Objective To construct the expression plasmid of porcine leukemia inhibitory factor(LIF) in the prokaryotic system,and purify the expressed recombinant porcine LIF protein as antigen to immune rabbit for preparation of rabbit anti-pig LIF polyclonal antibody,and immunolocalize of LIF in pigs.Methods The cDNA fragment encoding mature porcine LIF protein was amplified by PCR,then cloned into the prokaryotic expression vector PET-31b at restriction sites NdeⅠand XhoⅠ.After transformed into E.coli BL21 for expression,the recombinant porcine LIF protein was purifed for preparation of anti-pig LIF polyclonal antibody as immunogen.Western blotting and ELISA were used to test the specificity and titer of the purifed IgG.Immunostaining was used to assay the expression of LIF in pigs.Results The constructed expression plasmid was identified by DNA sequencing and restriction enzyme digestion.The expression of the recombinant porcine LIF protein was performed exactly in E.coli BL21.By immuning the rabbit,the polyclonal antibody was successfully prepared.The results of ELISA and Western blotting showed that the antibody had high titer(1∶10000) and specificity.By immunostaining,LIF protein was faintly expressed in porcine myocardial cell,splenic cord and monolayer covering epithelium cell and pulmonary fibroblast.Conclusion The prepared polyclonal antibody has high titer and specificity;LIF has faint positive immunoreactivity in the normal porcine,myocardial cell,splenic cord and monolayer covering epithelium cell and pulmonary fibroblast.

AlM: To study the trabeculectomy clinical effect of use tunnel knife to make double - deck scleral flap and to cut off the layer scleral flap of glaucoma.METHODS: Using the random grouping method to divide 46 cases (60 eyes) of glaucoma into the treatment group of 24 cases (32 eyes) and control group of 22 cases (28 eyes). The treatment group, tunnel knife was used to make double- deck sclera flap and superficial scleral flap about the size of 5mm×5. 5mm, 1/3 scleral thickness, under the sclera flap made another one about the size of 3. 5mm× 4mm, 1/3 scleral thickness, resected the middle layer of the sclera flap, removed 2mm×2mm trabecular tissue, underwent routine peripheral iridectomy, could adjust suture the superficial scleral flap, sutured Ball fascia and bulbar conjunctiva. ln control group, routine glaucoma trabeculectomy was undergone.RESULTS:Patients were followed up for 1a, the vision in treatment group was obviously better than that in the control group, with a statistically significant difference (P< 0. 05). The postoperative intraocular pressure of the two groups of patients were significantly lower than that of the preoperative one. Postoperative 1 and 3mo, no statistical significant difference of intraocular pressure in two group(P>0. 05). But after 6 and 12mo, the intraocular pressure of the treatment group were significantly lower than that of the control group, with statistically significant difference (P<0. 05). Postopeartive 1a, the cumulative complete success rate and conditions for successful rate were 90. 63% and 96. 88% in the treatment group, and those were 75% and 89. 29% in control group. There was significant difference between two groups(P<0. 05).CONCLUSlON:The trabeculectomy have a good effect to lower the intraocular pressure by use tunnel knife to make double-deck scleral flap and to cut off the layer scleral flap. The scleral flap have uniform thickness, smooth surface, and the function of the filtering bleb maintained for a long time, less postoperative complications, suitable for various types of glaucoma, so it is worthy of clinical promotion.

Objective:To investigate the effects of dendritic cells(DCs)in pathogenesis of aplastic anemia,especially its roles in T cell abnormal activation.Methods:Direct immunofluorescence assays and flow cytometry(FCM)were used to determined proportion and kinds of DCs in bone marrow sample of patients with AA.The DCs were got from standard method by culturing mononuclear cells isolated from peripheral blood of normal person in medium with recombinant human granulocyte macrophage colony stimulating factor(rhGM-CSF)and interleukin 4(IL-4)in vitro.The phenotype of DCs were determined by FCM.After pulsed with purified cord blood CD34+ cells and stimulated with recombinant human CD40 ligand(rhCD40L),the ability of those cultured CDs to activate and promote auto T lymphocytes to proliferation was tested.The changes of TCR V? gene repertorie of those T cells were also investigated by real time quantitative PCR(RQ-PCR).Results:The CD1a+CD11c+ and CD11c+CD83+ double positive cells increased in bone marrow of patients of AA,especially the proportion of CD11c+CD83+ cells increased significantly in those patients.After primed with CD34+ cells,DCs induced from peripheral monocytes could activate T lymphocytes and promote T cells to proliferation.The restricted usage of TCR V? genes was confirmed by RQ-PCR in those activated T cells by DC as those in AA patients.Conclusion:DCs,especially with phenotype of CD11c+CD83+,may have great impact on oligoclonal expansion of lymphocytes in patients AA,but the antigen which recognized by auto T cells and stimulated it to proliferation are still needed to further investigation.

Adult ; Hand Injuries ; surgery ; Humans ; Male ; Occupational Diseases ; surgery ; Paint

TheStandardized Manipulations of Acupuncture and Moxibustion has been published and gradually standardized the clinical practice. It provided a solid foundation for the standardization of acupuncture and moxibustion therapeutic effects. It also provided powerful convictable effects on the accuracy and scientificity of acupuncture and moxibustion clinical practice, which gained well-acceptance from patients. In the treatment of certain diseases, acupuncture and moxibustion have become irreplaceable so that acupuncture and moxibustion can be strongly promoted both at home and abroad. There were some common problems occurred during the establishment of the national standardization onStandardized Manipulations of Oral and Lip Acupuncture. This article reviewed, analyzed and discussed questions occurred during the standardization establishment. The results showed that according to the principle of standardized production, a rigorous, scientific and reasonable text standard can be established. Advices or opinions gained during the process can be used as references for future standardization establishment. It was concluded that the standardization of acupuncture had great guidance in the acupuncture clinical application. It is an arduous and long-term mission to achieve and strengthen the position of international standardization of acupuncture and moxibustion. The government should increase the effort on investment and promotion of acupuncture and moxibustion standardization. Conversation and cooperation should also be strengthened with international communities to promote the standardization implementation.

Objective To evaluate the value of SPECT/CT in differentiating malignancy from benign spinal disease. Methods Fifty-three patients with foci of abnormally increased uptake in the spine detected by 99Tcm-MDP planar whole body bone scan subsequently underwent bone SPECT/CT. The final diagnosis was determined by pathological examination or clinical follow-up ( ≥6 months), which was applied to calculate the diagnostic efficacy of bone SPECT/CT. Results A total of 25 patients were confirmed to have bone malignancy. The diagnostic sensitivity, specificity, accuracy, false positive rate, false negative rate, positive predictive value and negative predictive value for 99Tcm-MDP bone SPECT/CT were 96.00% (24/25), 96.43% (27/28), 96.23% (51/53), 3.57% ( 1/28), 4.00% ( 1/25), 96.00% (24/25) and 96.43% (27/28), respectively. Conclusion 99Tcm-MDP bone SPECT/CT imaging provides good clinical value for the differential diagnosis of spinal diseases.

Background The parameter of corneal diameter in myopic eyes is widely used in clinic,but there are different points of view about the correlation of corneal horizontal diameter with other parameters of the myopic eye.Objective This study was to investigate the relevance of the other parameters to the corneal horizontal diameter(CHD)of myopia.Methods A total of 310 cases(310 eyes)of myopic patients aged 6-50 years old who visited Affiliated First Hospital of Zhengzhou University were collected for the study.Measuring items included gender,age,myopia diopter and corneal topography.The relationships between the CHD of the right eyes and seven factors including age,gender,degree of myopia,corneal curvature (CC),corneal astigmatism (CS),corneal central thickness(CCT),and anterior chamber depth (ACD)were analyzed by empower stats software.Results The distribution range of CHD was from 10.8 mm to 13.5 mm,with the average value (11.7±3.8)mm.There were significant differences in the CC and ACD between male and female patients by t test(t =-1.574,P＜0.001 ;t=-1.145,P =0.034).Through the smoothing curve fitting,the threshold effect and single factor and multiple regression analysis,the CHD was negative linear relationship with CC (β =-0.085,P =0.011).The ACD positive linear relationship with CHD was found (β=0.722,P＜0.001).And the CHD was not correlated with the degree of myopia,CS,CCT and gender(β =0.000,0.084,-0.001,0.105;all at P＞0.05).There was different inflection point in the curve relationship between male patients and female patients.Conclusions The CHD is linear negatively correlated with CC and line positively associated with ACD in 6-50 years old myopic patients.There is no relationship between CHD and gender,spherical equivalent degree,CS,CCT.There is curvilinear relationship with inflection point between CHD and age.

Acupuncture Therapy ; methods ; Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Muscle, Skeletal ; injuries

Objective To establish a stable animal model for sequentially dynamic and direct monitoring of the skin wound infection. Methods The mice with full-thickness skin incisions were replicated. After immediate subcutaneous suture,the mice were randomly divided into four groups,ie,Group A was inoculated with 50 μl sterile PBS solution),Groups B,C and D were inoculated with 50 μl suspension containing 1 × 106,1 × 108 and 1 × 1010 colony forming unit (CFU)/ml bioluminescent methicillin-resistant staphylococcus aureus (MRSA) respectively.Then,the diet behavior of each group was observed and the mean weight and mortality of each group were also recorded at different time points.The bioluminescent intensity of fluoresce in the wounds was recorded at different time points by using the charge-coupled device (CCD) based imaging system.Local wound tissues were incised at 24 hours after inoculation for HE staining so as to observe wound inflammatory reaction in each group.Wound healing time of each group was also recorded. Results ( 1 ) Average weight:Groups A and B showed unobvious changes in weight; Group C lightened until day 3 after inoculation and then recovered gradually to the preinoculation level at day 14; Group D lightened gradually until death.(2)Mortality:Groups A and B had no death; Group C had 10％ deaths at day 14; Group D had 100％ deaths.(3) Bioluminescent intensity of wounds:Groups A and B showed a gradual weakened luminescence since the day of inoculation and had almost complete disappearance at days 5 and 7 respectively; there was no sign of obvious increase or decrease in Group C from the day of inoculation till day 14 ; Group D had a gradual increase since the day of inoculation and the luminous area expanded until the death.(4) HE staining at 24 hours after inoculation:all the four groups showed inflammatory cell infiltration,especially in Groups C and D.(5) Wound healing time:wound healed at days 5 and 7 after inoculation in Groups A and B; the wounds showed no healing even at day 14 in the Group C,but the wounds length and area did not show obvious enlargement or diminishment ; the wounds extended gradually until the death in the Group D,since the day of inoculation. Conclusions The inoculation of 50 μl suspension with 1 × 108 CFU/ml bioluminescent MRSA to full-thickness skin incision rats allows direct,real-time dynamic and continuous detection of the occurrence and development of the wound infections.The infection model is easy to make and has stability and high repeatability.

Objective To explore the status,developing demands and directions of the education and training of military medical equipment.Methods The main problems of the PLA's medical equipment education and training were analyzed via being compared with those of foreign armies.According to the requirements on the campaign mode changing in the future war,the innovating mode under information-based military training,and the research and exploration of novel equipment and new technology,the developing directions in the future were ascertained.Results The interface was enhanced between new medical equipment development and educational training,and specifications were prepared for medical equipment application and maintenance.Conclutsion Exploring the pivot choke points and the developing directions will provide strategy guidance on the rapid and highly efficient elevation of military medical equipment supporting capacity.