Stomatology is an applied science,and experimental teaching plays an important role in training renovation talents.A complete construction of the laboratory system is a complex systematic project,which requires the right teaching conception,advanced experimental instruments,high-quality teachers of laboratory,the optimization of the teaching contents,as well as highly efficient operating mechanism and management approach.It is also a process of exploration and innovation.

Objective To select microRNAs (miRNA) associated with human cutaneous malignant melanoma (MM). Methods Total RNA was extracted from 6 tissue samples of MM and 9 human control samples of pigmented nevi, and small RNAs of less than 200 bp were enriched, miRNA microarray was used to select differentially expressed miRNAs between tissue samples of MM and pigmented nevi from 468 candi-dates. The expression of differentially expressed miRNAs was confirmed by fluorescence based real-time quan-titative PCR (qPCR) in all of these samples. Those miRNAs that were identified as differentially expressed with both miRNA microarray and qPCR were considered as significant miRNAs. Results Between the tissue samples of MM and pigmented nevi, 12.18% to 86.33% of miRNAs differentially expressed by more than 2 folds, 1.28% to 19.02% by more than 5 folds, and 0.43% to 5.34% by more than 10 folds. The expression of miRNA-21 was obviously up-regulated, while that of miRNA-320 and miRNA-494 was down-regulated in the MM samples. Conclusion There is an increase in the expression of homo sapiens miRNA-21 but a decrease in that of miRNA-320 and miRNA-494 in MM tissues.

Animals ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis ; metabolism ; virology ; Myocardium ; metabolism ; Osteopontin ; metabolism ; Virus Diseases ; metabolism

ABSTRACT:Objective To construct V-set and immunoglobulin domain containing 4 (Vsig4)nanobodies (Nbs) as specific macrophage probes so as to use them as molecular probes of macrophagocytes.Methods A nanobody phage library was generated by using peripheral blood lymphocytes isolated from an alpaca immunized with recombinant Vsig4 protein.After three rounds of selection against recombinant Vsig4.The Nbs were subjected to sequencing and genome alignment to obtain VHH sequence.Nbs were isolated and tested for Vsig4 specificity in an ELISA using recombinant Vsig4.The affinity capacity of Nbs was verified by the cell line stably expressing Vsig4. Results A nanobody phage library with an estimated 7.27 × 107 clones with 70% insertion was successfully constructed.Totally 1 3 6 Vsig4-positive clones were sequenced and aligned according to different CDR3 sequences. In summary,1 5 Vsig4 nanobodies were obtained and grouped into 3 different CDR3 epitopes.The affinity of representing nanobody and Vsig4 was analyzed via ELISA;Nb1 1 9 showed the highest affinity against both recombinant and native Vsig4.Conclusion We successfully constructed and screened Vsig4 specific nanobody number 1 1 9 with high affinity and specificity.It can help with macrophage detection and in vivo monitoring.

Immunologic classification of acute lymphocytic leukemia (ALL) was carried out by using 11 - 13 kinds of monoclonal antibodies against human leucocyte differentiation antigens in 19 cases. The results showed that of these 19 cases, there were T-ALL in 4, B-ALL in 4, C-ALL in 7, AUL in 2 and hybrid type of ALL (H-ALL) in 2. The 5 types mentioned above could also be classified, according to the immunologic criteria described by Foon in 1986, into 3 major groups: T-ALL, non-T-ALL and H-ALL. The Foon's classification method is useful for judgement of malignant cell source and differentiated stage, and for further understanding the nature of ALL cells.

Objective To explore the value of tubercular antibody (TBAb) in the diagnosis of tuberculous pericarditis (TBP).Methods The TBAb in 38 patients were determined by colloidal gold (CG) and compared with 64 healthy control. Results The positive ratio in TBP group was significantly higher as compared with control group. The hydropericardium was significantly decreased or disappeared after 6 months of antituberculotic treatment by color Doppler ultrasonography. It seemed that patients were rehabilitated and electrocardiogram (ECG) was normal. Conclusion TBAb is valuable in the diagnosis,differential diagnosis or treatment of TBP.

Objective To observe the clinical efficacy of Xuanfeitongfu method in treatment of severe pneumonia and to explore its clinical value in the management of severe pneumonia. Methods Totally,62 patients with severe pneumonia were randomly divided into the control group (n = 30) and the treatment group (n = 32). The control group was given cluster treatment,including oxygen cure,anti-infection and nutrition support and maintaining a stable internal environment and etc. The treatment group was treated with Tongfu decoction orally(one dose a day,a total of 5 days)on a basis of cluster treatment. The comparison was conducted in the 2 groups in the levels of C reaction protein,calcitonin,the change of blood gas analysis,the time of mechanical ventilation and the mortality of severe pneumonia in ICU at baseline and 3,5 days after treatment. Results The level of C-reactive protein,calcitonin,the change of blood gas analysis were statistically significant (P < 0.05). The time of mechanical ventilation and the mortality were better in the treatment group (P < 0.05). Conclusion Xuanfeitongfu method can effectively improve oxygenation ,assist the anti-infection effect ,reduce the time of mechanical ventilation and ICU retention time ,ultimately improve the outcome of severe pneumonia.

Objective To observe the therapeutic effects of nerve blockade combined with millimeter wave on patients with head acute herpes zoster (HZ) following serious pain.MethodsForty-two patients with head acute HZ following serious pain were randomly assigned to two groups: the combined group which treated with nerve blockade combined with millimeter wave, and the control group which received only nerve blockade. The pain degree (Numeric Rating Scales, NRS), blisters and scabbing were evaluated after treatment.ResultsNRS significantly decreased in both groups after treatment. The blister was earlier to disappear and scabbing was better in the combined group than in the control group (P<0.01).ConclusionNerve blockade combined with millimeter wave can effectively relieve the pain and shorten course of head acute HZ following serious pain.

Objective To investigate the expression of microRNA-21(miRNA-21)in two human malignant melanoma cell lines,A375 and M14,and its effect on the viability of these cells.Methods The expression of human miRNA-21 was assessed by quantitative fluorescent PCR in A375 and M14 cells.Then,three concentrations(90,180,270 nmol/L)of miRNA-21 inhibitor and were transfected both cells and a negative control were transfected a mixture of LipofectamineTM 2000 reagent,respectively.After another 3-day culture,the proliferation of cells was detected by cell counting kit-8,and R value was calculated to denote the relative activity of cells.Statistical analysis was carried out by SPSS13.0.Results The expression of miRNA-21 was higher on A375 cells than that on M14 cells with the average value of 2-deltaCT being 1.2928±0.1509 vs 0.1894±0.1803.With miRNA-21 inhibitor at the concentration of 90,180,270 nmol/L,the activity of A375 cells was significantly lowered in comparison with that in the control group,with the R value being 0.7362±0.1662.0.7248±0.3204 and 0.6767±0.2998 respectively(all P＜0.01).However,in the case of M14 cells,cell activity was only suppressed by miRNA-21 inhibitor at 90 nmol/L with the R value being 0.7295±0.1478.and no significant inhibition was observed with the inhibitor at 180 or 270 nmol/L (both P＞0.05).Conclusions miRNA-21 is expressed on human melanoma cell lines,A375 and M14,at different levels,with a promoting effect on the proliferation of both cells.Moreover,miRNA-21 may act as an oncogene-like gene via down-regulating the expression of some tumor-inhibiting factors.

Objective To evaluate the efficacy and safety of autologous hematopoietic stem cell transplantation (AHSCT) in patients with relapsed and refractory malignant lymphoma. Methods The clinical data of 10 patients (6 males and 4 females) with relapsed (4 cases) and refractory (6 cases) malignant lymphomas who received AHSCT in General Hospital of Jinan Military Command from August 2011 to June 2015 were analyzed retrospectively. The median age was 34 years (20 ˉ50 years); 5 cases of Hodgkin lymphoma, 5 cases of non-Hodgkin lymphoma. Before transplantation, all patients received several courses of radiotherapy or chemotherapy. High dose of cytoxan combined with G-CSF were used to mobilize peripheral hematopoietic stem cell, and preconditioning regimen included BEAM, CBV or TBI. Results The median mononuclear cell of 10 patients was 7.385 × 108/kg. Complete remission was achieved in 8 patients after transplantation, and 2 cases relapsed. Median follow-up time was 18 months (20ˉ50 months). The overall survival rate and disease-free survival rate both were 80%(8/10). All patients had nausea, vomiting, diarrhea, oral mucositis and other adverse reactions, which could be tolerated. Conclusion ASHCT is an effective and safe method for treatment of relapsed and refractory malignant lymphomas.