Objective To investigate the clinical value of serum homocysteine (Hcy),Cystatin C (CysC) detection in coronary heart disease prevention,diagnosis and treatment.Methods A total of 200 cases of coronary heart disease patients including 50 cases of acute myocardiac infarction (AMI) (AMI group),85 cases of unstable angina pectoris (UAP)(UAP group.) and 65 cases of stable angina pectoris (SAP) (SAP group) were selected,and 120 cases of healthy controls were selected as control group.The serum levels of Hcy and CysC were detected and compared.Results The serum levels of Hcy and CysC in AMI group,UAP group and SAP group were significandy higher than those in control group[Hcy:(22.35 ± 8.18),(16.54±7.56),(14.52±6.38) μmol/Lvs.(9.35±3.23) μmol/L; CysC:(1.32±0.27),(1.88± 0.66),(1.19 ± 0.46) mg/L vs.(0.80 ± 0.33) mg/L],and there were significant differences between two groups (P＜ 0.01).The serum level of Hcy in AMI group was higher than that in UAP group and SAP group [(22.35 ± 8.18) μ mol/L vs.(16.54 ± 7.56),(14.52 ± 6.38) μ mol/L],and there was significant difference (P ＜ 0.01).Conclusions The relationship between Hcy,CysC level and coronary heart disease is close,and Hcy levels increase with increasing degree of coronary heart disease.Detection of Hcy and CysC has some clinical value on prevention,diagnosis,treatment of coronary heart disease.

To study on the effects of Achyranthes bidentata on Tongsaimai pellets main active ingredients chlorogenic acid, isoliquiritin, harpagoside and glycyrrhizin in rats in vivo pharmacokinetic behaviors, a method for the simultaneous determination of chlorogenic acid, isoliquiritin, harpagoside and liquiritigenin in rat plasma was established by UPLC-MS/MS. The analysis was performed on a waters Acquity BEH C18 column (2.1 mm x 100 mm, 1.7 microm) with the mixture of acetonitrile and 0.1% formic acid/water as mobile phase, and the gradient elution at a flow rate of 0.3 mL x min(-1). The analytes were detected by tandem mass spectrometry with the electrospray ionization (ESI) source and in the multiple reaction monitoring (MRM) mode. It turned out that the analytes of Tongsaimai pellets groups C(max) and AUC(Q-infinity) values were higher than that with A. bidentata group, and the C(max) values of chlorogenic acid had significantly difference (P < 0.05), the AUC(0-infinity) values of chlorogenic acid and glycyrrhizin had significantly difference (P < 0.05); The T(max) and CL values of two groups had no significantly difference. Results showed that the established method was specific, rapid, accurate and sensitive for the studies of Tongsaimai pellets four main active ingredients in rat in vivo pharmacokinetic, and A. bidentata have varying degrees of effects on Tongsaimai pellets four main active ingredients in rat in vivo pharmacokinetic behaviors.
Achyranthes ; chemistry ; Animals ; Chalcone ; administration & dosage ; analogs & derivatives ; blood ; pharmacokinetics ; Chlorogenic Acid ; administration & dosage ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Glycosides ; administration & dosage ; blood ; pharmacokinetics ; Glycyrrhizic Acid ; administration & dosage ; pharmacokinetics ; Herb-Drug Interactions ; Male ; Pyrans ; administration & dosage ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

Objective To determine the feasibility of magnetically labeling and tracking mesenchymal stem cells(MSCs)in vitro by using a gadolinium and fluorescent bi-functionally transfection agent of polyethylenimine.Methods A gadolinium bifunctional transfection reagent complex was obtained after the linear polyethylenimine derivative(JetPEI-FluoR)was incubated with Gd-DTPA.Mesenchymal stem cells isolated from the bone marrows of SD rats were cultured and expanded.The mesenchymal stem cells were incubated with the bi-functional labeling agents.After labeling,the MSCs were examined with fluoroscope and electron microscope and the biological characters were detected including trypan blue exclusion test,MTT,and apoptosis detection.On a 1.5 T MR system,the labeled MSCs were examined with spin echo T1 WI and T2 WI and T1 measurement with mixed sequence.After labeling,the cells were cultured and undergone routine passage.Prior MR examinations were repeated for each passage of labeled cells.All data was statistically prolessed with SPSS for Windows.Results Of 5×105 MSCs incubated with the bi-functional agents,4.25×105 MSCs were successfully labeled,the percentage of labeled MSCs was 85% fluoroscopically.The high density electron particles of gadolinium observed electron microscopically existed around cellular apparatuses,especially around Golgi apparatus.In trypan blue exclusion test,the exclusion rate of labeled MSCs with incubation duration of 3,6,12,24 h was(96.55±2.90)%,(94.17±2.56)%,(97.16±3.12)% and(94.23±2.67)%,respectively.The corresponding exclusion rate of unlabeled MSCs was(95.86±2.67)%,(92.04±2.21)%,(93.38±3.64)%and(92.12±2.53)%,respectively.There was no statistical difference of trypan blue exclusion rate between labeled cells and control unlabeled cells within 24 hours of incubation(F=4.523,P＞0.05).In the proliferation test,the optical absorption value of labeled MSC with 2.5,5.0,10.0,20.0,30.0 and 40.0 μl bi-functional labeling agent was(0.1884±0.0151),(0.1878±0.0190),(0.1741±0.0160),(0.1135±0.0215),(0.1079±0.0145)and(0.0811±0.0079),respectively.The corresponding optical absorption value of unlabeled MSCs was(0.1940±0.0116).The optical absorption value of labeled cells was not affected in case of less than 30.0 μl of Gd-DTPA(q'=0.2225-0.9458,P＞0.05).The apoptosis index for labeled cells and unlabeled cells were 5.08% and 3.86%,respectively.On T1 WI,the signal intensity and T1 relaxation time of unlabeled cells and labeled cells were 240.3±24.7 and(2457±56)ms,336.2±20.7 and(1102±64)ms,respectively,and there were significant statistical difference(t=12.656,17.889,P＜0.01).The minimal amount of cells which was detectable for T1 WI was 5×103.After routine passage,the gadolinium in the cells gradually decreased and could be tracked by MRI until the fifth passage.Conclusions The gadolinium and fluorescent bi-functionally labeling rat bone marrow mesenchymal stem cell by using the transfection agent of polyethylenimine is feasible,efficient and safe.The labeled cells could be tracked in vitro on MR imaging.

ObjectiveTo discusse the relationship between static balance in laboratory approaches and dynamic balance in clinical assessment and identify the value of static and dynamic balance at functional outcome in hemiparetic stroke patients. MethodsNineteen stroke subjects were assessed in this study. The static balance was measured by postural sway test, the dynamic balance was measured by Berg balance scale(BBS)and Time up to go test(TUGT),the outcome was measured by FIM and 10m maximum walking speed(MWS). The level of association between the parameters of postural sway test and clinical variables were examined with Pearson's correlation coefficients. ResultsThe parameters of postural sway test was significantly negative related to BBS(r=-0.705--0.475,P<0.05);The parameters of postural sway close-eye test was significantly positive related to TUGT(r=0.508-0.583,P<0.05);The parameters of postural sway test was no related to FIM and MWS (r=-0.048--0.296;r=-0.404--0.01,P>0.05);BBS was significantly positive related to FIM and MWS(r=0.752;r=0.700,P<0.001). TUGT was significantly negative related to FIM and MWS(r=-0.600,P<0.01;r=-0.817,P<0.001).ConclusionClinical and laboratory balance assessments are related and that dynamic rather than static balance measures are valid indicators of functional outcome performance in hemiparetic stroke patients.

OBJECTIVETo evaluate clinical effects of pedicle fixation without bone fusion in treating thoracolumbar fractures through paraspinal approach.

METHODSFrom January 2006 to January 2009, 25 patients (15 males and 10 females) with thoracolumbar fractures were treated. The average age was 39.3 years,ranged from 17 to 49 years. According to classification, flexion fracture in 7 cases, brust fracture in 18 cases. There were no nervous injury, and radiology information showed the angle of sagittal vertebral body >20 degrees or collapse of vertebral body >40%,without vertebral injury. The operation were performed at 3 to 7 days after injury (mean 5 day). Internal fixation implants were removed at 8 to 12 months after operation. The height, kyphosis angle were measured before operation, 1 week and 24 months after operation,and Oswestry disability index (ODI) were compared before and after operation.

RESULTSAll patients were followed up for 24 months. Among them, 1 case was followed up at 30 months after operation. The operation time ranged from 70 to 110 (mean 90) minutes, the blood loss was 120 to 280 (mean 200) ml. The height of vertebral body and kyphosis angle were obviously corrected, and had significant differences between postoperation immediately and at the final follow-up (P<0.05). There were no differences after remove of internal fixation (P>0.05). The final ODI was (5.36 +/- 1.21)%, had statistical differences compared with preoperation (P<0.05).

CONCLUSIONFor flexion and burst thoracolumbar fractures without nervous injury, pedicle fixation without bone fusion is a good method,which has advantages of minimally invasive, rapid recovery, and maintain spinal motion segment.

Adolescent ; Adult ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; diagnostic imaging ; injuries ; surgery ; Male ; Middle Aged ; Spinal Fractures ; diagnostic imaging ; surgery ; Spinal Fusion ; methods ; Thoracic Vertebrae ; diagnostic imaging ; injuries ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

OBJECTIVETo assess the clinical effect and safety of the Chinese medicine Longbishu Capsule combined with mesylate doxazosin in the treatment of benign prostatic hyperplasia (BPH) of the kidney deficiency and blood stagnation type.

METHODSThis was a randomized, double-blind, double-simulation control study. We equally assigned 60 men diagnosed with BPH of the kidney deficiency and blood stagnation type to an experimental and a control group, the former treated with mesylate doxazosin plus Longbishu Capsule and the latter with mesylate doxazosin plus placebo. We compared the International Prostate Symptom Score (IPSS), quality of life (QOL), Chinese symptom score (CSS), maximal urinary flow rate (Qmax), and prostate volume between the two groups of patients before and after 6 months of medication.

RESULTSAfter treatment, there were 5 cured cases, 13 markedly effective cases, 9 effective cases, 1 ineffective case, and 2 eliminated cases in the experimental group, as compared with 2 cured cases, 8 markedly effective cases, 10 effective cases, 7 ineffective cases, and 3 eliminated cases in the control group. The total effectiveness rate was obviously higher in the former (96.4%) than in the latter (74.1%). IPSS, Qmax, and CSS were improved in both of the groups after medication, even more significantly in the experimental than in the control group (IPSS: 15.22 ± 2.98 vs 18.15 ± 5.88, P <0.05; Qmax: [13.56 ± 2.26] ml/s vs [11.78 ± 2.97] ml/s, P <0.05; CSS: 6.18 ± 2.13 vs 9.52 ± 3.15, P <0.05). Because of the difference in the QOL score between the two groups at the baseline (P = 0.038 <0.05), no more comparison was made in this aspect after treatment.

CONCLUSIONThe combination of Longbishu Capsule with mesylate doxazosin is safe and effective for the treatment of BPH.

Adrenergic alpha-Antagonists ; therapeutic use ; Capsules ; Double-Blind Method ; Doxazosin ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Prostatic Hyperplasia ; drug therapy ; physiopathology ; Quality of Life ; Treatment Outcome ; Urination

OBJECTIVETo study the expression of NFkappaB p65 and its target genes in intestinal metaplasia (IM), dysplasia (Dys), gastric cancer (GC) infected with Helicobacter pylori (Hp) and explore the mechanism of infection by cytotoxin-associated antigen A expressing Hp (CagA(+)Hp) in the development of gastric cancer.

METHODSCagA antibody in blood sample of 289 patients was determined by ELISA. Hp was detected by rapid urease test and Warthin starry staining. Expression of NFkappaB p65 and its target genes in IM, Dys and GC was examined by immunohistochemistry.

RESULTSIn IMI approximately II, IMIII, DysI, DysII approximately III and GC, the expression of NFkappaB p65 was significantly higher in patients with CagA(+)Hp infection than those without CagA Hp infection. In IMIII and DysII approximately III, the expression of NFkappaB p65, c-myc, CyclinD(1) and bcl-xl was significantly higher in patients with CagA Hp infection than those without CagA Hp infection. In gastric cancer infected with CagA(+)Hp, the expression of NFkappaB p65, c-myc, CyclinD(1) and bcl-xl was significantly higher in intestinal type than in diffuse type.

CONCLUSIONThere are different mechanisms in intestinal type and diffuse type in the development of gastric cancer. The occurrence of intestinal type gastric cancer is associated with CagA(+)Hp infection which by NFkappaB p65 upregulating the expression of c-myc, CyclinD(1),bcl-xl in patients with IMIII, DysII approximately III. It may be an effective method to prevent gastric cancer by inhibiting NFkappaB p65.

Adult ; Aged ; Antigens, Bacterial ; analysis ; Bacterial Proteins ; analysis ; Cyclin D1 ; metabolism ; Female ; Helicobacter Infections ; complications ; metabolism ; microbiology ; Helicobacter pylori ; Humans ; Male ; Middle Aged ; Precancerous Conditions ; metabolism ; microbiology ; pathology ; Proto-Oncogene Proteins c-myc ; metabolism ; Stomach Neoplasms ; metabolism ; microbiology ; pathology ; Transcription Factor RelA ; genetics ; metabolism ; bcl-X Protein ; metabolism

DNA mismatch repair gene mutS (2.56 kb) was PCR modified and cloned into a secretive prokaryotic expression vector pET32a (+) which carries a N-terminal His.tag + and thioredoxin sequence. MutS protein was expressed with high level after IPTG induction using the strain E. coli AD494(DE3). SDS-PAGE revealed that the expected protein with a molecular weight of 108 kD which is about 35% of the total bacterial proteins is almost soluble. The expected protein was purified directly by immobilized metal (Ni2+) chelation affinity chromatography and the purity is over 90%. MutS protein activity verified using mismatch DNA showed that the expression product can recognize and bind to base-pair mismatch specifically.
Adenosine Triphosphatases ; biosynthesis ; genetics ; isolation & purification ; Bacterial Proteins ; Base Pair Mismatch ; Chromatography, Affinity ; DNA ; metabolism ; DNA Repair ; DNA-Binding Proteins ; Escherichia coli Proteins ; biosynthesis ; genetics ; isolation & purification ; Magnesium ; pharmacology ; Molecular Weight ; MutS DNA Mismatch-Binding Protein ; Recombinant Proteins ; biosynthesis

OBJECTIVETo observe the effect of Cordyceps sinensis (CS) powder on renal oxidative stress and mitochondria functions in 5/6 nephrectomized rats, and to primarily explore its possible mechanisms.

METHODSTotally 30 male Sprague-Dawley rats were divided into the sham-operation group, the model group, and the treatment group by random digit table, 10 in each group. A chronic kidney disease (CKD) rat model was prepared by one step 5/6 nephrectomy. Rats in the treatment group were intragastrically administered with CS powder solution at the daily dose of 2 g/kg, once per day. Equal volume of double distilled water was intragastrically administered to rats in the sham-operation group and the model group. All medication lasted for 12 weeks. The general condition of rats, their body weight, blood pressure, 24 h proteinuria, urinary N-acetyl-β-D-glucosaminidase (NAG), serum creatinine (SCr) , and blood urea nitrogen (BUN) were assessed before surgery, at week 2, 4, 6, 8, 10, and 10 after surgery. Pathological changes of renal tissues were observed under light microscope. Morphological changes of mitochondria in renal tubular epithelial cells were observed under transmission electron microscope. Activities of antioxidant enzymes including reduced glutathione (GSH), manganese superoxide dismutase (MnSOD), and malondialdehyde (MDA) in fresh renal tissue homogenate were detected. Mitochondria of renal tissues were extracted to detect levels of mitochondrial membrane potential and changes of reactive oxygen species (ROS). And expressions of cytochrome-C (Cyto-C) and prohibitin in both mitochondria and cytoplasm of the renal cortex were also measured by Western blot.

RESULTS(1) Compared with the sham-operation group, body weight was significantly decreased at week 2 (P <0. 01), but blood pressure increased at week 4 (P <0. 05) in the model group. Compared with the model group, body weight was significantly increased at week 12 (P <0. 01), but blood pressure decreased at week 8 (P < 0. 01) in the treatment group. (2) Compared with the sham-operation group, 24 h proteinuria, urinary NAG, blood SCr and BUN significantly increased in the model group (all P <0. 01). Compared with the model group, blood and urinary biochemical indices all significantly decreased in the treatment group (all P <0. 01). (3) Results of pathological renal scoring: Glomerular sclerosis index, scoring for tubulointerstitial fibrosis, degree of tubulointerstitial inflammatory infiltration were all obviously higher in the model group than in the sham-operation group (all P <0. 01). All the aforesaid indices were more obviously improved in the treatment group than in the model group (all P <0. 01). (4) Compared with the sham-operation group, activities of MnSOD and GSH-Px were significantly reduced, but MDA contents obviously increased in the renal cortex of the model group (all P <0. 01). Compared with the model group, activities of MnSOD and GSH-Px obviously increased (P <0. 05, P <0. 01), but MDA contents obviously decreased in the renal cortex of the treatment group (P <0. 01). (5) Compared with the sham-operation group, the mitochondrial membrane potential significantly decreased, but ROS levels significantly increased in the model group (all P <0.01). Compared with the model group, mitochondrial transmembrane potential increased in the treatment group, thereby inhibiting the tendency of increased production of ROS (both P < 0. 01). (6) Results of Western blot showed that, compared with the sham-operation group, expression levels of mitochondrial Cyto-C and Prohibitin were significantly reduced in the renal cortex (P <0. 01), but significantly elevated in the cytoplasm of the model group (P <0. 01). Compared with the model group, each index was obviously improved in the treatment group with statistical difference (P <0. 05, P <0. 01).

CONCLUSIONCS powder had renal protection, and its mechanism might partially depend on in- hibition of oxidative stress and protection for mitochondria.

Acetylglucosaminidase ; metabolism ; Animals ; Blood Urea Nitrogen ; Cordyceps ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; Kidney Cortex ; Kidney Diseases ; Kidney Function Tests ; Male ; Malondialdehyde ; metabolism ; Mitochondria ; Nephrectomy ; Oxidative Stress ; drug effects ; Proteinuria ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

Objective: Patients with a history of ischemic stroke are at risk for a second ischemic stroke. This study aimed to investigate the relationship between carotid plaque enhancement on perfluorobutane microbubble contrast-enhanced ultrasonography (CEUS) and future recurrent stroke, and to determine whether plaque enhancement can contribute to risk assessment for recurrent stroke compared with the Essen Stroke Risk Score (ESRS). Materials and Methods: This prospective study screened 151 patients with recent ischemic stroke and carotid atherosclerotic plaques at our hospital between August 2020 and December 2020. A total of 149 eligible patients underwent carotid CEUS, and 130 patients who were followed up for 15–27 months or until stroke recurrence were analyzed. Plaque enhancement on CEUS was investigated as a possible risk factor for stroke recurrence and as a possible adjunct to ESRS. Results: During follow-up, 25 patients (19.2%) experienced recurrent stroke. Patients with plaque enhancement on CEUS had an increased risk of stroke recurrence events (22/73, 30.1%) compared to those without plaque enhancement (3/57, 5.3%), with an adjusted hazard ratio (HR) of 38.264 (95% confidence interval [CI]:14.975–97.767; P < 0.001) according to a multivariable Cox proportional hazards model analysis, indicating that the presence of carotid plaque enhancement was a significant independent predictor of recurrent stroke. When plaque enhancement was added to the ESRS, the HR for stroke recurrence in the high-risk group compared to that in the low-risk group (2.188; 95% CI, 0.025–3.388) was greater than that of the ESRS alone (1.706; 95% CI, 0.810–9.014). A net of 32.0% of the recurrence group was reclassified upward appropriately by the addition of plaque enhancement to the ESRS. Conclusion Carotid plaque enhancement was a significant and independent predictor of stroke recurrence in patients with ischemic stroke. Furthermore, the addition of plaque enhancement improved the risk stratification capability of the ESRS.