Hirudin (HV) is known as the most potent and specific inhibitor of thrombin. Although hirudin has many advantages , it has the bleeding side effect and this is the great shortage of hiudin for clinical application. In order to alleviate bleeding side effect of hirudin, fusion protein, named as FHV (fusion hirudin linked with FXa recognition peptide) was designed. The fusion protein gene ( fhv) was cloned into plasmid pPIC9K. FHV engineered Pichia pastoris containing high copies was chosen for fermentation and purification at 30 L fermentor scale, finally, FHV with purity of above 97% was obtained. To investigate the function of FHV in vivo, mouse tail thrombosis model was used. In the mice thrombus tail model induced by carrageenan, FHV decreased the length of tail thrombus significantly, similar to that of HV control, and had no obvious effects on the TT, PT and APTT. In conclusion, FHV is constructed and expressed in yeast. FHV fusion proteins is obtained by fermentation and purification. FHV has antithrombotic effects not influencing IT, PT and APTT after administration immediately in animal models. Therefore, FHV is a promising anticoagulant and antithrombotic drug.

Objective To explore the possible mechanism and protective effect of Xuebijing injection (血必净注射液)and dexamethasone on rats with paraquat-induced chronic pulmonary injury.Methods Thirty male Wistar rats were randomly divided into six groups:normal group(n=5),model group(n=5), treatment groups(n=20).In the normal group,normal saline was used,while in the other groups,20% paraquat 80 mg/kg was injected peritoneally for poisoning.After 2 hours of intoxication,low dose Xuebijing injection(1.25 g/kg),high dose Xuebijing injection(2.50 g/kg),dexamethasone(25 mg/kg),high dose Xuebijing injection combined with dexamethasone(combined group)respectively were administered into the four different treatment groups,equal amount of normal saline was given to the normal and model groups,and the treatment continued for 4 days.At 28 days after paraquat injection,5 rats in each group were killed respectively,serum transforming growth factor-?1(TGF-?1)and hydroxyproline(HYP)level in the lung homogenate were measured,and pulmonary coefficient and histological changes were observed.Results In the treatment groups,the levels of serum TGF-?1 and lung tissue HYP,pulmonary coefficient were leas than those of model group,and among the treatment groups,combined group had the best results(all P

OBJECTIVETo analyze the factors affecting pathologic complete response (pCR) to neoadjuvant chemotherapy (NAC) in breast cancer patients.

METHODSA retrospective cohort study was carried out to analyze the clinical data of 141 breast cancer patients treated with neoadjuvant chemotherapy. The factors affecting pCR and the changes of tumor receptor status before and after treatment were analyzed.

RESULTSAmong all the 141 patients, 21 patients (14.9%) achieved pCR. The rate of pCR achieved by regimens of anthracycline combined with taxane was higher (16.8%, 19/113) than that by anthracycline-containing regimens (7.1%, 1/14). The dose intensity of anthracycline had a significant correlation with pCR rate (P < 0.05). The pCR rate in the relative dose intensity of taxane ≥ 0.85 arm was higher than that of < 0.85 arm (P = 0.02). Eighty patients (56.7%) had completed more than 4 cycles of chemotherapy and the median time to achieve pCR was 6 (3 to 10) cycles. The pCR rate had a significant difference between patients < 6 and ≥ 6 cycles (7.1% vs. 22.5%,P = 0.01). Multivariate analysis showed that tumor size measured by palpation ≤ 5 cm and ≥ 6 chemotherapy cycles were significantly related with pCR rate (P < 0.05). In all the 21 pCR patients, the pre-treatment ER(-), PR(-), HER-2(-) statuses were in 14, 14 and 17 patients, respectively. The status of ER, PR, HER-2 of most patients (74.2%, 69.7% and 87.7%, respectively) was not changed after treatment. Among the patients with changes in receptor status, ER changed from negative to positive was in the majority (37.1%, 13/35 vs. 12.9%, 4/31, P < 0.05), and the percentage of changes in PR and HER-2 status had no significant differences.

CONCLUSIONSThe regimens of anthracycline combined with taxane can achieve a higher pCR rate. The lymph node and receptor status before therapy have no significant correlation with pCR. Patients who have primary tumor size ≤ 5 cm, ≥ 6 chemotherapy cycles and enough dose intensity are easier to achieve pCR. The receptor status before and after therapy should be determined, and according to any positive results, physicians can chose HER-2 targeted therapy and/or endocrine therapy after surgery to benefit the patients.

Adult ; Aged ; Aged, 80 and over ; Anthracyclines ; administration & dosage ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; therapeutic use ; Breast Neoplasms ; drug therapy ; metabolism ; pathology ; Bridged-Ring Compounds ; administration & dosage ; Chemotherapy, Adjuvant ; Dose-Response Relationship, Drug ; Female ; Humans ; Lymphatic Metastasis ; Middle Aged ; Neoadjuvant Therapy ; methods ; Proportional Hazards Models ; Receptor, ErbB-2 ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Remission Induction ; Retrospective Studies ; Taxoids ; administration & dosage ; Tumor Burden

BACKGROUND AND OBJECTIVECybeKnife is a newly developed technology in the field of stereotactic radiosurgery/radiotherapy (SRS/SRT). Compared with conventional SRS/SRT, there are many advantages for CyberKnife in terms of treating tumors that move with respiration, being real-time image-guidance, frameless, high accurateness, and so on. Recently, it has been used to treat different types of malignant carcinoma including intracranial and caudomedial tumors. This study was designed to evaluate the short-term efficacy and toxicity of the CyberKnife radiotherapy for locally advanced pancreatic cancer.

METHODSA total of 20 patients with locally advanced (stage II-III) pancreatic cancer treated with CyberKnife were recruited between April 2009 and December 2009. Of 20 patients, 13 were with cancer located at the pancreatic head and 7 were located at the pancreatic body and tail. The planning target volume (PTV) was defined as gross tumor volume (GTV) plus 2-3 mm, and more than 95% PTV should be covered by 75% isodose surface. The median of PTV was 47 cm³ (26-64 cm³). The median total prescription dose was 40 Gy (32-55 Gy) at 3-6 fractions. During treatment delivery, X-Sight Spine Tracking System was used in 5 patients to track movement of the tumor. Other 15 patients were implanted fiducials in the tumors to track movement of the tumor and patient breathing patterns.

RESULTSThe median follow-up time was 7 months (3-11 months). All patients had finished the treatment and 19 were alive by the last follow-up. Slight fatigue was the most common complain. Evaluated by CT scan, 6 were complete response, 9 were partial response, 3 were stable disease, and 1 was progression; 1 was dead. There were 6 patients with grade I granulocytopenia, 7 with grade I nausea, and 5 with grade II vomiting.

CONCLUSIONSThe CyberKnife radiosurgery for the locally advanced pancreatic cancer shows a high rate of local control and minimal toxicity. Long-term follow-up is necessary to evaluate the survival and late toxicity.

Adult ; Aged ; CA-19-9 Antigen ; blood ; Female ; Follow-Up Studies ; Humans ; Leukopenia ; etiology ; Male ; Middle Aged ; Nausea ; etiology ; Neoplasm Staging ; Pancreatic Neoplasms ; diagnostic imaging ; surgery ; Quality of Life ; Radionuclide Imaging ; Radiosurgery ; adverse effects ; Radiotherapy Dosage ; Remission Induction ; Thrombocytopenia ; etiology

To combine the fibrinolytic with anticoagulant activities for therapy of thrombotic deseases, a fusion protein made of tissue-type plasminogen activator (t-PA) and hirudin was constructed and expressed in chia pastoris. To improve thrombolytic properties of t-PA and reduce bleeding side effect of hirudin, FXa-recognition sequence was introduced between t-PA and hirudin molecules.The anticoagulant activity of hirudin can be target-released through cleavage of FXa at thrombus site. t-PA gene and hirudin gene with FXa-recognition sequence at its 5'-terminal were obtained by RT-PCR and PCR respectively. The fusion protein gene was cloned into plasmid pIC9K and electroporated into the genome of Pichia pastoris GS115. The expression of fusion protein was induced by methanol in shaking flask and secreted into the culture medium. Two forms of the fusion protein, single-chain and double-chain linked by a disulfide bond (due to the cleveage of t-PA at Arg275-Ile276), were obtained. The intact fusion protein retained the fibrinolytic activity but lacked any anticoagulant activity. After cleavage by FXa, the fusion protein liberated intact free hirudin to exert its anticoagulant activity. So, the fusion protein is a bifunctional molecule having good prospect to develop into a new targeted therapeutic agent with reduced bleeding side effect for thrombotic diseases.
Animals ; Cloning, Molecular ; Electroporation ; Hirudins ; biosynthesis ; genetics ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Plasminogen Activator ; biosynthesis ; genetics

Objective To study the correlation between cognitive dysfunction and cholinergic neuron changes in basal forebrain(BFB)and brainstem reticular formation(BSRF)areas after brain con- cussion(BC)in rats.Methods Eighty-four Spragne-Dawley rats weighing 250-310g were used.The BC rat models were reproduced by a self-made simple pendulum impact device,then the rats were ran- domized into one control group and six experimental groups(1 day,2 day,4 day,8 day,16 day,and 24 day groups;n=12 in each group).A Morris Water Maze(MWM)test was used to assess learning and memory function of the rats.Cholinergic neurons in the BFB and BSRF were identified with choline acetyltransferase(CHAT)antibody and quantitated.Results Compared with the control group,the la- tency to find the platform in MWM was much longer on days 1-3 after BC,but there was no statistical difference on days 4-21 after BC.The cell number and the ChAT expression activity of cholinergic neu- rons in the BFB were obviously decreased after BC,and reached the lowest level at 8 days after BC,then increased gradually and nearly reached the normal level at 24 days.The ChAT expression activity in BSRF declined on the first 2 days after BC,then went up gradually,and reached the peak at the 24th day.Conclusion The spatial cognition deficit of BC rats can be detected by MWM in the early period (1-3 days after BC).There are significant changes in the number and ChAT expression activity in BFB and BSRF after BC.The change of cholinergic neurons may be correlated with cognitive deficits in BC rats.

OBJECTIVETo compare the efficiency of response evaluation by clinical examination, ultrasonograghy and mammography in neoadjuvant chemotherapy (NAC) for breast cancer.

METHODSA retrospective cohort study was conducted to analyze the data of 141 patients treated with neoadjuvant chemotherapy. Response evaluation was performed by clinical palpation, ultrasound and mammography.

RESULTSOnly 12 (8.5%) among the 141 patients presented with a stage I tumor. The tumor size determined by palpation was often larger than that by ultrasound before therapy (P < 0.01). Among patients with suspicions axillary nodes checked by ultrasound, 88.3% (53/60) of them had positive nodes by pathology before NAC, and 34.5% (10/29) of patients with negative nodes determined by ultrasound had positive nodes by pathology. In all the 141 patients, 21(14.9%) showed pathological complete remission in both the primary tumor and lymph node. For response evaluation, the false complete remission rate judged by clinical examination was 46.8% (22/47), and the false tumor residual rate by ultrasound was 84.0% (21/25). In 53.5% (23/43) of patients the response could not be assessed by mammography due to that the tumors were undistinguishable in size. The range of microcalcification was not reduced in 5 patients with a partial response of the tumor. 25 patients experienced needle puncture during therapy. Among them, in the 9 pathologically negative patients, only 3 achieved pCR, and the other 16 positive patients didn't achieve pCR.

CONCLUSIONUsing the puncture or sentinel lymph node biopsy, clinicians should pay enough emphasis on the pathological determination of the node status before chemotherapy. Clinicians will make a quite of false judgment of the tumor by clinical examination, ultrasound or mammography. They may use needle puncture during therapy to evaluate the response of neoadjuvant chemotherapy, and the result should be analyzed synthetically.

Adult ; Aged ; Aged, 80 and over ; Axilla ; Breast Neoplasms ; diagnostic imaging ; drug therapy ; pathology ; Carcinoma, Ductal, Breast ; diagnostic imaging ; drug therapy ; pathology ; Chemotherapy, Adjuvant ; Cohort Studies ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Mammography ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Staging ; Remission Induction ; methods ; Retrospective Studies ; Sentinel Lymph Node Biopsy ; Ultrasonography

Objective To compare the effects of valsartan combined with amlodipine or hydrochlorothiazide regimen on blood pressure variation and plasma nitric oxide(NO)and endothelin(ET)in elderly hypertensive patients.Methods A total of 61 elderly patients with grade 2 or 3 hypertension were randomized into valsartan + amlodipine(the amlodipine group,n =31)or valsartan + hydrochlorothiazide (the hvdrochlorothiazide group,n =30)group.Blood lipids,fasting plasma glucose and uric acid were determined before the treatment.24-hour dynamic blood pressure,NO and ET were monitored at baseline,8 and 16 weeks after treatment.Results 24 hours blood pressure and daytime blood pressure were similarbetween two groups at all 3 time points.At 16 weeks,morning systolic blood pressure surge was significantly lower in amlodipine group than in hydrochlorothiazide group[(22.6 ±8.8)mm Hg(1 mm Hg =0.133 kPa)vs.(26.3 ± 13.7)mm Hg,P ＜ 0.05].24 hours systolic blood pressure variability(SBPV)decreased progressively in both groups[the amlodipine group:(12.5 ±2.8)mm Hg vs.(10.2 ±2.2)mm Hg vs.(8.8 ±1.6)mm Hg,P ＜0.01;the hydrochlorothiazide group:(12.5 ±2.5)mm Hg vs.(10.7 ±2.2)mm Hg vs.(9.6 ±2.0)mm Hg,P ＜ 0.01].Daytime SBPV also decreased progressively in both groups[the amlodipinegroup:(12.2±3.0)mm Hgvs.(10.1 ±2.3)mm Hgvs.(8.4±1.9)mm Hg,P＜0.01;the hydrochlorothiazide group:(11.8 ± 2.7)mm Hg vs.(10.4 ± 1.9)mm Hg vs.(9.6 ± 2.2)mm Hg,P ＜0.01].24 hours diastolic blood pressure variability(DBPV)was significantly reduced post therapy in the amlodipine group[(15.5 ±3.4)mm Hgvs.(13.0±3.5)mm Hgvs.(12.3±2.5),P＜0.01]but not in the hydrochlorothiazide group.NO increased progressively[(27.3 ± 13.6)μmol/L vs.(47.2 ± 16.3)μmol/L vs.(69.5 ± 18.9)μmol/L in the anlodipine group,P ＜ 0.0l ;(33.5 ± 13.9)μmol/L vs.(49.7 ±21.9)μmol/L vs.(66.7 ± 24.7)μmol/L in the hydrochlorothiazide group,P ＜ 0.01]and ET decreased progressively[(45.3 ±8.0)ng/L vs.(37.4 ±3.9)ng/L vs.(34.2 ±4.4)ng/l in the arnlodipine group,P ＜0.01 ;(46.6 ± 10.4)ng/L vs.(37.0 ± 5.4)ng/L vs.(36.1 ± 8.2)ng/L in the hydrochlorothiazide group,P ＜ 0.01]in both groups.Conclusion Valsartan in combination with amlodipine or hydroehlorothiazide can both effectively lower BPV in elderly hypertensive patients and improve the vascular endothelial function and the former regimen is more suitable for elderly hypertensive patients.

OBJECTIVETo evaluate the correlation of clinical effects and reasonable doses of docetaxel salvage therapy for patients with metastatic breast cancer.

METHODSWe reviewed retrospectively the clinical records of patients with metastatic breast cancer treated with docetaxel and statistically analyzed the correlation between clinical effects and reasonable doses of docetaxel.

RESULTSThe objective response rate and clinical benefit rate of docetaxol in patients with metastatic breast cancer were 27.0% and 35.0%, respectively, and the median progression free survival (PFS) was 5.0 (3.8 - 6.3) months. In the analysis at a single dose level, the clinical benefit rate and PFS of the ≥ 90.0 mg/m(2) docetaxel group were superior to that of the < 90.0 mg/m(2) group (P = 0.008, P = 0.045). Multi-dose level group stratified analysis showed that the docetaxel < 75.0 mg/m(2) group was better than the 75.0 - 84.9 mg/m(2) PFS group (P = 0.018), and the ≥ 95.0 mg/m(2) group was better than the 75.0 - 84.9 mg/m(2) group (P = 0.048). In patients who received >third line treatment or previously received paclitaxel adjuvant therapy, the PFS of the ≥ 94.9 mg/m(2) docetaxel group was 6.0 months, better than the 3.0 months of the 75.0 ∼ 84.9 mg/m(2) group (P = 0.031; P = 0.021).

CONCLUSIONThere is a clear correlation between clinical effects and reasonable doses of docetaxel salvage therapy in patients with metastatic breast cancer.

Adult ; Aged ; Antineoplastic Agents ; administration & dosage ; therapeutic use ; Bone Neoplasms ; drug therapy ; secondary ; Breast Neoplasms ; drug therapy ; pathology ; Disease-Free Survival ; Dose-Response Relationship, Drug ; Drug Administration Schedule ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; drug therapy ; secondary ; Lung Neoplasms ; drug therapy ; secondary ; Middle Aged ; Remission Induction ; Retrospective Studies ; Salvage Therapy ; Taxoids ; administration & dosage ; therapeutic use ; Young Adult

This study was aimed to investigate the cytogenetic changes of MDS cell line (MUTZ-1) with chromosome 5q deletion. R-banding analysis was used to identify chromosome aberrations in MDS cell line and Vysis Spectra Vysion M-FISH was used to further characterize chromosomal complex karyotype. The results indicated that M-FISH exhibited obvious chromosomal aberrations with high frequency including translocation, insertion, breakage and rearrangement, deletion and increasement of chromosome number, the complex karyotype of MUTZ-1 was shown as 50, xx, der (1) t (1;2), ins (1;14), +der (2) t(2;19), der (3) t (3;5), der (3) (3::5::22), 5q-, der (6) t (3;6), der (7) (18::7::17), +8, +der (9) t (1;9), der (10) t (1;10), +11, +12, der (?13) (10::13::5::8), der (14) t (8;14), der (14) t (14, 15), der (15) t (15;21) x 2, +17, +18, -21, -22. It is concluded that M-FISH analysis revealed obvious changes in complex karyotype of MDS cell line MUTZ-1, and the M-FISH technique can increase accuracy of detection for chromosomal complex karyotype, and help diagnosis and prognostic evaluation of MDS.
Child, Preschool ; Chromosome Aberrations ; Chromosome Deletion ; Chromosomes, Human, Pair 5 ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Karyotyping ; Myelodysplastic Syndromes ; genetics ; Translocation, Genetic ; Tumor Cells, Cultured