Alfimeprase(ALF)is a recombinantly modified variant of non-hemorrhagic zinc metalloproteinase fibrolase.The target gene alf was obtained from the clone vector p43-alf and cloned into the Pichia pastoris expression vector pPICZ? A.Through high efficiency transformation and Zeocin selection,the recombinant strains of pPICZ?A-alf /GS115 were isolated.In order to achieve a high level expression of recombinant Alfimeprase(rALF),optimization of pH value,methanol daily addition concentration,cell density and methanol induction time points were carried out,and the production of rALF reached up to 425 mg/L.By His?Bind chromatography,the purity of secreted rALF was as high as 95 %.SDS-PAGE and Western blot analysis show that rALF has a molecular weight of about 24 kDa and is bound specifically to anti-His?tag monoclonal antibody.Activity identification results of the modified fibrin plate method demonstrate that the secreted rALF has high fibrinolytic activity.Thus sets up an optimized expression system for ALF,which will play an important role in its further studies and industrial production.

BACKGROUND AND OBJECTIVEAnemia can not only reduce the quality of life of patients with cancer, but also affect their survival. This study was to investigate the prognostic value of hemoglobin (Hb) level in patients with nasopharyngeal carcinoma (NPC) treated with radiotherapy.

METHODSClinical data of 520 NPC patients received definitive radiotherapy between 2000 and 2002 at Sun Yat sen University Cancer Center were analyzed. Patients were stratified into normal Hb level and anemia groups according to their Hb levels before, during, and after radiation. Anemia was defined according to World Health Organization criteria as Hb level < 130 g/L in men and < 120 g/L in women. Hb continuous decrease group and non decrease group were defined according to Hb changes in the patients during radiotherapy. Loco regional recurrence free survival (LRFS) and overall survival (OS) rates were estimated using the Kaplan Meier method. Multivariate analysis was performed using the Cox model to analyze the prognostic factors.

RESULTSBefore radiation, the 5 year LRFS rates were 60.9% in anemia group and 63.9% in normal Hb level group (P = 0.337); the 5 year OS rates were 65.2% and 71.0%, respectively (P = 0.299). During radiation, the 5 year LRFS rates were 56.7% in anemia group and 67.9% in normal Hb level group (P = 0.013); the 5 year OS rates were 61.0% and 75.9%, respectively (P = 0.001). After radiation, the 5 year LRFS rates were 59.6% in anemia group and 64.9% in normal Hb level group (P = 0.169); the 5 year OS rates were 65.0% and 71.9%, respectively (P = 0.090). The 5 year LRFS and OS rates were significantly lower in Hb continuous decrease group than in Hb non decrease group (59.1% vs. 69.3%, P = 0.032; 66.2% vs. 76.4%, P=0.011). Multivariate analysis showed that the continuous decrease of Hb was an independent prognostic factor for OS.

CONCLUSIONThe change in Hb level during radiotherapy is an important prognostic factor affecting the OS of NPC patients.

Adolescent ; Adult ; Age Factors ; Aged ; Anemia ; blood ; etiology ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma ; Cisplatin ; administration & dosage ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Hemoglobins ; metabolism ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; blood ; drug therapy ; radiotherapy ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Particle Accelerators ; Proportional Hazards Models ; Radiotherapy, High-Energy ; adverse effects ; Survival Rate ; Young Adult

OBJECTIVEMultivariate analysis was used to select the risk factors in non-insulin dependent diabetes mellitus (NIDDM) patients with oral candidosis, and to establish the forecasting equation, aimed to detect the risk of oral candidosis among NIDDM patients.

METHODS140 NIDDM patients were included in this study. 11 clinical parameters including gender, age, course smoking, fasting blood glucose, oral hygiene status, systemic manifestation, oral mucous membrane status, and denture were recorded respectively. Oral rinse technique was used to detect the salivary candidal carriage. The isolates were identified using CHROM agar Candida test. The Logistic multivariate regression analysis was carried our for risk factors analysis.

RESULTSCandida was found in 69 out of 140 NIDDM cases, and Candida albicans was the major species isolated. The poor glycemic control, poor oral hygiene, and dry mouth were the risk factors of oral candidosis in NIDDM patients, and the forecasting equation was established. Using substitution method, the veracity of the forecasting equation was 82.1%.

CONCLUSIONPoor glycemic control, poor oral hygiene and dry mouth were risk factors of oral candidosis among NIDDM patients. The probability obtained from the forecasting equation may offer references for predicting and preventing the oral candidosis in NIDDM patients.

Adult ; Candida ; Candida albicans ; Candidiasis, Oral ; Diabetes Mellitus, Type 1 ; Diabetes Mellitus, Type 2 ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Oral Hygiene ; Risk Factors ; Smoking

BACKGROUNDThe relationship between monosymptomatic resting tremor (mRT) and Parkinson's disease (PD) remains controversial. In this study, we aimed to assess the function of presynaptic dopaminergic neurons in patients with mRT by dopamine transporter positron emission tomography (DAT-PET) and to evaluate the utility of clinical features or electrophysiological studies in differential diagnosis.

METHODSThirty-three consecutive patients with mRT were enrolled prospectively. The Unified Parkinson's Disease Rating Scale and electromyography were tested before DAT-PET. Striatal asymmetry index (SAI) was calculated, and a normal DAT-PET was defined as a SAI of <15%. Scans without evidence of dopaminergic deficits (SWEDDs) were diagnosed in patients with a subsequent normal DAT-PET and structural magnetic resonance imaging.

RESULTSTwenty-eight mRT patients with a significant reduction in uptake of DAT binding in the striatum were diagnosed with PD, while the remained 5 with a normal DAT-PET scan were SWEDDs. As for UPRDS, the dressing and hygiene score, walking in motor experiences of daily living (Part II) and motor examination (Part III) were significant different between two groups (P < 0.05 and P < 0.01, respectively). Bilateral tremor was more frequent in the SWEDDs group (P < 0.05). The frequency of resting tremor and the amplitude of postural tremor tend to be higher in the SWEDDs group (P = 0.08 and P = 0.05, respectively).

CONCLUSIONSmRT is heterogeneous in presynaptic nigrostriatal dopaminergic degeneration, which can be determined by DAT-PET brain imaging. Clinical and electrophysiological features may provide clues to distinguish PD from SWEDDs.

Adult ; Aged ; Dopamine Plasma Membrane Transport Proteins ; Female ; Humans ; Male ; Middle Aged ; Parkinson Disease ; diagnosis ; Positron-Emission Tomography ; methods ; Prospective Studies ; Tremor ; diagnosis

OBJECTIVETo provide the foundation for reasonable utilization by analysing the essential oils of Flos chrysanthemi Indici in different areas.

METHODThe essential oils were extracted by using steam distillation and separated with GC capillary columns. The components were quantitatively determined with normalization method, and were identified with GC-MS.

RESULTS18, 17 and 20 compounds of essential oils from Guangxi, Guangdong and Hubei were identified.

CONCLUSIONThere are significant differences among the components and contents of essential oils of Flos chrysanthemi Indici from Guangxi, Guangdong and Hubei.

Chrysanthemum ; chemistry ; Climate ; Ecology ; Flowers ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Seasons

Objective To explore inhibition of amlodipine on myocar-dial cell injury induced by ischemia reperfusion .Methods Thirty Wistar rats were randomly into three groups: sham operation group ( n =10 ) , model group group ( n =10 ) , test group ( 2 mg? kg -1 amlodipine , n=10).The model group and test group were made by ligation of left anterior descending coronary artery to make the model of myocardial ischemia reperfusion.Rats in each group were administered 7 d before ligation.Cell apoptosis was examed by flow dual staining method .The activity of cysteinyl aspartate specific proteinase 3 ( Caspase 3 ) was measured by spectrophotometry .The activation of phosphatidylinositol 3 kinase ( PI3K)/protein kinase B ( AKT) signalling pathway, B-cell lymphoma-2 ( Bcl-2 ) , Bcl-2 associated X protein ( Bax ) were assayed by Western blot .Results Compared to sham operation group on early and late myocardial cell apoptosis , myocardial cell apoptosis with ( 2.34 ±0.35 )%, (3.58 ±0.39 )%, that on early and late myocardial cell apoptosis were increased with ( 15.69 ±1.14 )%, (24.74 ±2.56)%in model group ( P <0.05 ) .Compared to sham operation group on the activity of Bax with (0.18 ±0.01) and Caspase 3 activity with (1.00 ±0.10), the expression of Bax express with (0.62 ±0.06) and Caspase 3 activity with (3.98 ±0.18) in model group were increased (P<0.05).Compared to sham operation group on the expre-ssion of Bcl-2 with (0.99 ±0.10 ) and expression of PI3K with (0.89 ±0.06 ), on the expression of Bcl-2 with (0.14 ±0.01) and expression of PI3K with (0.18 ±0.01) in model group were decreased (P<0.05). Compared to sham operation group on phosphorylation of AKT with ( 0.95 ±0.10 ) , the phosphorylation of AKT in model group with (0.13 ±0.01 ) was decreased ( P<0.05 ).Compared with model group , test group could change the variation on the early and late myocardial cell apoptosis with ( 5.23 ±0.13 )%, ( 8.09 ±0.35 )% while on the Caspase 3 activity with ( 1.47 ±0.14 ) ( P<0.05 ) .Conclusion These results suggested that amlodipine inhibited myocardial ischemia reperfusion injury, which was related to activition PI3K/AKT signal pathway.

The advantages of Canon Aquilion One CT scanner over the traditional CT machines were summarized.Three cases of scanning table faults of Canon Aquilion One CT scanner were introduced in terms of cause and specific troubleshooting method.References were provided for medical engineers to treat similar faults.[Chinese Medical Equipment Journal,2023,44(9):114-116]

The advantages of Toshiba Aquilion series spiral CT were described over the traditional CT.One case of MR sensor fault of Toshiba Aquilion 16 CT and its cause was introduced,and the countermeasure was proposed accordingly.References were provided for medical engineers to treat similar faults.[Chinese Medical Equipment Journal,2024,45(2):118-120]

Purpose: Gastric cancer (GC) is the second most lethal cancer globally and is associated with poor prognosis. Fatty acid-binding proteins (FABPs) can regulate biological properties of carcinoma cells. FABP5 is overexpressed in many types of cancers; however, the role and mechanisms of action of FABP5 in GC remain unclear. In this study, we aimed to evaluate the clinical and biological functions of FABP5 in GC. Materials and Methods: We assessed FABP5 expression using immunohistochemical analysis in 79 patients with GC and evaluated its biological functions following in vitro and in vivo ectopic expression. FABP5 targets relevant to GC progression were determined using RNA sequencing (RNA-seq). Results: Elevated FABP5 expression was closely associated with poor outcomes, and ectopic expression of FABP5 promoted proliferation, invasion, migration, and carcinogenicity of GC cells, thus suggesting its potential tumor-promoting role in GC. Additionally, RNA-seq analysis indicated that FABP5 activates immune-related pathways, including cytokinecytokine receptor interaction pathways, interleukin-17 signaling, and tumor necrosis factor signaling, suggesting an important rationale for the possible development of therapies that combine FABP5-targeted drugs with immunotherapeutics. Conclusions These findings highlight the biological mechanisms and clinical implications of FABP5 in GC and suggest its potential as an adverse prognostic factor and/or therapeutic target.

BACKGROUNDTransplantation of adult bone marrow-derived mesenchymal stem cells (MSCs) has been proposed as a strategy for cardiac repair following myocardial damage. However cell transplantation strategies to replace lost myocardium are limited by the inability to deliver large numbers of cells that resist peritransplantation graft cell death. Accordingly, we set out to isolate and expand adult swine bone marrow-derived MSCs, and to engineer these cells to overexpress AKT1 (protein kinase B), to test the hypothesis that AKT1-engineered MSCs are more resistant to apoptosis and can enhance cardiac repair after transplantation into the ischemic swine heart.

METHODSThe CDS (regulation domain of AKT1) AKT1-cDNA fragment was amplified, and MSCs were transfected following synthesis with a pCDH1-AKT1 shuttling plasmid. Western blotting analysis and real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed. Myocardial infarction (MI) models were constructed in Meishan pigs, and cardiac function was evaluated by magnetic resonance imaging (MRI) measurements and echocardiography 4 weeks later. All pigs were assigned to four groups: control (A), DMEM (B), MSC (C), and AKT-transfected (D). MSCs were transfected with the AKT1 gene, and autologous BrdU-labeled stem cells (1 x 10(7)/5 ml) were injected into left anterior descending coronary atery (LAD) of the infarct heart in groups C and D. In group B, DMEM was injected using the same approach. In group A, there was no injection following LAD occlusion. After 4 weeks, cardiac function and regional perfusion measurements were repeated by MRI and echocardiography, and histological characteristics of the hearts were assessed. Connecxin-43 (CX-43), BrdU, and von Willebrand factor (VWF) immunoreactivity was tested using enzyme linked immunosorbent assay (ELISA). Vascular endothelial growth factor (VEGF), transforming growth factor-beta1 (TGF-beta1) were analyzed at the same time.

RESULTSAKT1-cDNA was cloned into pCDH1-MCS1-EF1-copGFP and the sequence was confirmed. AKT mRNA expression was detected at 24 hours after transfection. AKT1 expression in MSCs remained strong after 2 weeks, according to real-time RT-PCR and Western blotting. Prior to cell implantation, end-diastolic left ventricular dimension (EDLVd) increased and stroke volume (SV) decreased in the MI hearts. MRI scans revealed significantly improved cardiac function following implantation, and implanted MSCs prevented thinning and expanding in the infarct region, as well as improved contraction and increased perfusion in all groups compared to control hearts. The left ventricular chamber size was smaller in cell-transplanted hearts than in control hearts. Moreover, group D exhibited significant improvement. The expression of CX-43, BrdU, and VWF could be found in the immunohistochemical pathological sections of group C and group D. The level of VEGF reached a high level 1 week after implanting the MSCs, but the level of TGF-beta1 decreased gradually.

CONCLUSIONSThe AKT1-expressing lentiviral vector resulted in stable over-expression of AKT1 in MSCs. MSC engraftment in host myocardium improved cardiac function by attenuating contractile dysfunction and pathological thinning of the infracted left ventricular wall, which likely resulted from myocardial regeneration and angiogenesis.

Animals ; Blotting, Western ; Disease Models, Animal ; Electrophoresis ; Genetic Vectors ; genetics ; Lentivirus ; genetics ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Myocardial Infarction ; metabolism ; therapy ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Swine ; Ventricular Remodeling ; physiology