AM251 group increased (both P<0.05), the protein (0.95 ±0.15) level of p-NR1 increased ( P<0.05).Conclusion CBR1 can decrease the expression level of NR 1 and p-NR1 in the mPFC brain region of NP model rats and induce the spatial learning and memory impairment.

Objective To investigate the protective effect of fibrauretin injection against acute lung injury induced by lipopolysaccharide (LPS) in mice.Methods Seventy-two healthy male adult Kunming mice were randomly divided into 6 goups:normal control group,LPS group (intratracheal instilation of 5 mg/kg of LPS),hydrocortisone group (intraperitoneal injection of 3.3 mg/kg of hydrocortisone,once daily for 3 days,and intratracheal instilation of 5 mg/kg of LPS on the 4th day) and low dose,medium dose and high dose of fibrauretin groups (intraperitoneal injection of 2,10 and 50 mg/kg of fibrauretin,respectively,once daily for 3 days,and intratracheal instilation of 5 mg/kg of LPS on the 4th day).The mice in each group were sacrificed by dislocation 24 h after intratracheal instilation of LPS.The lung tissues of partial mice in each group were extracted and weighed to calculate the lung coefficients,and bronchoalveolar lavage (BAL) was performed in partial mice in each group to collect the BALF for counting the inflammatory cells.Results The lung cofficients of mice in LPS group were significantly higher than those in the normal control group (P＜0.05).The lung cofficients of mice in hydrocortisone group,low dose,medium dose and high dose of fibrauretin groups were significantly lower than those in LPS group (P ＜ 0.05).For the percentage of inflammatory cells in BALF and the percentage of neutrophils in inflammatory cells,LPS group was significantly higher than the normal control group (P＜0.01),and compared with LPS group,hydrocortisone group,low dose,medium dose and high dose of fibrauretin groups were significantly decreased (P＜0.01).Conclusion Fibrauretin injection can significantly ameliorate the inflammatory reaction degree in mice with acute lung injury induced by LPS.

AIM:To construct a recombinant eukaryotic expression vector pLNCX/anti-CD20scFv/IgGFc/CD80/CD28/? and detect its expression in NIH 3T3 cells.METHODS:CD28-? cDNA was amplified from the plasmids pBULLET and inserted into pLNCX vector that contained anti-CD20 scFv/IgGFc/CD80 gene.The recombinant plasmids were transfected into NIH 3T3 cells,and resistant clones were obtained by G418 selection.The gene expression of the fusion protein was determined by RT-PCR and FACS.RESULTS:The recombinant eukaryotic vector was constructed successfully,determined by PCR and enzyme digestion analysis.The target gene was amplified from NIH 3T3 cells transfected with the vectors by RT-PCR.The FACS showed that recombinant protein was expressed in NIH 3T3 cells.CONCLUSION:Construction of pLNCX/anti-CD20scFv/IgGFc/CD80/CD28/? expression vector and its expression in NIH 3T3 cells lay the foundation for further research of generation of modified T lymphocytes to CD20 positive lymphoma.

Objective: To evaluate the clinical value of the plasma microRNA-155 (miR-155), miR-196a, miR-21 and miR-210 in early diagnosis of patients with pancreatic cancer. Methods: The real-time fluorescent quantitative-PCR was performed to detect the levels of miR-155, miR-196a, miR-21 and miR-210 in plasma samples from sixty patients with pancreatic cancer, twenty patients with chronic pancreatitis, and ten healthy volunteers, as well as the levels of four miRNAs in cancer tissue specimens from ten patients with pancreatic cancer. The serum tumor markers carbohydrate antigen 199 (CA199), CA242 and carcino-embryonic antigen (CEA) were simultaneously detected. The relative expression levels of four miRNAs in plasma among these three groups were compared, and the relationship between miRNA levels in plasma and their levels in pancreatic cancer tissues were statistically analyzed. Finally, the diagnostic efficiency of plasma miR-155, miR-196a, miR-21 and miR-210 for pancreatic cancer was evaluated. Results: The relative expression levels of plasma miR-155, miR-196a, miR-21 and miR-210 in pancreatic cancer group were significantly higher than those in the chronic pancreatitis group and the healthy control group (all P < 0.01). The relative levels of miR-155, miR-196a, miR-21 and miR-210 in pancreatic cancer tissues were higher than those in plasma from the same patients with pancreatic cancer, but there were no statistically significant differences (all P > 0.05). The area under receiver operating characteristic (AUC-ROC) curve of plasma miR-155, miR-196a, miR-21 and miR-210 in diagnosis of pancreatic cancer indicated that these four miRNAs had diagnostic value independently (all P < 0.01), in which miR-155 had the highest diagnostic efficiency. The binary Logistic regression model showed that combined detection of plasma miR-196a and miR-210 was more effective in diagnosis of stageI pancreatic cancer as compared with CA199. Conclusion: The plasma miR-155, miR-196a, miR-21 and miR-210 levels are effective to distinguish pancreatic cancer from non-pancreatic cancer. The combined detection of miR-196a and miR-210 may become a promising method for early diagnosis of pancreatic cancer.

Most early-stage cervical cancer patients achieve good recovery through surgical treatment and concurrent chemoradiotherapy. However, for patients with recurrent, metastatic cervical cancer, the available effective treatment is rare and the prognosis is poor. In recent years, with the development of immunotherapy, especially immune checkpoint inhibitors targeting programmed death-1 (PD-1) and its ligand (PD-L1) and cytotoxic T-lymphocyte associated protein-4 (CTLA-4) , such as pembrolizumab, nivolumab, ipilimumab, has made breakthrough progress in the treatment of recurrent or metastatic cervical cancer.

Objective To analyze the composition and influencing factors of the hospitalization expenses of patients with cardiovascular and cerebrovascular diseases who has participated in basic medical insurance, and to provide evidence for controlling excessive increase in the hospitalization expenses and reducing the financial burden of patients. Methods The hospitalization information of 14,271 insured patients with cardiovascular and cerebrovascular diseases from January 1, 2019 to December 31, 2019 in Xianning City, Hubei Province was retrospectively collected. The basic information of the patients and the composition of their hospitalization expenses were descriptively analyzed, and the influencing factors of hospitalization expenses of the patients were analyzed by univariate analysis and logistic regression analysis. Results Among the patients included in the study, coronary heart disease, cerebral infarction, cerebral hemorrhage and essential hypertension were the four main types of cardiovascular and cerebrovascular diseases with the largest proportion of hospitalization expenses, accounting for 26.18%, 20.29%, 11.82% and 9.94%, respectively. The largest proportion of hospitalization expenses was treatment expenses and drug expenses, accounting for 44.09% and 32.52%, respectively. Logistic regression analysis showed that age, length of stay, type of insurance, type of cardiovascular and cerebrovascular diseases, whether there were other comorbidities or complications, and whether they visited tertiary medical institutions were the influencing factors of hospitalization expenses for patients with cardiovascular and cerebrovascular diseases. Conclusion It is necessary to strengthen the disease prevention and control for the elderly and patients with cardiovascular and cerebrovascular diseases such as coronary heart disease, cerebral infarction, cerebral hemorrhage and essential hypertension, accelerate the integration of the basic medical insurance system, scientifically and reasonably shorten the length of hospital stay, and strengthen the promotion of the hierarchical medical system.