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.

Histiocytosis, Langerhans-Cell ; Humans

Objectives To investigate the significance of electronic bronchoscopy and bronchoalveolar lavage in diagno-sis and treatment in children with pulmonary mass lesion. Methods A total of 74 hospitalized children from January 2011 to June 2012 whose imaging examinations showed massive patchy shadow were examined and treated by electronic bronchoscopy and bronchoalveolar lavage. Their clinical data were retrospectively analyzed. Results The major cause for the massive shadow was infection according to electronic bronchoscopy examination (68/74, 91.89%), 65 cases of them were lobar pneumonia, 3 cas-es were pulmonary tuberculosis followed by 5 cases of foreign body (6.76%) and one case of pneumorrhagia (1.35%).The lower left lung was the most frequently seen site of infection, followed by lower right lung. The agreement between infection sites and imaging examination was 97.30%. Bronchoalveolar lavage fluid showed that the primary pathogen of lobar pneumonia infection is Mycoplasma pneumoniae (MP) (42/65, 64.62%). The highest detection rate of MP was found in preschool group and the detec-tion rate between different age groups indicated statistically significant difference (P<0.01). The imaging examination showed pulmonary lesions in 61.54%children with lobar pneumonia improved significantly in one week. The improvement rate of pul-monary lesions was higher in infected children with short duration (1-2 weeks, 90.91%) between disease onset and electronic bronchoscopy inspection than those with longer duration (2-3 weeks, 51.72% and >3 weeks, 35.71%) (P<0.05). Conclusions Electronic bronchoscopy and bronchoalveolar lavage play dual roles in etiological diagnosis and therapy in children with pulmo-nary mass lesion.

Objective To investigate whether the release of fibroblast growth factor-1 ( FGF-1 ) was changed after inhibition of S100A13 gene (small hairpin RNA, shRNA)and serum-deprivation in human thyroid cancer cells (TT cells ). Methods The S100A13-shRNA pENTRTM/U6 entry vector was transfected into TT cells. The expression of S100A13 mRNA and protein was detected by immunoflurescence, real-time RT-PCR, and Western blot. Then TT cells were treated with S100A13 gene inhibition and serum-deprivation. The changes in release of FGF-1 were detected by indirect immunoflurescence, RT-PCR, and ELISA. Results S100A13 shRNA transfected TT cells (S100A13 RNAi cells)had a reduction of S100A13 gene and protein expression by 80%.Indirect immunofluorescence indicated FGF-1 was mostly localized in the cytoplasm and nucleus of TT cells in primary culture. When serum-deprivation stress was given to TT cells, FGF-1 in cytoplasm almost disappeared in the cells at 6 h. RT-PCR indicated that when serum-deprivation stress was given to TT cells the mRNA of FGF-1 was reduced. ELISA showed that with inhibition of S100A13, the release of FGF-1 was reduced (P＜0.05).Conclusion S100A13-shRNA pENTRTM/U6 entry vector transfected TT cells may inhibit the expression of S100A13 and reduce the release of FGF-1.

There are some problems with this course of medical laboratory special English, such as reference materials, teaching contents, teaching ways, the evaluation forms, etc. The department of medical laboratory of North Sichuan Medical College did exploratory reforms including 1+n teaching mode (one major teacher and several co-operational teachers in discussion section), Flipped classroom and interactive teaching, new formative assessment forms (usual performance combing final shows grade), etc. They used innovative teaching way with the purpose of establishing new teaching way , cultivating abilities of au-tonomous learning and comprehensive application.

Objective Irregular antibody screening and identification are very important to reduce hemolytic reaction.The dis-tribution of irregular antibodies of blood group was analyzed in patients, to provide evidence for using Micro column gel test to detect and identify irregular antibodies transfusion safety. Methods Micro column gel anti-globulin technique was used to screen the irregular an-tibodies and cross-matching in 29770 patients.If the serum had irregular antibodies, the spectrum cells were used to identify the specific-ity of irregular antibodies. Results Among 29 770 patients, 120 cases had irregular antibodies of blood group.The positive rate was 0.4%.The irregular antibodies include Rh system(n=67),MNS system (n=7), Lewis system(n=5),cases of Kidd system(n=3) and undefined specified antibodies(n=38).Among the 120 antibody-positive patients, 70 were female patients and 50 were male patients. Further study showed that the pregnancy history was to the main factor causing irregular antibodies in women (46 positive cases among 70 female patients) . Conclusion The study showed that some patients had irregular antibodies of blood group, especially female patients with pregnancy history.These results suggested that irregular antibody screening and cross-matching by micro column gel method in pa-tients before transfusion had great significance for ensuring the blood transfusion safety.

OBJECTIVE The aim of the present study was to evaluate the protective effects of momordica charantia polysaccharides(MCP)on depressive animal model induced by chronic social defeat stress(CSDS)and explore the underlying mechanisms.METHODS We established CSDS depressant mouse model and treated CSDS mice with MCP.Sucrose preference,forced swim test(FST)and social interaction test(SIT)were used to measure behaviors changes.We used ELISA,Q-PCR and western blot to test the levels of cytokines in the hippocampus. RESULTS The results showed that chronic administration of MCP(100,200 and 400 mg·kg-1)significantly prevented depressive-like behaviors in mice as assessed by social interaction (SIT), tail suspension (TST) and sucrose preference tests (SPT).It was showed that the elevation of proinflammatory cytokines(TNF-α,IL-6,and IL-β)concentra-tions,up-regulation of JNK3,c-Jun,and P-110β protein expressions in the hippocampus of CSDS model. Moreover,reduction activity of PI3K and phosphorylation level of protein kinase B(AKT)was also observed in the hippocampus of CSDS model.All above phenomenon were reversed after MCP intervened.Further-more,the protective effects of MCP on the CSDS mice were partly inhibited by the specific phosphati-dylinositol 3-kinase (PI3K)inhibitor, LY294002. CONCLUSION The protective effects of MCP against depressive-like effects in CSDS mice might reduce neuroinflammatory and involve in attenuation of JNK3/PI3K/AKT pathway in the hippocampus.

Objective:To observe the clinical effect of mild moxibustion for knee osteoarthritis (KOA) and to explore the role of mild moxibustion in relieving pain,reducing stiffness and improving joint dysfunction in patients with KOA.Methods:Eighty patients with KOA were randomly allocated into either a moxibustion group or a medication group by the random number table,with 40 cases in each group.The moxibustion group used mild moxibustion at Neixiyan (EX-LE 5),Dubi (ST 35),Xuehai (SP 10) and Liangqiu (ST 34),30 min each time,3 times a week;the medication group was given celecoxib capsule (celebrex),0.2 g each time,once a day.Both groups were treated for 4 weeks.The visual analog scale (VAS) and Western Ontario and McMaster Universities osteoarthritis index (WOMAC)scores were evaluated before and after treatment.The efficacy of the two groups was compared after treatment.Results:After treatment,the overall efficacy of the moxibustion group was significantly different from that of the medication group (P＜0.05).The VAS and WOMAC scores of the two groups were lower than those before treatment (both P＜0.01).The changes in the VAS and WOMAC scores after treatment in the moxibustion group were significantly different from those in the medication group (both P＜0.05).After treatment,in single item of WOMAC,the changes in pain and joint dysfunction in the moxibustion group were more statistically significant than those in the medication group (both P＜0.05).Conclusion:Mild moxibustion and oral celebrex can reduce the VAS and WOMAC scores of patients with KOA.Mild moxibustion is superior to oral celebrex in relieving pain and improving joint function.

OBJECTIVETo investigate the changes of vasoconstriction and vasodilatation under different temperature conditions and the protective effects of Vitamin E (Vit E) against endothelial injury induced by hypothermia.

METHODSThe tail arterial rings were prepared for isometric tension recording using multi wire myograph system. The effect of temperature on relaxation and construction was evaluated. Incubate the arterial rings with different concentration of Vit E when they were exposed to hypothermia, then acetylcholine (ACh)-induced endothelium-dependent relaxation was investigated to evaluate the activity of endothelial.

RESULTS(1) The hypothermia could enhanced the dose-dependent construction induced by PE in mice tail artery. (2) Exposure to hypothermia also resulted in increase of sodium nitroprusside (SNP)-induced re-After incubation with Vit E, the vascular relaxation responses to ACh increased in an endothelium-dependent manner, when compared with the hypothermia-treated group.

CONCLUSIONThe vascular function of constriction was attenuated by hypothermia, while the relaxation was increased. Vit E could prevent the hypothermia-induced decrease in vascular endothelial cells.

Animals ; Arteries ; drug effects ; physiology ; Cold Temperature ; Hypothermia ; In Vitro Techniques ; Male ; Mice ; Prazosin ; pharmacology ; Solanaceous Alkaloids ; pharmacology ; Vasoconstriction ; drug effects ; Vasodilation ; drug effects ; Vasodilator Agents ; pharmacology ; Vitamin E ; pharmacology