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Objective To select an optimal non-specific antigen blocking method by using immuno-infiltration assay so as to suit protein chip preparation. Methods Human papillomavirus type 16 L1 protein expressed by insect-baculovirus espressin system was incubated with skimmed milk powder, calf serum, bovine serum albumin (BSA) combinations of five kinds of methods to block the non-specific antigen. PBS was used as control. The effect of eliminating non-specific stain was detected by immuno-infiltration assay. Results After repeated tests, the results showed that the stability and repeatability of blocking effects were poor for the fixing up antigen first and then blocking method, and the blank control was prone to false positive. The infiltration rate of NC membrane would be affected by using skimmed milk powder as a blocking agent because the pore of NC membrane was easily plugged by milk powder particles. The use of calf serum as a blocking agent made it very difficult to determine the result because the calf serum absorbed by NC membrane produced the background; however, when 20g/L BSA was used to blocking before fixing up antibody, the results became satisfactory. Conclusion Fixing up antibody after blocking in immuno-infiltration assay showed that the blocking effect against non-specific antigen was satisfactory, stable and repeatable, indicating this method is a novel optimal blocking method compared with others.

Objective To investigate the prevalence of children dental fluorosis in drinking-water-borne endemic fluorosis areas in Liaoning province and to evaluate the disease prevention effects of water improvement measures.Methods Haicheng city,Linghai city,Faku county,Fumeng county,Liaoyang county and Jianping county six endemic fluorosis diseased counties (cities) of Liaoning province were included in the study in September 2011.In each county,3 water improvement villages were selected as survey points.In each surveyed village,children aged 8-12 were generally surveyed the prevalence of dental fluorosis.One tap water sample was collected in each surveyed village for determination of water fluorine.level.Results A total of 786 children aged 8-12 were examined; 110 of them were found with dental fluorosis; detection rate of dental fluorosis was 14.0％ ; no defect case of dental fluorosis was found,and dental fluorosis index was 0.27.Among the cases,very mild were 76 cases,and the detection rate was 9.7％; mild 33 cases,the detection rate was 4.2％; moderate 1 case,the detection rate was 0.1％; and no severe case.Four cases were in the eight-year-old group,accounting for 3.6％ of the total number of cases; 20 in the 9-year-old group,accounting for 18.2％ ; 26 in the 10-year-old group,accounting for 23.6％; 32 in the 11-year-old group,accounting for 29.1％; and 28 in the 12-year-old group,accounting for 25.5％.The differences of detection rates of dental fluorosis of children in each age groups were statistically significant (x2 =13.57,P ＜ 0.05).Water fluoride of the 18 water samples ranged from 0.34 to 2.03 mg/L; 77.8％ (14/18) of the water improvement areas reached the control standard for endemic fluorosis areas.Conclusion The prevalence of dental fluorosis in the areas with high fluoride in drinking water remains high,the measures of water improvement should be strengthened.

BACKGROUND:The osmotic tissue expander is a self-fil ing device consisting of an osmotic active hydrogel which is made of vinylpyrrolidone and mehtylmethacrylate. It can absorb body fluids and swel up gradual y after embedded.
OBJECTIVE:To explore the short-term and long-term regular patterns as wel as histocompatibility of the osmotic tissue expander in vivo.
METHODS:A self-control design was carried out in Wistar rats by embedding the osmotic tissue expander and high-density polyethylene into each side of their spinal column subcutaneously. Wound healing, tissue expansion and inflammatory reaction were detected and compared at different periods after operation.
RESULTS AND CONCLUSION:Al the wounds got primary healing. The device expanded fastest at week 1 after the implantation. After being enlarged to about nine times that of the initial size at week 4, the expander slowed down its swel ing. It reached its ultimate volume at week 12 which was about 10 times as big as that of the initial one. Then it remained almost the same size until the end of our design. Pathological sections showed that the inflammatory reaction of osmotic-tissue-expander-group had no significant difference from that of the control group (P>0.05). These findings suggest that osmotic tissue expander has a slow-lasting swel ing ability and good histocompatibility.

Objective To investigate the role of heparin binding epidermal growth factor-like growth factor(HB-EGF)in active psoriasis vulgaris.Methods HB-EGF mRNA and protein were detected by hy-bridization in situ and immunohistochemistry in normal skin tissues,lesional and non-lesional psoriatic skin of progressive stage.Results In normal skin tissues,the stain of HB-EGF mRNA and protein was located in the basal layer of epidermis(100.00%)and there was only a little expression in the suprabasal layers(16.67%).Focal overexpression was found in the suprabasal layers of non-lesional and peri-lesional psoriat-ic skin(88.00%,80.00%respectively);however,there was no HB-EGF mRNA protein expression in the superabasal layers of the central part of psoriatic lesions(0),and nearly no expression in the basal layer(4.00%).Conclusion HB-EGF may play an important role in the pathogenesis of early psoriasis.

Objective To observe the effect of electroacupuncture on back three-acupoints ( namely bilateral Dazhu, Fengmen, Feishu) on the protein and mRNA expression of transforming growth factor-beta 1 (TGF-β1) and Smad3 in asthmatic rat airway remodeling model, and to explore its therapeutic efficacy and molecular mechanism. Methods Rat asthma model was established by inhalation of ovalbumin. After sensitization for 6 weeks, rats were killed. And then the airway morphological parameters of rats were measured by image analysis. The protein and mRNA expression of TGF-β1 and Smad3 in lung tissues were detected by immunohistochemistry and real-time quantitative polymerase chain reaction (qRT-PCR) respectively. Results Compared with the blank group, the standardized values of muscle cross-sectional area including airway smooth muscle area (WAm) /perimeter of the basement membrane (Pbm), and bronchial inner wall area (WAi)/Pbm were increased in the model group. The protein and mRNA expression levels of TGF-β1 and Smad3 were also increased in the model group. In electroacupuncture group, the above observation indexes were decreased, and the differences were statistically significant ( P<0.05 compared with the model group). Conclusion The experimental results indicated that back three-acupoint electroacupuncture has an effect on remodeling airway, and one of the mechanisms is probably associated with the regulation of TGF-β1/Smad3 signaling pathway.

Aim To investigate the protective effect of cinnamic aldehyde ( CA ) on hormone-induced osteo-clasts proliferation and bone resorption in vitro and its molecular mechanisms. Methods RAW264. 7 cells induced into osteoclast were treated with RANKL and M-CSF and then were divided into control group, dexa-methasone ( DEX ) group and different doses of CA (11. 6, 23. 2, 46. 4 μg·L-1 ) groups. OCs were ob-served after tartrate resistant acid phosphatase( TRAP) staining. The cell proliferation was determined by MTT assay at different time points. The expression levels of TRACP5 b in cell cultured supernatants were measured by ELISA. RT-PCR technique was applied to examine the transcriptional levels of RANK and NFATc1 . Re-sults In MTT assay, the proliferation of osteoclasts stimulated by dexamethasone was promoted seriously compared with negative control group ( P < 0. 05 ) . Meanwhile, DEX could strengthen the content of TRACP5 b and up-regulate the expressions of RANK and NFATc1 mRNA. After administration of CA, the proliferation was inhibited, while the enhanced expres-sion of TRAP5b was reversed,and the over-expressions of RANK and NFATc1 mRNA were obviously down-regulated in a time-and-dose-dependent manner ( P <0. 05 ) . Conclusion The results suggest that CA in-hibits proliferation and bone resorption of osteoclast in-duced by DEX, which may be mediated by down-regu-lation of RANK and NFATc1 mRNA.

Objective To investigate clinical manifestations and clinical diagnosis and treatment and pathological and immunohistochemical features in gastrointestinal stromal tumors. Methods The clinical data of fifty-two cases with gastrointestinal stromal tumors were collected, whose clinical diagnosis and treat-ment and pathological features were retrospectively analyzed from January 1995 to December 2007. Results All patients received operation therapy, only forty-five cases with complete surgical resection. The immu-nohistochemical staining showed that the cases with CD117 positive accounted for 100% (52/52) and CD34 positive accounted for 88.5% (46/52). Conclusions Surgery was necessary for all patients, especially complete surgical resection. Gastrointestinal stromal tumors were poor in preoperative diagnosis, which diag-nosis was based on the immunohistochemical staining of the tumor tissue. CD117 and CD34 tumor markers may help to diagnose gastrointestinal stromal tumors.

Objective To investigate the correlation between apparent diffusion coefficient (ADC)value of MR diffusion weighted imaging (DWI)and Ki-67 expression in rectal cancer.Methods The data of 47 patients with rectal cancer confirmed by pathology were retrospectively analyzed.Pelvic MRI,rectum DWI and enhanced examination were performed preoperatively.According to the Ki-67 index,the patients were divided into A-D four Groups:Group A with Ki-67 <10% (-),Group B with Ki-67 10%-25%(+),Group C with Ki-67 26%-50%(++);and Group D Ki-67 > 50%(+++).ADC values were calculated in each group to analyze the correlation with the Ki-67 expression levels.Results The mean ADC value were (1.280 ± 0.200 )× 10 -3 mm2/s in Group A (13 cases),(1.044±0.046)×10 -3 mm2/s in Group B (14 cases),(0.987 ±0.256)×10 -3 mm2/s in Group C (9 cases), and (0.85 1±0.099)×10 -3 mm2/s in Group D (1 1 cases).The mean ADC values were significantly different in the different groups with different levels of Ki-67 expression (P =0.001).ADC value and Ki-67 expression were moderate negatively correlated (r = -0.577, P=0.000).Conclusion DWI and ADC measurement can evaluate indirectly the cell proliferation and prognosis of rectal tumor to some extent.

Objective To evaluate the role of calcium/calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) in the up-regulation of expression of Cav3.2 T-type calcium channels in spinal cord in a rat model of neuropathic pain (NP).Methods Forty-eight male Sprague-Dawley rats,aged 3 months,weighing 220-250 g,were randomly divided into 6 groups (n =8 each)∶ sham operation group (group S),group NP,dimethyl sulfoxide group (group D) and different concentrations of a specific CaMK Ⅱ inhibitor KN93 groups (groups K1-3).NP was produced by chronic compression of dorsal root ganglion.The rats in groups D and K1-3 received a single intrathecal injection of dimethyl sulfoxide and KN93 15,30,60 nmol/L (10 μl),respectively,on 5th day after NP.Paw withdrawal threshold to von Frey filament stimulation (MWT) and paw withdrawal latency to thermal nociceptive stimulus (TWL) were measured before NP,before intrathecal injection on 5th day after NP,and at 30 and 60 min and 3,6 and 8 h after intrathecal injection on 5th day after NP (T1-7).The rats were sacrificed after the measurement of pain threshold at T7 and their lumbar enlargements were removed to detect the expression of Cav3.2 mRNA and protein using Western blot and RT-PCR.Results Compared with group S,MWT was significantly decreased,TWL was shortened and the expression of Cav3.2 mRNA and protein was up-regulated in NP,D and K1-3 groups (P ＜ 0.05).Compared with NP group,MWT was significantly increased,TWL was prolonged and the expression of Cav3.2 mRNA and protein was down-regulated in a concentration-dependent manner in K1-3 groups (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group D (P ＞ 0.05).Conclusion CaMK Ⅱ is involved in the development and maintenance of chronic NP by up-regulating the expression of Cav3.2 T-type calcium channels in rat spinal cord.