Objective To clone human survivin gene and express survivin-an inhibitor of apoptosis proteins in Pichia pastoris eukaryotic expression system. Methods Full length survivin gene was amplified by PCR using survivin specific primers. The verified survivin gene by sequencing was subcloned into pPic9k. The recombinant plasmid was linearized by restriction enzyme cutting. The linear survivin gene was introduced into GS115/ His-cells by electroporation. The transformants were transferred onto YPD plates that contained different concentrations of G418 for screening the positive clones. The integrated survivin gene in positive clones was confirmed by PCR. Selected transformants were cultured in BMMY medium with 1 % methanol for inducing the expression of survivin protein. The expressed survivin protein was analyzed by ELISA. Results The cloned human survivin sequence was the same as that in the GeneBank. The recombinant pPic9k-survivin was constructed in Picha pastoris eukaryotic expression vector. After the linear digestion, the recombinant vector was introduced into GS115/ His-cells, the 6 positive clones against G418(4 mg/mL) were obtained and confirmed by PCR; the highest survivin protein was expressed when expressed for two days in 1 % methanol BMMY medium. Conclusions Improved survivin protein yield may be reached by modifying the experimental conditions. This will help to further study the biological functions of survivin and its roles in tumor developments.

Objective To quantify iron deposition in Alzheimer’s disease (AD),amnestic mild cognitive impairment (aMCI), and normal old people using susceptibility weighted imaging (SWI).Methods Sixty participants (22 aMCI,20 AD,18 normal con-trols)underwent conventional magnetic resonance imaging sequences and SWI using axial/oblique coronal sequences.Phase images were used to calculate bilateral iron deposition in 1 8 regions of interest (ROI)of every participant.The radian angle values were cal-culated and compared between the three groups.Results The radian angle value difference was significant between the aMCI and control groups in the left (L)-hippocampus,L-head of the caudate nucleus,R/L-lenticular nucleus ROIs.The radian angle value difference was significant between the AD and aMCI groups in the R/L-cerebellar hemisphere,R/L-hippocampus,R-red nucleus, R/L-thalamus and splenium of corpus callosum ROIs.Pearson correlation coefficients of the mini-mental state examination (MMSE)scores and many ROIs were significant.Conclusion Iron depositions in the hippocampus,head of the caudate nucleus,len-ticular nucleus and thalamus are significantly different between individuals with aMCI,AD and controls.Using SWI to quantify the iron deposition is a useful tool in detecting aMCI and AD.

ABSTRACT Recent research suggests that the ?-amyloid peptide (A?) is central to the pathophysi-ology of Alzheimers disease (AD). Amyloid plaques, primarily composed of A?, have significant neurotoxic effects. So it is therapeutic strategies for AD to lower the production of A? in the brain. A? is a product of catabolism of amyloid precursor protein ( APP) by ?-secretase and y- secretase. BACE ( ?-secretase) exhibits all the properties of the ?-secretase, and as the key rate-limiting enzyme that initiates the formation of A?, BACE is an attractive drug target for AD. The recent research of ?-secretase is reviewed in this paper.

AIM: To apply high throughput screening methods to research Xiaoxuming Decoction's effective components combination and its action mechanism. METHODS: We studied the effects of 240 sequential components of Xiaoxuming Decoction (L1-L120,A1-A120) on anti-A? neurotoxicity、anti-H_2O_2 damage、anti-glutamic acid damage and ?-secretase activity. RESULTS: We evaluated these results of screening comprehensively,and found several components of Xiaoxuming Decoction (L1-L40,A30-A60,A100-A120) had the potential effects of listed before.So the combination of these components was regarded as the effective components combination of Xiaoxuming Decoction for anti-Alzheimer's disease. CONCLUSION: Xiaoxuming Decoction had effective protection against Alzheimer's disease through multi-component and multi-target.High throughput screening methods will push the development of Chinese traditional compound prescription greatly.

AIM: To establish the model of aging rats and observe the effect of the group of effective components of Xiaoxuming Decoction(XXM)on aging rats. METHODS: D-galactose and rotenone were used to establish aging animal model.The ability of learning and memory were measured by using Morris water maze test.Then, the content of GSH and MDA,the activity of SOD,the activity of AchE and ChAT in rats'brain were measured. (RESULTS): D-galactose could induce aging like indicators in rats:decrease the content of GSH and the activity of SOD,and increase the content of MDA,but have no effect on AchE and ChAT.It showed that GEC could improve the ability of learning and memory and ameliorate biochemistry function in rats with aging induced by D-galactose. CONCLUSION: These results suggest that GEC of XXM could improve the responses of aging rats and these effects may be related to its ability of antioxidation.

Objective To investigate the effect of atorvastatin on cytochrome c (CytC) expression and neuronal apoptosis after intracerebral hemorrhage in rots.Methods A total of 108 male Sprague-Dawley rats were randomly allocated into 3 groups:sham operation group,saline control group,and atorvastatin group (n =36 each group).All the groups were redivided into 6 h,12 h,day 1,3,5 and 7 time points (n =6 at each time point).An intracerebral hemorrhage model was induced by using a modified two-step injection method.After modeling,atorvastatin was used for gavages (20 rng/kg,once a day) in the atorvastatin group.The saline control group was given the same volume of saline.Behavior evaluation was used for neurological score.TUNEL staining was used to detect apoptosis in perihematoma tissue.Immunohistochemical method was used to detect the CytC expression in perihematoma tissue.Results Behavior evaluation showed that the neurological scores decreased gradually with the passage of time in the atorvastatin group and the saline control group.There were no significant differences at 6 h,12 h,day 1 and day 3,but the neurological scores in the atorvastatin group were significantly lower than those in thc saline control group at day 5 (0.50 ± 0.55 vs.1.50 ± 0.55; t =3.162,P =0.010) and day 7 (1.00 ±0.63; t =2.712,P =0.022).TUNEL staining showed that the numbers of apoptotic cells increased first and then decreased in the saline control group and the atorvastatin group.They reached the peak at 1 hour after modeling.There were significant differences in the number of apoptotic cells in each group in perihematoma tissue at the same time point (all P =0.000),and the significance in the saline control group was more than that in the sham operation group and the atorvastatin group (all P ＜0.05),but at day 7,there was no significant difference in the number of apoptotic cells between the atorvastatin group and the sham operation group (12.69 ± 3.35 vs.9.33 ± 2.07; P =0.148).Immunohistochemical method showed that the numbers of CytC positive cells increased first and then decreased in the saline control group and the atorvastatin group,reached the peak at 12 h after modeling in te saline control group (68.19 ± 11.93) and at 1 d in the atorvastatin group (35.64 ± 9.12).There were significant differences in the numbers of CytC positive cells in perihematoma tissue at the same time point in each group (P =0.000).The numbers of CytC positive cells in the saline control group was significantly more than that in the sham operation group and the atorvastatin group (all P ＜0.05),but there was no significant difference in the numbers of CytC positive cells between the atorvastatin statin group and the sham group at day 7 (16.08 ± 3.80 vs.13.67 ± 2.94; P =0.349).Conelusions Atorvastatin may inhibit the release of CytC of nerve cells in perihematoma tissue after intracerebral hemorrhage,and thus reduce CytC-mediated apoptosis and neurological deficit after intracerebral hemorrhage.

Objective:Until recently, no consensus has been reached about the treatment of primary tumor in patients with meta-static breast cancer, and whether or not to excise it has not yet reached agreement. This study aimed to evaluate the value of surgical and nonsurgical treatment of primary tumor by analyzing the clinical data of patients with metastatic breast cancer. Methods:This re-view includes the data of 120 metastatic breast cancer patients. Their clinical data in Xiangyang Central Hospital (Hubei province) from January 2005 to December 2012 were collected. All cases were divided into surgical and nonsurgical groups, and the overall survival and symptomatic local progression rates were analyzed. Results:The 120 patients had a median follow-up of 52 months (range=10-92 months). A total of 55 cases were in the surgical group, 30 of whom had surgery before the metastatic diagnosis, and 65 cases were in the nonsurgical group. No significant differences were observed regarding the tumor classification, lymph-node classification, and meta-static site of the tumor in the two groups. Patients in the surgical group experienced longer overall survival (49 months vs. 33 months, P=0.016) and lower rates of symptomatic local progression (14.5%vs. 46.2%, P<0.001). Conclusion:This study demonstrated that the overall survival and symptomatic local control in the surgical group were better than those in the nonsurgical group. However, this hy-pothesis remains to be proved by multicenter clinical trials.

Although digital subtraction angiography (DSA) has been used as the gold standard for the diagnosis of intracranial aneurysms, its clinical use is somewhat limited by its invasiveness, high medical cost and potential risk of nephrotoxicity. For the past three decades, the magnetic resonance angiography (MRA) techniques have been developed rapidly. As a non-invasive technique with high resolution ability, MRA can replace DSA for the diagnosis of intracranial aneurysms in most clinical situations. The time-of-flight MRA (TOF-MRA) carries a diagnostic sensitivity of up to 98.2%-98.7%for tiny intracranial aneurysms (<5 mm), while the diagnostic sensitivity and specificity of contrast enhanced MRA (CE- MRA) for intracranial aneurysms are as high as 95% - 100% and 73% - 100% respectively. The diagnostic specificity of phase-contrast MRA (PC-MRA) for intracranial aneurysms (> 5 mm) reaches as high as 100%. All the above mentioned MRA techniques can clearly display the intracranial aneurysms although their imaging characteristics and clinical applications are different from each other. This paper aims to make a brief review concerning the principles, clinical applications and recent progress of some MRA techniques.

Objective To find an easy and proper way to differentiate metastatic breast cancer from second primary breast cancer by analyzing the histopathological characteristics of 40 patients with bilateral breast cancer.Methods According to the Chaudary histological criteria,the histopathological and biological characteristics of 16 cases of synchronous bilateral breast cancer and 24 cases of metachronous bilateral breast cancer were evaluated.The histopathological and biological characteristics included Nottingham histological grade,immunohistochemistry (IHC) results of the estrogen receptor,progesterone receptor and expression of Her-2.Results The average age of the 40 patients at first diagnosis was 41 years old (range,26-68 years old).The average time interval between first and second diagnosis of tumors was 34 months (range,7-209 months) in metachronous cancer.The concordant histopathological type was found in 93.8 ％ (15/16) of synchronous cancer patients and 58.3 ％ (14/24) of metachronous cancer patients (P =0.02).The concordance rates of tumor stage was 68.8 ％ (11/16) in synchronous cancer patients,while it was 25.0 ％ (6/24) in metachronous cancer patients (P =0.03).For progesterone receptor status,the concordance rates were 87.5 ％ (14/16) and 54.2 ％ (13/24) in synchronous and metachronous cancer patients respectively (P =0.03).There was no statistically significant difference in terms of estrogen receptor status and Her-2 expression (P ＞ 0.05).Conclusions Without considering the limitation of Chaudary criteria and the number of patients,it seems to be an easy and proper way to differentiate metastatic cancer from second primary cancer in the patients with bilateral breast cancer by combining the histopathological type,tumor stage and progesterone receptor status.The synchronous cancer is closer to same clonal lesion (metastatic lesion).

Objective To investigate the psychological status and related factors in patients with mainte-nance hemodialysis in Huzhou city.Methods 376 cases of maintenance hemodialysis were selected as the research subjects in Huzhou First People's Hospital.The anxiety and depression status in maintenance hemodialysis patients were investigated by issuing questionnaires.To study the relationship between anxiety and depression,gender,age, social support,nutritional status,complications,and in maintenance hemodialysis patients.At the same time,the quality of life was evaluated by SF -36 scale.Results Issuing 376 questionnaires,a total of 373 points,the effective recovery rate was 99.20%.In 373 patients,there were 182 patients (48.79%)with no depression,mild depression in 78 cases (20.91%),moderate depression in 59 cases (15.82%),severe depression in 54 cases (14.48%). 51.21% patients had different degree of depression.Of the 373 patients,there were 123 patients (32.97%)with no anxiety,mild anxiety in 141 cases (37.80%),mild and moderate anxiety in 55 cases (14.75%),moderate anxiety in 33 cases (8.85%),severe anxiety in 21 cases (5.63%).A total of 67.03% patients had different degrees of anx-iety.Huzhou city maintenance hemodialysis,female patients had depression rate of 80.70%,anxiety rate of 65.50%, which were significantly higher than the rate of depression 55.45%,anxiety rate 39.11% in male patients,the differ-ences were statistically significant (P <0.01 -0.05).In maintenance hemodialysis patients,the anxiety and depres-sion status gradually increased with age.Severe malnutrition in maintenance hemodialysis patients,the rate of depres-sion,anxiety rates were 80.65%,90.32%,the rates of depression,anxiety were 44.49%,61.81% in normal and mild malnutrition patients,the differences were statistically significant (all P <0.01).SF -36 scale showed that most of the patients with maintenance hemodialysis patients had poor quality of life,and the function was limited. Conclusion Maintenance hemodialysis patients,the general existence of anxiety and depression in the mental state in Huzhou city.Moreover,the psychological state of the patients is influenced by sex,age,nutrition status,and other factors.In clinical,it is needed to provide effective psychological intervention for patients to improve the quality of life,according to the psychological status and related factors of maintenance hemodialysis patients.