OBJECTIVETo construct plant expression pCAMBIA1301-PMI by substituting PMI for hygromycin resistance gene in pCAMBIA1301 and obtain transgenic Salvia miltiorrhiza f. alba using PMI-mannose selection system.

METHODThe 6-phosphomannose isomerase gene (PMI) of Escherichia coli was amplified by PCR. Sequence analysis showed that it shared 100% amino acids identities with the sequences of PMI genes isolates reported in the NCBI. Based on pCAMBIA1305, the plant expression pCAMBIA1305-PMI was constructed successfully by substituting PMI for hygromycin resistance gene in pCAMBIA1305. pCAMBIA1305-PMI was transformed into Agrobacterium tumefaciens LBA4404, and then the leaves of S. miltiorrhiza f. alba were inoculated in LBA4404 with pCAMBIA1305-PMI.

RESULTPlant expression pCAMBIA1301-PMI was successfully constructed and the leaves of S. miltiorrhiza f. alba inoculated in LBA4404 with pCAMBIA1305-PMI were selected on medium supplemented with a combination of 20 g x L(-1) mannose and 10 g x L(-1) sucrose as a carbon source. The transformation efficiency rate was 23.7%.

CONCLUSIONGenetic transformation was confirmed by PCR, indicating that a new method for obtaining transgenic S. miltiorrhiza f. alba plants was developed using PMI-mannose selection system.

Anti-Bacterial Agents ; pharmacology ; Biomarkers ; Cinnamates ; pharmacology ; Escherichia coli ; enzymology ; genetics ; Escherichia coli Proteins ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Hygromycin B ; analogs & derivatives ; pharmacology ; Mannose-6-Phosphate Isomerase ; genetics ; metabolism ; Plants, Genetically Modified ; drug effects ; genetics ; metabolism ; Salvia miltiorrhiza ; drug effects ; genetics ; metabolism ; Transformation, Genetic

OBJECTIVETo study the application of degrading multi-enzymes from Ganoderma lucidum in extracting effective constituents from fibrous roots of Salvia miltiorrhiza.

METHODEffective constituents were extracted from fibrous roots by degrading multi-enzymes of wood fiber. The enzymatic parameters were optimized by the orthogonal design.

RESULTThe extraction efficiencies of total tanshinones and total salvianolic acids in the extracts of fibrous roots of S. miltiorrhiza was obtained using optimum enzymolysis process reached 11.923%, 12.465%, respectively, which were 62.794%, 56.086% more than that by conventional non-enzymatic hydrolysis.

CONCLUSIONDegrading multi-enzymes of wood fiber can be used to fully extract effective constituents from fibrous roots of S. miltiorrhiza, which provides a new approach for recycling wastes of traditional Chinese medicines.

Alkenes ; isolation & purification ; metabolism ; Diterpenes, Abietane ; isolation & purification ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; metabolism ; Hydrogen-Ion Concentration ; Plant Roots ; chemistry ; Polyphenols ; isolation & purification ; metabolism ; Reishi ; enzymology ; Salvia miltiorrhiza ; chemistry ; Temperature ; Wood ; enzymology

OBJECTIVE: To establish a diagnostic model for diffuse axonal injury (DAI) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). To screen the proteins or peptides associated with DAI for providing the biomarkers with theoretic foundation. METHODS: Fifteen male Sprague-Dawley rats were randomly divided into DAI group (n = 10) and control group (n = 5). The protein or peptide expression profiles of rat brain stem were detected by MALDI-TOF-MS. ClinProTools 2.2 software was used to find specific peaks, and a diagnostic model was established by the genetic algorithm. RESULTS: There were significant differences in 61 peaks of DAI group (P < 0.05), 9 peaks were down-regulated and 52 up-regulated. The diagnostic model was established based on 5 different peaks. The specificity and sensitivity of cross validation was 96.14% and 95.98%; while the specificity and sensitivity of blind validation showed was 73.33% and 70.00%, respectively. CONCLUSION A specific and sensitive diagnostic model of DAI can be established by MALDI-TOF-MS to provide a potential value for determining DAI in forensic practice.
Animals ; Biomarkers ; Brain Stem/metabolism* ; Diffuse Axonal Injury/diagnosis* ; Down-Regulation ; Male ; Peptides/blood* ; Proteomics ; Rats ; Rats, Sprague-Dawley ; Sensitivity and Specificity ; Software ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods* ; Up-Regulation

Matrix-assisted laser desorption/ionization time-of-flight imaging mass spectrometry (MALDI-TOF-IMS) has been a classical technique for studying proteomics in present and a tool for analyzing the distribution of proteins and small molecules within biological tissue sections. MALDI-TOF-IMS can analyze multiple unknown compounds in biological tissue sections simultaneously through a single measurement which can obtain molecule imaging of the tissue while maintaining the integrity of cellular and molecules in tissue. In recent years, imaging mass spectrometry technique develops relatively quickly in all biomedical domain. This paper based on the relevant data and reviews the present developing level of MALDI-TOF-IMS, the principle of imaging mass spectrometry, methology and the prospect in forensic pathology.
Diagnostic Imaging ; Forensic Sciences/methods* ; Humans ; Male ; Proteins ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo explore the causes of the formation of traumatic carotid-cavernous fistulas and the therapeutic effect of detachable balloon and/or coil embolization and the prevention of its complications.

METHODSFrom October, 1992 to March, 2002, 17 patients with traumatic carotid-cavernous fistulas were treated with detachable balloon and/or coil embolization in our hospital. The clinical data and imaging features of CT, MR and selective angiogram of these patients were analyzed.

RESULTSOne week after treatment with embolization, the clinical symptoms of the 17 patients were remitted, and optic cacophony, nystagmus, exophthalmos and dropsy of conjunctiva disappeared. Two patients manifested surgical complications, one patient died. Sixteen patients survived. They were all followed up for more than 2 years, which showed one patient had handicap in movement, and in one patient the signs and symptoms of traumatic carotid-cavernous fistulas reoccurred 2 months after treatment.

CONCLUSIONSThe detachable balloon and/or coil embolization is safe and reliable. It is a good method to treat traumatic carotid-cavernous fistulas.

Adolescent ; Adult ; Angiography, Digital Subtraction ; Balloon Occlusion ; adverse effects ; instrumentation ; methods ; Carotid-Cavernous Sinus Fistula ; diagnosis ; mortality ; therapy ; China ; Embolization, Therapeutic ; adverse effects ; instrumentation ; methods ; Female ; Humans ; Injury Severity Score ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Prognosis ; Prospective Studies ; Risk Assessment ; Sampling Studies ; Survival Rate ; Tomography, X-Ray Computed ; Treatment Outcome ; Wounds, Penetrating ; complications

OBJECTIVETo establish a serum protein fingerprint model for prediction of liver metastasis from colorectal cancer by SELDI-TOF-MS analysis, and to determine the differentiatial proteins associated with the metastatic liver cancers.

METHODSData were collected from the Department of General Surgery in Zhongshan Hospital. A group of patients with colorectal cancer (CRC) without liver metastasis (n = 36) and another group with liver metastasis (n = 36) were included in this study. Serum samples were collected from peripheral venous blood before operation. Special serum protein or peptide fingerprint was determined by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). The obtained data were analyzed by Biomarker Wizard software to screen the serum protein markers discriminating colorectal cancer patients with and without liver metastasis. A serum protein fingerprint model was established. This model was blindly verified in of CRC patients with and 44 cases without liver metastasis.

RESULTSComparing the characteristic proteins in those two groups of patients, 10 specific protein peaks were identified with statistical significance (P < 0.05). According to m/z growing from small to large, they were: 2398, 2814, 4084, 4289, 4465, 6422, 6619, 11 482, 11 649 and 13 714. The predictive model had a sensitivity of 91.7% and a specificity of 97.2%. The validation showed a sensitivity of 75.0% and a specificity of 81.8%.

CONCLUSIONA predictive model based on differentiatial serum protein fingerprint with high sensitivity and specificity has been successfully established. It should be a very useful tool in detection and diagnosis of liver metastasis in colorectal cancer patients.

Aged ; Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Colorectal Neoplasms ; blood ; pathology ; Female ; Humans ; Liver Neoplasms ; blood ; diagnosis ; secondary ; Male ; Middle Aged ; Neoplasm Proteins ; blood ; Peptide Mapping ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

Matrix-assisted laser desorption/ionization time-of-flight imaging mass spectrometry (MALDI-TOF-IMS) can analysis unknown compounds in sections and obtain molecule imaging by scanning biological tissue sections,which has become a powerful tool for the research of biomarker,lipid distribution and drug metabolism,etc.This article reviews the application of this technique in protein identification,clinical application,drug discovery,lipid research and brain injury.

Objective To investigate the clinical characteristics of patients with advanced hepatic alveolar echinococcosis, so as to provide more evidence for the diagnosis and treatment of advanced hepatic alveolar echinococcosis. Methods The clinical records of 175 cases with hepatic alveolar echinococcosis admitted to the Affiliated Hospital of Qinghai University from 2012 to 2017 were retrospectively reviewed. Results Among the 175 cases with hepatic alveolar echinococcosis, the male to female ratio was 1∶1.6, and the mean age was (38.6 ± 12.9) years (range, 11 to 69 years). 93.7% of the patients (164 cases) were Tibetan, and 79.4% (139 cases) were from Guoluo Prefecture. 73.7% (129 cases) were herdsmen, and 58.3% (102 cases) were illiterate. The major clinical symptoms included upper abdominal pain, yellowing of skin and mucosa, and scleral icterus. 69.1% of the lesions were located in bilateral livers, and 63.4% of the cases have 2 and more lesions. 53.7% of the cases have their inferior vena cava involved by the lesions. In addition, the most affected neighboring organ was diaphragmatic muscle (21.7%), and lung was the most affected distant metastatic site (40.6%). Conclusions Advanced hepatic alveolar echinococcosis is highly prevalent in Guoluo Prefecture, Qinghai Province. Screening of this disease is required in this region and rational treatment scheme should be developed.

OBJECTIVES: To establish the imaging mass spectrometry for analysis of differentially expressed proteins distribution in the rat brains with diffuse axonal injury （DAI） based on matrix assisted laser desorption/ionization-time of flight imaging mass spectrometry （MALDI-TOF-IMS）. METHODS: MALDI-TOF-IMS scanning were conducted on the brains of DAI group and control group in the m/z range of 1 000 to 20 000 using AutoflexⅢ MALDI-TOF spectrometer. ClinProTool 2.2 software was used for statistical analysis on the data of two groups, and then the differentially expressed proteins were picked out to conduct imaging. The distribution of the proteins with different m/z in the rat brains was observed. RESULTS: Five proteins with different m/z, including 4 963, 5 634, 6 253, 6 714 and 7 532, differentially expressed in the rat brains with DAI. CONCLUSIONS MALDI-TOF-IMS can be used for studying the differentially expressed proteins in rat brains with DAI and the analysis method is established for exploring the distribution of differentially expressed proteins in the rat brains with DAI using imaging mass spectrometry.
Animals ; Brain/pathology* ; Diffuse Axonal Injury/pathology* ; Proteins/metabolism* ; Proteome/metabolism* ; Proteomics ; Rats ; Software ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

BACKGROUNDData on the epidemiology of hypertension in Chinese non-dialysis chronic kidney disease (CKD) patients are limited. The aim of the present study was to investigate the prevalence, awareness, treatment, and control of hypertension in the non-dialysis CKD patients through a nationwide, multicenter study in China.

METHODSThe survey was performed in 61 tertiary hospitals in 31 provinces, municipalities, and autonomous regions in China (except Hong Kong, Macao, and Taiwan). Trained physicians collected demographic and clinical data and measured blood pressure (BP) using a standardized protocol. Hypertension was defined as systolic BP ≥ 140 mmHg and/or diastolic BP ≥ 90 mmHg, and/or use of antihypertensive medications. BP < 140/90 mmHg and < 130/80 mmHg were used as the 2 thresholds of hypertension control. In multivariate logistic regression with adjustment for sex and age, we analyzed the association between CKD stages and uncontrolled hypertension in non-dialysis CKD patients.

RESULTSThe analysis included 8927 non-dialysis CKD patients. The prevalence, awareness, and treatment of hypertension in non-dialysis CKD patients were 67.3%, 85.8%, and 81.0%, respectively. Of hypertensive CKD patients, 33.1% and 14.1% had controlled BP to < 140/90 mmHg and < 130/80 mmHg, respectively. With successive CKD stages, the prevalence of hypertension in non-dialysis CKD patients increased, but the control of hypertension decreased (P < 0.001). When the threshold of BP < 130/80 mmHg was considered, the risk of uncontrolled hypertension in CKD 2, 3a, 3b, 4, and 5 stages increased 1.3, 1.4, 1.4, 2.5, and 4.0 times compared with CKD 1 stage, respectively (P < 0.05). Using the threshold of < 140/90 mmHg, the risk of uncontrolled hypertension increased in advanced stages (P < 0.05).

CONCLUSIONSThe prevalence of hypertension Chinese non-dialysis CKD patients was high, and the hypertension control was suboptimal. With successive CKD stages, the risk of uncontrolled hypertension increased.

Adult ; Aged ; Awareness ; Female ; Humans ; Hypertension ; complications ; epidemiology ; therapy ; Male ; Middle Aged ; Prevalence ; Renal Insufficiency, Chronic ; complications