Objective To analyze and evaluate the changes of quantity and function of TCBVα24+ Vβ11+ NKT in PBMC of the patients with PBC and its relationship with the occurrence of PBC. Methods Flow cytometry was utilized to count TCRVα24+ Vβ11+ NKT cells in PBMC in 60 cases of PBC and 60 cases of age-matched and gender-matched controls. NKT cells were activated and expanded by α-galcer and IL-2 in vitro. The percentages of positive NKT cells expressing IL-4 and IFN-γ were determined by ICS-FC. The levels of serum IL-4 and IFN-γ were tested by ELSIA. The numbers of cells secreting IFN-γ and IL-4 were detected by ELISpot. Results The ratios of NKT cells in PBC group were [0. 16(0. 11-0. 26) ]% before expansion and [0. 82 (0. 61-0. 89) ]% after expansion, significantly lower than control group [(0.33 (0.27-0.38) ]% and [27.40 (23.52-33.87) ]%, respectively, (Z=6.563, 7.707, P<0. 01 ). Seven days after expansion by α-galcer and IL-2, the expansion folds of NKT cells were 96. 05 (80.50-100.27) in PBC group and 134.65 (121.60-142. 13) in control group, respectively (Z =6.462,P < 0. 01 ). At the same time, the ratio of IFN-γ+ and TCRVα24+ Vβ11+ NKT detected by ICS-FC in PBC group was significantly higher than that in control group [ 38. 98 ( 36.73-42. 98 ) ]% vs [ 34. 56 ( 30. 12-36. 78 ) ] %, Z = 3. 158, P < 0. 05, while the ratio of IL-4+ and TCRVα24 + Vβ11+ NKT cells in PBC group was significantly lower than that in control group[0. 193(0. 179-0. 218) ]% vs [34. 36 (30. 93-38. 77) ]%,Z =6. 476, P <0. 01. The number of IFN-γ SFC detected by ELISpot were [410(380 ~500) ] SFC/1 × 106 PBMC in PBC group and [ 340(280 ~ 390)] SFC/1 × 106 PBMC in control group, respectively (Z = 4. 312, P <0. 05). The levels of serum IFN-γ in PBC group and control group were (67.21 ± 11.27) ng/L and (31.45 ± 8. 17) ng/L, respectively ( t = 27.25, P < 0. 01 ). The level of IFN-γ in PBC group was higher than that of control group. The number of IL-4 SFC were [73(60 ~ 100) ]SFC/1 × 106 PBMC in PBC group vs [245(230 -280) ] SFC/1 × 106 PBMC in control group, Z=5. 112,P <0. 01. The levels of serum IL-4 in PBC group and control group were (12.65 ±4. 17) μ/L and (28.31 ±6.31) μg/L, respectively (t =25.34,P < 0. 01 ). The level of IL-4 in PBC group was lower than that of control group. Conclusions The quantity of TCRVα2.4+ Vβ11+ NKT in PBC group is lower than that in control group. After in vitro activation, the capacity of expansion and producing IL-4 of NKT is decreased in PBC group, while the capacity of producing IFN-γ of NKT is increased in PBC group. The reduction of NKT cells and the immune dysfunction may be one of the important factors in the pathogenesis of PBC.

Objective To analyze the levels of Th17,CD4+ CDhigh25and the related cytokines (IL-17,IL-21,TGF-β,IL-10)in the plasma of patients with SLE.Methods The percentage of Th17 and CD4+ CDhigh25 Treg cells in peripheral blood were examined by flow cytometry.QRTPCR was used to detect the mRNA expression level of RORγt and Foxp3 of 56 SLE patients (26 patients are inactive and 30 patients are active) and 28 healthy controls.Plasma cytokines levels were measured by ELISA.Results The percentage of Th17 cells in patients with SLE was higher than that of healthy controls [ (3.44 ± 0.96) ％ vs (2.42 ± 0.52) ％,t =5.38,P =0.000 ].The percentage of CD4+ CDhigh25 Treg cells in patients with SLE was lower than that of healthy controls [ ( 1.32 ± 0.57 ) ％ vs ( 2.07 ± 0.67 ) ％,t =3.28,P =0.034].The expressions of RORγt mRNA was increased compared with healthy controls[ (0.219 ±0.063) vs (0.087 ± 0.045 ),t =6.41,P =0.000 ],but the expressions of Foxp3 mRNA was decreased compared with healthy controls [ (0.063 ±0.045) vs (0.128 ±0.056),t =5.28,P =0.000].Plasma levelsof IL-17,IL-21 and IL-10 in SLE patients were 122.4 ( 60.5 - 188.3 ),167.2 ( 128.7 - 871.4 ),51.3(20.9 - 123.7 ) ng/L,respectively,and in the control group were 27.1 ( 18.1 - 86.2 ),31.0 ( 31.0 -424.5),33.5( 16.4 - 54.1 ) ng/L,respectively,they were significantly different between the two groups (Z =3.83,4.54,1.87,all P ＜ 0.05 ).However,TGF-β levels were markedly decreased in SLE patients compared with controls [ 31.0 ( 31.0 - 168.6 ) ng/L vs 159.8 ( 63.4 - 389.7 ) ng/L,Z =4.87,P ＜0.05.IL-17 was positive correlation with Th17 cells,SLEDAI score,dsDNA antibody titers ( r =0.621,0.581,0.512,respectively,all P ＜ 0.05 ),but was negative correlation with C3 ( r =- 0.543,P ＜ 0.05 ) in SLE patients.Although the percentage of CD4 + CDhigh25 Treg cells were negatively correlated with SLEDAI score ( r =- 0.423,P ＜ 0.05 ),it was positively correlated with C3 ( r =0.511,P ＜ 0.05 ).Conclusion This study suggests that the imbalance of Th17 cells and CD4+ CDhigh25 Treg cells and the disorder of related cytokines are important in the pathogenesis of SLE patients.

Objective Study the value of clinical application of BaiJi and absorbable Gelatin in embolizing uterine myoma together with its effecty, side effect and complication.Methods 21 women with uterine myoma undergoing selective uterine arterial embolization by Seldinger's technique were studied. After retrograde transfemoral introduction of a 5 french catheter, the uterine arteries were successively catheterized. Bai Ji and absorbable Gelatin sponge particles were injected through free flow until devasculariztion. Results Uterine myoma's blood supply came from bilaterial uterine arteries demonstrated by angiography. All the supplying artering images disappeared after the embolization. 3～6 months follow up study showed: a marked reduction in the size of myomata by 38%～90%. Clinical symptoms were improved. There was one failure cas and then underwent uterotomy due to infection. Conclusions The short term effect of using Bai Ji and absorbable Gelatin for embolizing uterine myoma is clinically significant, while long term effects is still wating for research.

AIM: To investigate the effects of hypoxia-inducible factor-1 (HIF-1) on cardioprotection of hypoxic preconditioning (HPC) and its mechanisms. METHODS: In the model of hypoxia/reoxygenation (H/R) of cardiomyocytes from neonatal Spargue-Dawley rats, delayed cardioprotective effects of HPC on lethal H/R were observed. Whole cell extracts were prepared for determining activities of extracellular signal-regulated protein kinases (ERKs) and HIF-1? phosphorylation. RESULTS: HPC significantly promoted survival and membrane integrity of cardiomyocytes subjected to sustained H/R. SDS-PAGE mobility shift experiments showed increased phophorylation level of HIF-1? in hypoxic preconditioned cardiomyocytes. ERK1/2 were activated at 10 min after HPC and returned to the control level at 60 min. CONCLUSION: HPC protects neonatal cardiomyocytes from H/B injury. HIF-1? phosphorylation induced by ERKs might contribute to the cardioprotection of HPC.

Objective To investigate expression of β-site APP-cleaving enzymel(BACE1) and Aβ in brain of diabetes mellitus of Wistar rats,to study pathophysiological mechanism of Alzheimer disease from diabetic metabolic disorder. Methods Animal model of diabetes mellitus was established by streptozocin with intraperitoneal injection. Wistar rats were randomly divided into normal group (N), sham-operation group (S), 4 weeks diabetes mellitus model group (M4), 6 weeks diabetes mellitus model group (M6) and 8 weeks diabetes mellitus model group (M8). Behaviour was tested with Morris water maze and shuttle box test. Expression of Aβ was measured by enzyme linked immunosorbent assay and BACE1 by immunohistochemistry, enzyme linked immunosorbent assay, Western blotting and RT-PCR. The absorbance value was measured by imaging analysis. Results The electric times and latancy of memory and study were more increased in model group than that in N and S group but the times of escape more decreased(P<0.01). The expression of Aβ_(1-40) increased from (64.13±6.76)pg/mg in normal group to (86.43±7.03)pg/mg in model group by ELISA(P<0.001) and Aβ_(1-42) from (67.43±5.12 )pg/mg in normal group to (89.45±5.28) pg/mg (P<0.001) in model group. The expression of BACE1 increased from (116.46±8.10)pg/mg in normal group to (158.73±6.24)pg/mg in model group by ELISA and from 0.61±0.11 in normal group to 1.52±0.16 by Western blotting absorbance valule and from 1.62±0.26 in normal group to 3.61±0.32 by RT-PCR absorbance valule and from 0.81±0.21 in normal group to 2.01±0.36 by immunohistochemistry absorbance valule (P<0.001). The expression of BACE1 and Aβ in MT group was higher than that of in N and S group (P<0.01). The level of BACE1 and Aβ had positive correlation with cognitive impairment.Conclusion The expression of BACE1 and Aβ is increased in diabetes mellitus rats. Diabetes mellitus contributes to Alzheimer diseases that.

Objective To investigate the relationship between phosphatidylinositol 3-kinase catalytic subunit α(PIK3CA) expression and the incidence and different pathological grade of gastric cancer, and the mechanism thereof. Meth-ods The expressions of PIK3CA, serine/threonine protein kinase (pAkt) and cell proliferation associated nuclear antigen (ki-67) in gastric carcinoma and adjacent tissues and normal gastric mucosa were detected by immunohistochemical method. The relationship between expressions of PIK3CA, pAkt and ki-67 and tumorigenesis was analyzed. The expressions of PIK3CA, pAkt and ki-67 in different pathological conditions of gastric tissues were analyzed. The relationship between tu-mor pathologic classification and differentiation were analyzed too. Results There were significantly higher expressions of PIK3CA, pAkt and ki-67 in gastric cancer (P＜0.05), which were the highest in the poorly differentiated gastric adenocarci-noma (P＜0.05). There were a positive correlation between expressions of PIK3CA, pAkt and ki-67 and different pathologi-cal levels of gastric carcinoma and adjacent tissues and normal gastric tissues (P＜0.05). Conclusion PIK3CA may be the initiating factor of PI3K/Akt signaling pathway, which induced phosphorylation of Akt and activation PI3K/Akt signaling pathway, promoting the proliferation, invasion and metastasis of gastric adenocarcinoma.

AIM: To investigate the effects of human urotensin II (hUII) on ischemia/reperfusion (I/R) injury in isolated rat hearts. METHODS: In the ischemia/reperfusion (I/R) model of isolated perfused rat hearts,the effects of hUII pretreatment on cardiac function was monitored with cardiac function software of MFL Lab200. ATP,total calcium,and malondialdehyde (MDA) content in myocardium were detected. The coronary perfusion flow (CPF) and lactate dehydrogenase (LDH) activity in coronary effluent were measured during reperfusion. RESULTS: In the hUII pretreated group,the release of LDH from myocardium was lower [(78.3?18.1)U/L] than I/R group [(109.3?23.9) U/L,P

AIM: To investigate the effects of human urotensin Ⅱ (hUII) on in vivo mesenteric microcirculation in rats. METHODS: For recording of microcirculation images in the mesentery, the intestinal loop was mounted on the stage of an intravital microscope equipped with a TV camera. Video images of microcirculation were stored by a video cassette recorder. Temporal changes in internal diameter and microcirculatory velocity of microvesseles were measured by computer using the ImagePro software. The blood flow in intestinal wall was measured with PIMII laser Doppler perfusion Imager (Lisca Sweden). RESULTS: The internal diameters of arterioles and venules in control group were (21.4?2.3) ?m and (38.1?3.6) ?m,respectively. In UII group, the arterioles and venules contracted immediately after treated with UII and up to the peak at 1 min [(14.1?1.4) ?m and (22.2?5.2) ?m vs control,P0.05). The blood flow in intestinal wall increased 1 min after treated with UII and up to high peak at 5 min(6.4?1.1 perfusion unit vs control 4.2?0.9,P

AIM:To study whether ischemic preconditioning(IPC) has a protective effect against ischemia/reperfusion(I/R) injury in brain, and the possible relationship between IPC and the regulating function of microcirculation. METHODS: The I/R models were established both in I/R and IPC groups of Sprague-Dawley rats. Additional procedure was performed of short term cerebral ischemic preconditioning in IPC group 24 hours before I/R. Skull windows were performed through which microcirculation features were measured before ischemia, during ischemia, and reperfusion. Finally, brains were cut into slices and stained with red tetrazoline(TTC). RESULTS: Most TTC stained brains in I/R group presented irregular palely red areas which were few in IPC group. Compared with I/R group, IPC group presented relatively increase in accumulated length of capillaries, mean cerebral microcirculatory perfusion, and microcirculatory velocity in ischemic and reperfusion phase. There was no-reflow phenomenon in I/R group in reperfusion phase, which was substituted by the course of increasing reperfusion in IPC group. CONCLUSIONS:IPC could relieve the reduction of tissue perfusion during ischemia and the no-reflow phenomenon during reperfusion by improving the regulating function of microcirculation, which relatively promote the opening of capillaries and accelerating of microvascular flow, therefore protect brain from I/R injury.

OBJECTIVE: To detect potential mutation in a Chinese pedigree affected with congenital limb malformations. METHODS: Clinical data was collected. Genomic DNA was extracted from peripheral blood samples of family members. The zone of polarizing activity regulatory sequence (ZRS) were amplified by PCR and subjected to direct sequencing. RESULTS: Among the 13 individuals in this pedigree, there were 4 PPD patients, who were characterized by varying degrees of deformity. The female patients suffered triphalangeal thumb and preaxial polydactyly, while the male patients only had preaxial polydactyly. Only one patient had foot involvement. TA heterogeneous mutations was discovered in the ZRS (105C>G) in all patients, the same mutation was not detected in 2 healthy family members. CONCLUSION The inheritance pattern of PPD was autosomal dominant inheritance. There was a significant variability of symptoms among family patients. The heterozygous mutation of the ZRS (105C>G) probably underlie the disease.
Female ; Genetic Testing ; Hand Deformities, Congenital ; genetics ; Humans ; Limb Deformities, Congenital ; genetics ; Male ; Membrane Proteins ; genetics ; Pedigree ; Polydactyly ; genetics ; Thumb ; pathology