Objective To investigate the roles of phosphorylated glycogen synthase kinase 3β (pGSK3β) and phosphorylated signal transducer and activator of transcription 3 (pSTAT3) in hypoxic preconditioning-induced neuroprotection against ischemic brain injury in rats.Methods Sixty SD rats were randomly divided into a sham operation group,a cerebral ischemia group,and a hypoxia preconditioning group (n =20 in each group).A model of middle cerebral artery occlusion (MCAO) was induced by the modified suture method.Before the preparation of MCAO model,the rats in the hypoxia preconditioning group were put into a hypobaric oxygen chamber at a simulated altitude of 5 000 m (pressure:0.53 × 105 kPa;partial pressure of oxygen:81 mmHg;1 mmHg =0.133 kPa),3 h a day for 5 days.At 24 h after MCAO modeling,the rats were subjected to neurobehavioral score (n =6) and cerebral infarction volume measurement (n =6).Immunohistochemical staining was used to detect the expression levels of neuronal nuclei (NeuN) and pGSK3β (Ser9) (n=7).Western blot was used to detect the expression levels of pGSK3 β (Ser9) and pSTAT3 (Tyr705) in the ischemic cortex (n =7).Results The neurological deficit score (1.833 ±0.408 vs.2.667 ± 0.516;t =3.101,P=0.011) and cerebral infarction volume (18.137％ ± 0.801％ vs.24.125％ ± 0.694％;t =13.840,P＜ 0.001) in the hypoxia preconditioning group were significantly lower or smaller than those in the cerebral ischemia group.Immunohistochemical staining showed that the numbers of NeuN positive cells in the cerebral ischemia group and the hypoxia preconditioning group were significantly less than that in the sham operation group (48.000 ± 1.414/high power field [HPF],124.833 ± 3.061/HPF,and 213.500 ± 2.429/HPF;F =7 150.550,P ＜ 0.001),the hypoxia preconditioning group was significantly more than the ischemia group (P ＜0.001);the numbers of pSTAT3 positive cells in the cerebral ischemia group and the hypoxia preconditioning group were significantly higher than that in the sham operation group (57.667 ± 1.366/HPF,29.167 ± 1.941/HPF and 3.500 ± 1.049/HPF;F =1 962.649,P ＜0.001),and the hypoxia preconditioning group was significantly less than the ischemia group (P ＜0.001).Western blot analysis showed that the expression levels of ischemic cortical pGSK3β and pSTAT3 in the cerebral ischemia group and the hypoxia preconditioning group were significantly higher than those of the sham operation group (pGSK3 β:2.336 ± 0.102,0.876 ± 0.196 and 0.440 ± 0.012;F =1 610.826,P ＜ 0.001;pSTAT3:8.368± 0.230,4.883± 0.123 and 0.595± 0.138;F=4018.051,P＜0.001),the hypoxia preconditioning group were significantly lower than the ischemia group (all P ＜0.001).Conclusions Hypoxia preconditioning has neuroprotective effect for ischemic brain injury in rats.It may be associated with the down-regulation of the expressions of pGSK3 and pSTAT3.

Objective To investigate the relationship between angiotensinogen(AGT)gene M235T polymorphism and the onset of chronic heart failure(CHF).Methods Polymerase chain reaction combined with restrictive fragment length polymorphism was used to detect the M235T variants of AGT gene in 111 CHF cases,accompanied with 110 healthy controls.Results Three kinds of AGT genotypes were detected,which were MM、MT and TT genotype separately.The frequencis of TT genotpye were higher in CHF cases than in healthy controls,and there was statistical significance in old aged group(P =0.042).The odds ratio(OR)with CHF of TT genotype was 3.367 compared with non-TT genotypes in old aged group.Conclusion The M235T polymorphism of AGT gene is associated with the occurrence of CHF in old aged group and TT genotype might be one of the genetic risk factors of CHF in such place of old aged population.

Objective To explore the expression of matrix metalloproteinase-14(MMP-14)protein in the human stomach carcinoma tissues and its correlation with carcinoma invasiveness and metastasis.Methods The MMP-14 protein expression was detected by immunohistochemistry in 59 cases of stomach carcinoma tissues (observation group)and 20 cases of normal stomach tissues (control group,the adjacent normal tissues from the tumor margin of 5 cm confirmed by pathology),and its correlation with the clinically pathological parameters was analyzed.The expression characteristics of MMP-14 among various TNM stages of stom-ach carcinoma were also analyzed.Results The positive rate of MMP-14 expression was 50.85%(30/59)in the observation group and 5.00% (1/20)in the control group,the positive rate of the observation group was significantly higher than that of the control group (P <0.01);the expression level of MMP-14 was correlated with the differentiation degree,regional lymph node metastasis degree,invasion depth,lymphatic invasion and TNM stage,which showing the statistical difference(P < 0.01);the expression of MMP-14 protein was up-regulated and showed the transferring trend from cytoplasm to cellular membrane along with the progres-sion of TNM stage.Conclusion The overexpression of MMP-14 protein exists in stomach carcinoma tissues,which contributing to the invasion and metastasis of stomach carcinoma cells.

Objective To investigate the effects of cyclooxygenase-2 inhibitor celecoxib on proliferation , invasion and migration of human pancreatic cancer cell line PANC-1 and then determine the optimal concentration of cele-coxib and the most suitable application time .Methods Human pancreatic cancer cell line PANC-1 was treated with diverse concentrations of celecoxib (20,60,100 μmol/L) for different durations (24,48,72 h).Cell prolifer-ation, invasion and migration capabilities were measured by MTT colorimetry , Transwell invasion assay , and scratch assay respectively .Results The proliferation capability of PANC-1 cell was reduced by celecoxib in a con-centration-and time-dependent manner ( P <0.05 ) .In addition , the invasion and migration capabilities were decreased by celecoxib in a concentration-dependent manner(P<0.01).Conclusions Celecoxib attenuates the proliferation of human pancreatic cancer cell line PANC-1 in a concentration-and time-dependent manner .Cele-coxib attenuates the invasion and migration in a concentration-dependent manner .

Objective: To observe the clinical effects of a modified Tuina method in the treatment of childhood diarrhea. Methods: Thirty children with diarrhea of spleen deficiency were treated by Four-Step Tuina(treatment group); and control group was treated with routine Tuina methods. The scale of children diarrhea was used as an objective index for therapeutic effect evaluation. Results: After treatment, the scale was 1.4±1.92 in treatment group and 3.3±3.45 in control group; the statistical analysis showed that there was a difference between the scales; the treatment group had better effects than control group, in another word, the Four-Step Tuina was quite effective for children diarrhea. Conclusion: Tuina has positive effects on children diarrhea of spleen deficiency, which provides evidences for the standardization of clinical therapeutic effects.

Thirty-three infants with muscular torticollis were treated by Tuina techniques, and the size of the hardened sternocleidomastoid muscle in the affected side was B-ultrasonographically observed before and after treatment. Results: 27 cases were cured, 5 cases got effectiveness, 1 case failed and the total effective rate was 97.0%; B-ultrasonography, in the palpable cord-like or ovary-like lumps,showed low echo in 28 cases, moderate echo in 2 cases and high echo in 3 cases; there were significant differences in the length, thickness and width of the lumps between before and after treatment, P＜0.01. It is indicated that Tuina is quite effective against infantile muscular torticollis; that ultrasonography can be adopted as an objective item for the evaluation of the clinical efficacy of Tuina in the treatment of infantile muscular torticollis, and it is a simple, convenient and inexpensive method and should be extensively popularized in clinical practice.

Objective To investigate the anti-tumor effect and mechanism of curcumin in pancreatic cancer (PC). Methods Smad4 and Jab1 expressions were detected by immunohistochemistry in tumor tissues and pericarcinomatous tis?sue from 35 PC cases, and the correlation of Smad4 and Jab1 were analyzed based on the percentage of positive staining in?tissues from 21 random selected PC cases. The effect of curcumin on expressions of tumor suppressors p53, Smad4 and cell cycle inhibitor p27 were examined by Western Blotting after human pancreatic cancer cell line PANC-1 were divided into PANC-1 control group (no treatments were given) and PANC-1 curcumin group (treated with cell culture medium containing 10μmol/L curcumin). The effect of curcumin on expressions of combination of β-TrCP1 and Smad4 was examined by Co-Immunoprecipitation after human embryonic kidney cell line 293T were divided into 293T control group (no treatments were given), 293T curcumin group (treated with cell culture medium containing 10μmol/L curcumin) and 293T Jab1 group (trans?fected by HA-Jab1 plasmid). Results Compared with expressions in pericarcinomatous tissues, Smad4 was down regulated while the expression of Jab1 was upregulated in PC tissues (P<0.01), and the expression of Smad4 was negatively correlated with the expression of Jab1 (n=21, r=-0.71, P=0.007). After treated with curcumin, the protein expression of p53, Smad4 and p27 was increased in PANC1 cell, and the protein expression of the combination ofβ-TrCP1 and Smad4 was decreased in 293T cell (P<0.05). After transfected by HA-Jab1 plasmid, the protein expression of the combination ofβ-TrCP1 and Smad4 was increased in 293T cell (P<0.05). Conclusion Curcumin may have suppression effect of PC through increasing the protein expression of p53, Smad4 and p27, and the mechanism of Smad4 upregulation may be related with the inhibition of Smad4 ubiquitination process, while Jab1 may be also involved in Smad4 degradation through ubiquitination.

Objective To investigate the accuracy for absolute quantification of brain creatine (Cr) concentration using long time echo (TE) point resolved spectroscopy (PRESS) sequence performed with an external standard and postprocessed with the linear combination of model spectra (LCModel).Methods Ten swine (3.1±0.6 kg) and an external standard phantom containing detectable compounds of known concentration were investigated in this study by using 1.5 T GE Signa scanner and a standard head coil. The single-voxel proton magnetic resonance spectroscopy (1H-MRS) data were acquired from the two ROIs (2 cm×2 cm×2 cm) placed in swine brain and external standard solution using PRESS sequence with TE 135 ms,TR 1500 ms,and 128 scan averages.The in vivo quantification of Cr was accomplished by LCModel.After 1H-MRS examination,each animal was sacrificed immediately.In vitro Cr concentration was analyzed by high performance liquid chromatography (HPLC).Results In the 1H-MRS group,the Cr concentration was (9.37±0.14)mmol/kg.In the HPLC group,the Cr concentration was (8.91±0.13)mmol/kg.Good agreement was obtained between these two methods (t=9.038,P=0.491).Conclusion The long echo time PRESS sequence performed with an external standard and processed with LCModel iS proven to be an accurate technique to detect the in vivo brain Cr concentration.

Objective To evaluate the absolute quantification of brain metabolites concentrations using external standard MRS in acute hypoxia ischemia encephalopathy (HIE) piglet model. Method Eight 7-day-old healthy piglets were subjected to insult of hypoxia ischemia (HI). The animals and an external standard phantom containing detectable metabolites of known concentrations were studied on a 1.5 T GE Signa scanner. The single-voxel proton magnetic resonance spectroscopy (1H-MRS) data were processed using LCModel software, and the quantification of N-acetylaspartate ( NAA), creatine (Cr) and lactate (Lac) were accomplished. Multivariate analysis of variance was performed to compare the NAA, Cr, Lac concentration differences in the brains of piglets pre- and post-HI (0h). In addition, the dynamic changes of brain metabolites concentrations of 2 HIE piglets were observed at the time points of 0 h and 2 h. Results One piglet was excluded because it was over anesthetized to death. Seven piglets' data were analyzed. The concentrations of NAA pre- and post-HI were ( 6. 86 ± 0. 49 ) mmol/kg and ( 5.73 ± 0. 88 ) mmol/kg respectively, they were ( 4. 65 ± 0. 73 ) mmol/kg and ( 4. 40 ± 0. 80 ) mmol/kg for Cr; and were 0. 00 mmol/kg and (0. 43 ± 0. 39) mmol/kg for Lac. After HI, decreased NAA concentration immediately was observed, and it was of statistical significance ( F = 8. 608, P = 0. 013 ). The concentration of Cr was insignificantly decreased ( F = 0. 379, P = 0. 550). The concentration of Lac was increased, and the difference was of statistical significance ( F = 8. 600 ,P = 0. 013 ). Dynamic observation showed a Lac peak immediately after HI and it decreased after 2 h post-HI. Conclusions External standard MRS using LCModel has great value in the quantitative analysis of brain metabolites. The changes of NAA and Lac concentrations are sensitive to reflect the early metabolic change of acute HIE.

Objective To study the immediate conservative breast surgical reconstruction by trans-ferring the same lateral latissimus dorsal myocutaneous flap (LDM) for the treatment of stage Ⅱ and stage Ⅲ breast cancer, combined with neoadjuvant chemotherapy. Methods Breast cancer patients in stage Ⅱ and Ⅲ with pathological diagnosis by core needle biopsy, had undergone immediate conservative breast reconstruction surgery with transferring the latissimus dorsal myocutaneous (LDM). We scored the reconstructed breast twice at one month after operation and completion of radiation. The final score was decided using the mean value of the twice evaluating score. Kaplan-Meier survival was used to analyze the survival of 18 cases with traditional mastectomy. Results The tumor size ranged from 30 mm to 55 mm before neoadjuvant chemotherapy, and ranged from 25mm to 45 mm after neoadjuvant chemotherapy. The median weight of the tumor specimens in breast conservative operation was 140 g (90 g to 220 g). A-mong the 18 patients, 16 cases had more than 3 scores with satisfied cosmetics (93.33 % ). Dorsal subcu-taneous seroma in donation area was observed in 5 of 18 patients (27.78 %) and dorsal incision dehiscence was observed in 2 of 18 patients (11.11%). None of the patients developed recurrence except that 1 pa-tient (5.55 %) had distance metastasis (bone metastasis} in a mean 22-months follow-up. The Kaplan-Meier survival curve in breast conservation reconstruction patients had no different from that of traditional mastectomy patients at the same time. Conclusions Immediate conservative breast surgical reconstruction by using the same lateral latissimus dorsal myocutaneous flap is an effective method for stage Ⅱ and stage Ⅲ breast cancer patients combined with neoadjuvant chemotherapy, which can increase satisfactory breast so that cosmetic outcome can be reached.