To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.

Cyclin dependent kinases (cdks) play an important role in the pathogenesis of multiple neurodegenerative diseases. To explore the possibility of cdks-related gene therapy for neurodegenerative diseases, we packed recombinant adeno-associated virus (rAAV) encoding cdc2-siRNA. The expressing plasmid pAAV-MCS-EGFP-U6-cdc2-siRNA was constructed by using molecular biological techniques. The rAAV encoding cdc2-siRNA (rAAV-EGFP-U6-cdc2-siRNA) was packed by calcium phosphate mediated co-transfection of the plasmid pAAV-MCS-EGFP-U6-cdc2-siRNA, p-RC and p-Helper into AAV-293 cells. DNA sequencing proved the successful construction of U6-cdc2-siRNA in pAAV-MCS-EGFP. Seventy-two h after packaging, the expression of EGFP could be detected in AAV-293 cells. Western blotting revealed that cdc2 gene expression in AAV-293 cells was down-regulated markedly after transfection with rAAV-EGFP-U6-cdc2-siRNA, which evidenced the satisfactory silencing effect of this virus. It was concluded that the packaging of rAAV encoding cdc2-siRNA was successful. rAAV encoding cdc2-siRNA could silence cdc2 gene effectively, which might offer a novel means for the treatment of neurodegenerative diseases.

To study the relationship between the late postmortem interval (PMI) and trimethylamine-nitrogen (TMA-N) in postmortem tissues of cadaver, TMA-N in muscles, livers and kidneys of rats was measured at different postmortem intervals (PMI) by using a modified spectrophotometric method. The results indicated that the detection sensitivity of TMA-N was 1 mg/L, and there was a good linear correlation between the value of absorbance (A value) and TMA-N at the concentration of 1–10 mg/L (R² = 0.9991). Although TMA variation in muscles was different from that in inner organs during the time since death, TMA-N changes in cadaver tissues was positively correlated with PMI. During 2 to 7 d since death, the best correlation between PMI and TMA-N concentration was found in muscles. With PMI as an independent variable, the cubic polynomial regression equation was y= −0.457x³+6.519x²−24.574x+27.207 (R²=0.969). During 3 to 8 days since death, PMI was best correlated with TMA-N concentration in inner organs. With PMI as the independent variable, the cubic polynomial regression equation was y=0.509x³−9.153x²+55.727x−95.819 (R²=0.953). It was concluded that TMA-N in tissues could be used as a new estimator for late PMI. The method used in this study offered advantages such as accuracy, sensitivity, little samples required and wide PMI estimation.

The anti-cancer effects of betulinic acid (BA) on Jurkat cells and its in vitro mechanism were examined by using MTT assay. Apoptosis was detected by using Hoechst33258 staining and annexin-V/PI double-labeled cytometry. The effects of betulinic acid on the cell cycle of Jurkat cells were studied by propidium iodide method. RT-PCR and Western blotting were used to analyze the changes of cyclin D3, bcl-xl mRNA and protein levels in Jurkat cells after treatment with betulinic acid. Our results showed the proliferation of Jurkat cells was decreased in betulinic acid-treated group with a 24-h IC50 value being 70.00 μmol/L. Betulinic acid induced apoptosis of Jurkat cells in a time- and dose-dependent manner. The number of Jurkat cells treated with betulinic acid showed an increase in G₀/G₁ phase and decrease in S phase. After treatment with 0, 20, 60, 100 μmol/L betulinic acid for 24 h, the number of Jurkat cells was increased from (31.00±1.25)% to (58.84±0.32)% in G₀/G₁ phase, whereas it was decreased from (61.45±1.04)% to (35.82±1.95)% in S phase. PBMCs were less sensitive to the cytotoxicity of betulinic acid than Jurkat cells. The expressions of cyclin D3, bcl-xl mRNA and protein were decreased sharply in Jurkat cells treated with betulinic acid. It is concluded that betulinic acid is able to inhibit the proliferation of Jurkat cells by regulating the cell cycle, arrest cells at G₀/G₁ phase and induce the cell apoptosis. The anti-tumor effects of betulinic acid are related to the down-regulated expression of cyclin D3 and bcl-xl.

Current permanent right ventricular and right atrial endocardial pacing leads are implanted utilizing a central lumen stylet. Right ventricular apex pacing initiates an abnormal asynchronous electrical activation pattern, which results in asynchronous ventricular contraction and relaxation. When pacing from right atrial appendage, the conduction time between two atria will be prolonged, which results in heterogeneity for both depolarization and repolarization. Six patients with Class I indication for permanent pacing were implanted with either single chamber or dual chamber pacemaker. The SelectSecure 3830 4-French (Fr) lumenless lead and the SelectSite C304 8.5-Fr steerable catheter-sheath (Medtronic Inc., USA) were used. Pre-selected pacing sites included inter-atrial septum and right ventricular outflow tract, which were defined by ECG and fluoroscopic criteria. All the implanting procedures were successful without complication. Testing results (mean atrial pacing threshold: 0.87 V; mean P wave amplitude: 2.28 mV; mean ventricular pacing threshold: 0.53V; mean R wave amplitude: 8.75 mV) were satisfactory. It is concluded that implantation of a 4-Fr lumenless pacing lead by using a streerable catheter-sheath to achieve inter-atrial septum or right ventricular outflow tract pacing is safe and feasible.

This study investigated the changes of CD4⁺ CD25⁺ regulatory T cells (Tregs) in peripheral blood of patients with hepatocellular carcinoma before and after transcatheter arterial chemoembolization (TACE). The proportion of CD4⁺ CD25⁺ Tregs among CD4⁺ T lymphocytes in peripheral blood of 33 patients with hepatocellular carcinoma was determined by flow cytometry before, 1 week and 1 month after TACE. And 25 healthy volunteers served as control. One month after TACE, the patients were divided into two groups: 22 in group A, who were in stable condition or getting better; and 10 in group B, who were deteriorating. One patient died and was excluded. The results showed that the percentage of CD4⁺CD25⁺ Tregs among CD4⁺ T lymphocytes did not significantly change in the 33 patients 1 week after TACE as compared with that before TACE, however, the difference was significant (P<0.01) between the patients with hepatocellular carcinoma and the healthy subjects. The percentage of CD4⁺ CD25⁺ Tregs among CD4⁺ T lymphocytes in group A 1 month after TACE was decreased significantly in comparison with that before and 1 week after TACE (P<0.01), whereas, that in group B was increased significantly 1 month after TACE (P<0.01). It was concluded that patients with hepatocellular carcinoma had a higher proportion of CD4⁺CD25⁺ Tregs in peripheral blood. TACE did not significantly affect the level of CD4⁺ CD25⁺ Tregs within short time (such as 1 week). The proportion of CD4⁺CD25⁺ Tregs in peripheral blood 1 month after TACE was related to the prognosis of hepatocellular carcinoma.

In order to investigate the effect of vitamin A (VA) on the secretion of IFN-γ and IL-4 in Mycoplasma Pneumoniae (MP)-induced A549 cells, A549 cells were co-cultured with MP for different time lengths and then the levels of IFN-γ and IL-4 in the cell culture supernatants were detected before and after treatment with different concentrations of VA by using the enzyme-linked immunosorbent assay (ELISA). The results showed that the level of IFN-γ and IL-4 in the supernatants of MP-induced A549 cells was much higher than that in non-induced cells (P<0.01). After application of VA, IL-4 level was not increased until the concentration of VA was up to 0.5×10⁻⁵ mol/L (P<0.01). However, with concentration of VA increased up to 1×10⁻⁴ mol/L, IL-4 was significantly suppressed (P<0.01). It was concluded that MP could induce the secretion of IFN-γ and IL-4 in A549 cells. VA could inhibit the secretion of IFN-γ and increase the IL-4 level in MP-induced A549 cells. However, high concentration of VA had an inhibitory effect on the secretion of IL-4 as well as on the IFN-γ. These data provided a theoretical basis for the application of VA in MP pneumonia in the clinical practice.

This study investigated the association of apolipoprotein A5 (apoA5) gene polymorphism at position −1131T>C with cerebral infarction in patients with type 2 diabetes. A total of 256 type 2 diabetic patients without cerebral infarction (T2DM), 220 type 2 diabetic patients with cerebral infarction (T2DMCI) and 340 healthy subjects were recruited from the same region (Hubei province, China). The genotype of apoA5 −1131T〉C was analyzed by polymerase chain reaction, followed by restriction fragment length polymorphism (PCR-RFLP). Total cholesterol, HDL cholesterol, LDL-cholesterol and triglycerides were quantitatively detected by using standard enzymatic techniques. The results showed that the prevalence of the apoA5 −1131C allele was significantly higher in T2DMCI group than that in control group (42.7% versus 31.2%, P<0.01). The carriers of rare C allele had higher TG levels as compared with carriers of common allele in the three groups (P<0.01). Logistic regression models, which were adjusted for age, gender, blood pressure, BMI, FBS, smoking, LDL-C and HDL-C, revealed that patients carrying the apoA5 −1131C allele and CC homozygotes were at high risk for T2DMCI. It was concluded that the apoA5 −1131C allele variant is an independent genetic risk factor for T2DMCI.

To investigate the role of platelet membrane glycoprotein (GP) Ib/IX/V complex and its subunit GP Ibα in patients with hemorrhagic thrombopathy (HT), the expressions of GP Ib/IX/V complex and GP Ibα, defined as mean fluorescence intensity (MFI), were assessed by flow cytometry. The maximum aggregation of platelet was determined by turbidity method. These indicators were compared among 68 HT patients with the presenting complaint of hemorrhage, 33 well-controlled HT patients and 32 normal healthy subjects. The results showed that the MFI of GP Ib/IX/V complex and GP Ibα was markedly lower in HT patients with current hemorrhage than that in the healthy subjects, with difference being statistically significant (P<0.05). There was no significant difference in the expressions of GP Ib/IX/V complex and GP Ibα between well-controlled HT patients and normal healthy subjects (P>0.05). It was concluded that the expression of GP Ib/IX/V complex, the receptor of thrombin and von Willebrand factor, was down-regulated in HT patients with current hemorrhage, which might result in the dysfunction of platelet aggregation and recurrence of HT.

To study the effect of endotoxin on liver apoptosis, L02 liver cells were cultured and passaged in vitro, and then stimulated by endotoxin at 10 mg/mL for 4, 8, 16 and 24 h respectively. Liver apoptosis was flow cytometrically and fluorescently detected. Immunohistochemistry was used to detect the delivery of smac and caspase9. The delivery of liver cell smac and the activity of caspase3 were measured by caspase3 assay kit. The hepatic failure models of rats were established by using D-galactosamine. The blood serum and liver tissues were collected for the detection of the liver function, the level of endotoxin and the activity of caspase3 by using chromogenic substrate limulus amebocyte lysate method (LAL) and caspase3 active assay kit. The expression of smac and caspase9 in liver cells was detected by Western blotting. With in vitro study, the L02 cells stimulated by LPS condensed into conglobation and formed apoptotic bodies. After those cells were stained by hoechst, the apoptotic cells displayed blue color under the fluorescent microscope. The apoptosis rate was increased over time and the apoptosis was mainly of advanced stage. Meanwhile, the rate of smac delivery and activity of caspase9 and caspase3 were increased on L02 cell membrane. In vivo, hepatic failure and obvious endotoxemia were induced by injection of more than 200 mg/kg D-GalN. Hepatic mitochondria smac was reduced with dosage of D-GalN and, on the contrary, the activity of caspase3 was increased. D-GalN at 200 mg/kg increased Caspase9 while D-GalN at 300 mg/kg decreased caspase9. Mitochondria signal channel plays an important role in the endotoxin-induced apoptosis of hepatic cells by promoting the release of smac from mitochondria to cytoplasm and activating caspase9 and caspase3 in its low-level channel.

This study investigated the effect of RhoC GTPase on the proliferation and metastasis of cervical cancer cells, SiHa cells, in vitro. RhoC siRNA was introduced into SiHa cells to silence the RhoC gene. The mRNA and protein expression of RhoC, before and after RhoC siRNA transfection, was examined by RT-PCR and Western blotting, respectively. The proliferation and apoptosis of SiHa cells were examined by MTT assay and flow cytometry (FACS), respectively. Adhesive rate was evaluated by Matrigel adhesive assay, and the invasive capability and migration capability were assessed by transwell invasive assay and migration assay, respectively. The results showed that after the RhoC siRNA transfection, the mRNA and protein expression of RhoC was down-regulated in SiHa cells. The down-regulation of RhoC GTPase did not affect the cell proliferation and apoptosis (P>0.05), but it did suppress SiHa cells’ adhesion to matrigel (P<0.01), the invasive capability (P<0.01) and the migration capability (P<0.01). It was concluded that RhoC obviously promotes the adhesion, invasion and migration of SiHa cells in vitro, but not proliferation and apoptosis, suggesting that RhoC plays an important role in the progression in cervical cancer.

To examine the changes in number and function of endothelial progenitor cells (EPCs) from peripheral blood (PB) in hypertension disorder complicating pregnancy (HDCP), 20 women with HDCP and 20 normal pregnant women at the third trimester were studied. Mononuclear cells (MNCs) from PB were isolated by Ficoll density gradient centrifugation. EPCs were identified by positive expression of both CD34 and CD133 under fluorescence microscope and positive expression of factor VIII as shown by immunocytochemistry. The number of EPCs was flow-cytometrically determined. Proliferation and migration of EPCs were measured by MTT assay and modified Boyden chamber assay, respectively. The adhesion activity of EPCs was detected by counting the number of the adherent cells. The results showed that, compared with normal pregnant women, the number of EPCs was significantly reduced in HDCP (4.29%±1.21% vs 15.32%±2.00%, P<0.01), the functional activity of EPCs in HDCP, such as proliferation (13.45%±1.68% vs 18.45%±1.67%), migration (37.25±7.28 cells/field vs 67.10±9.55 cells/field) and adhesion activity (20.65±5.19 cells/field vs 34.40±6.72 cells/filed) was impaired (P<0.01). It is concluded that the number and function of EPCs are significantly decreased in HDCP.

To study the expression and implication of HIF-1α and VEGF-C in non-small cell lung cancer (NSCLC) and its relationship with clinical pathological features of NSCLC, immunohistochemical SP was used to detect the expression of HIF-1α and VEGF-C proteins in 48 NSCLC tissues and the same para-cancerous tissues. The positive rates of HIF-1α and VEGF-C were 70.8% (34/48) and 68.8% (33/48) respectively. The expression of HIF-1α protein was detected in a significantly greater proportion in NSCLC carcinoma tissues than that in para-cancerous tissues (12.5% and 16.7%, P<0.05). The positive rates of HIF-1α and VEGF-C were correlated with lymph node metastasis and TNM stage. No relationship was found between the two factors and age, sex, pathological subtypes and histological grades. The positive rates between HIF-1α and VEGF-C were correlated (P<0.05). HIF-1α and VEGF-C were over-expressed in NSCLC. They may be involved in the carcinogenesis of NSCLC, and play an important role in invasion and metastasis of NSCLC. HIF-1α and VEGF-C work synergically in the process of NSCLC.

In order to explore the possibility to predict the risk factors for postoperative complications and survival time, the clinical data of 152 patients (including 116 males and 36 females) who had undergone neo-adjuvant therapy and surgery for stage IIIA and B non-small cell lung cancer (NSCLC) were retrospectively analyzed. Demographic data, preoperative functional parameters, staging, induction regimen (chemotherapy alone or associated with radiotherapy), associated disorders, and data about operation were collected. Chi-square test and multivariate analysis fitting the unconditional logistic regression model were performed to identify predictors of postoperative complications, while Kaplan-Meier and multivariate Cox proportional hazard model were employed to identify predictors of survival time, respectively. The univariate analysis demonstrated that forced expiratory volume in 1 second predicted percent (FEV1%, P=0.040) and associated disorders (P=0.020) were the predictive factors of complications, but multivariate analysis found no independence factors (P>0.05) of it. Univariate Kaplan-Meier analysis showed that stage (P=0.050) and pneumonectomy (P=0.018) affected the survival time. However, multivariate Cox proportional hazard model analysis demonstrated that only pneumonectomy (P=0.026) was associated with a decreased survival time, but no differences between right and left pneumonectomy were found. The results suggest that the risk factor for postoperative complications is acceptable, and pneumonectomy is associated with increased mortality, which should be performed only in stage III NSCLC patients.

Presented in this study were three cases of lung cancer undergoing pulmonary venoplasty. In the 3 patients with central type of carcinoma of lung involving pulmonary vein, the main branch of right superior pulmonary vein and the distal end of the superior-lobe vein were occluded. The root part of the vein of right-middle lobe, plus part of vessel of of right superior vein was resected. The right superior vein was reconstructed by continuous 6-0 Prolene sutures. After the operation, the reconstructed was patent and the surgical margin was tumor-free. Postoperatively, clinical manifestations and plain chest films did not show any signs of venous blockade. The patients were discharged healed 3 weeks after the operation. The technical details of the surgery were presented, the improvements on the basis of traditional methods were discussed and its clinical application was evaluated. It is concluded that pulmonary venoplasty is a safe and feasible operation. Further improvement of the surgery will help conserve more lung tissue and benefit more patients because of expanded indications.

In this study, in vitro chemosensitivity testing was conducted on primary cultured breast cancer cells from 96 patients with breast cancer, and the results showed that the cells from a few patients with primary breast cancer developed multidrug resistance (MDR) prior to the first chemotherapy exposure. All the cells from the recurrent cancer patients had MDR. The findings suggested that patients having MDR would benefit from high-dose chemotherapy (HDC) regimens. In vitro chemosensitivity screening, which was aimed at improving the therapeutic efficacy and minimizing side effects, helps in choosing individualized treatment for breast cancer.

In this study, a recurrent massive phyllodes tumor of the breast was surgically removed and the grafting was used to repair the local skin defects. A 29-y female patient had recurring breast phyllodes tumor of extremely large size in the chest wall after the excision of a previous tumor. The massive phyllodes tumor was eliminated by completely removing the layers of the skin and tissues above the costal bones with incisal margin being 2 cm away from the tumor lesion. The latissimus dorsi myocutaneous flap, lateral thoracic skin flap, and rectus abdominis myocutaneous flap were grafted to replace the lost tissues due to the surgery. Anti-infection and anticoagulation treatments were used after the surgery. The graft flaps had sufficient blood supply and good blood circulation, and the incisions mostly healed. The outcome of the surgery was acceptable. For the surgical treatment of the massive phyllodes tumor in the chest wall, it is an alternative of choice to use latissimus dorsi myocutaneous flap, lateral thoracic skin flap and rectus abdominis myocutaneous flap in combination for skin grafting.

The association between the single nucleotide polymorphisms (SNPs) in −174G/C and −634C/G of interleukin-6 (IL-6) promoter region and prostate cancer was examined in the population of Han people in Hubei region. TaqMan PCR was employed for the gene-typing of −174G/C and −634C/G in promoter region of IL-6 gene to compare the prostate cancer patients and normal controls in terms of genotype frequency, allele frequency and risk of prostate cancer. Enzyme-linked immunosorbent assay (ELISA) was used for the detection of IL-6 concentration in peripheral blood of the patients with prostate cancer and the relationship between the IL-6 level and the genotype was studied. Our results showed that in all the subjects, the genotype of genetic locus −174G/C was found to be GG and no CG and CC were observed. There was a significant difference in gene frequency of GG, CG and CC of −634C/G and allele frequency of G and C between prostate cancer patients and normal controls (P<0.05) and the gene frequency of GG+CG increased with the clinical stages and pathological grades of prostate cancer. The IL-6 level in GG+CG group was significantly higher than that in CC group. It was concluded that no SNP in −174G/C IL-6 promoter region was found in the population of Han people in Hubei region. The SNP in −634C/G was, to some extent, associated with the development and progression of prostate cancer. The population with GG+CG genetype has higher risk for prostate cancer.

In order to investigate the inhibitory effect of matrine on the expression of prostate specific antigen (PSA) and androgen receptor (AR) in prostate cancer cell line LNCaP in vitro, LNCaP cells were treated with matrine at different concentrations (0.5, 1.0, 1.5, 2.0 g/L) for 12–36 h. The growth activities of cancer cells were determined by MTT colorimetric assay. The AR level was measured by Western blotting. The expression of PSA was detected by using AXSYM system-chemical luciferase methods. The results showed that matrine could effectively inhibit the growth of androgen-dependent prostate cancer cell line LNCaP in vitro in a time- and dose-dependent manner (P<0.05). It could obviously decrease the level of AR (P<0.01) and inhibit the expression of PSA in a dose-dependent manner (P<0.05) in LNCaP cells. It was concluded that matrine could significantly suppress the growth of LNCaP cells and inhibit the expression of PSA and AR of prostate cancer cells.

This study explored the possibility of using event-related potentials (ERP) for the measurement of picture-recognition memory and examined its correlation with the Chinese Wechsler Memory Scale-revised (WMS-RC) in patients with memory disorder caused by severe traumatic brain injury (sTBI). The subjects included 20 sTBI patients with memory disorder and 22 healthy individuals. Memory function was measured by using WMS-RC. Behavioral and ERP responses were recorded on-line during performance on a battery of picture recognition and the responses were analyzed off-line for recognition memory effects. Mean memory quotient (MQ) of patients with sTBI was significantly lower than that of the control group. Mean reaction time (RT) was significantly longer and the mean correctness rate (CR) of picture recognition was significantly lower in sTBI group than that of the controls. In controls, the main components of average ERP of picture recognition includes two positive-going waves, designated as P₁₇₀ and P₅₀₀, that appear 170 ms and 500 ms after stimulation when the subject could later successfully recall and recognize the pictures. P₅₀₀ amplitude of target stimulus was significantly higher than that of non-target stimulus. Compared to controls, P₅₀₀ responses of sTBI group were significantly delayed in latency (P<0.001) and lower in amplitude (P<0.001). P₅₀₀ latency showed significant negative correlation with MQ and the scores of “addition”, “visual recognition”, “picture recall”, “visual reproduction” and “tactile memory” in WMS-RC. ERP of picture recognition provides a neurophysiological approach to directly assess memory impairment, and P₅₀₀ may serve as a helpful index for memory disorder caused by sTBI in forensic practice.

This study observed the protective effect of hypercapnic acidosis preconditioning on rabbit heart suffered from ischemia-reperfusion injury. Hypercapnic acidosis was established in animals with mechanical hypoventilation before ischemia-reperfusion. Thirty-two rabbits were randomly divided into 4 groups, with each having 8 aminals in term of the degree of acidification: hypercapnic acidosis group A (group A), hypercapnic acidosis group B (group B), hypercapnic acidosis group C (group C), ischemia and reperfusion group (group IR). Animals in group IR were ventilated normally (tidal volume: 15 mL/kg, breathing rate 35 bpm). The PETCO₂ was maintained at the level of 40–50 mmHg for 30 min. Animals in groups A, B, C received low-frequency, low-volume ventilation to achieve hypercarbonic acidosis and the target levels of PETCO₂ were 75–85,65–75, 55–65 mmHg, respectively, with levels being maintained for 5 min. The animals then were ventilated normally to lower PETCO₂ to 40–50 mmHg. The left anterior branch artery of all the animals was ligated for 30 min and reperfused for 180 min. Then the infarct size was calculated. The cardiomyocytes were morphologically observed and ECG and hemodynamics were monitored on continuous basis. Acid-base balance was measured during procedure. Our results showed that the infarct size was (48.5±11.5)% of the risk area in the control group and (42.4±7.9)% in group C (P>0.05). Mean infarct size was significantly smaller in group B (34.5%±9.4%) (P<0.05 vs control group) and group A (31.0%±9.1%) (P<0.01 vs control group). It is concluded that HA-preconditioning can effectively protect the myocardium.

This study investigated the effects of propofol on the mRNA expression of Toll-like receptor-4 (TLR4) in BV-2 cells during mimic ischemia-reperfusion (I/R) injury in vitro. BV-2 cells, a mouse microglia line, were cultured and divided into 4 groups at random: control group (group C), ischemia/reperfusion group (group I/R), low-dose propofol (25 μmol/L) intervention group (group PF₂₅) and high-dose propofol (100 μmol/L) intervention group (group PF₁₀₀). The mRNA expression of TLR4 and NF-κB was measured by means of RT-PCR. TNF-α levels in the supernatants of BV-2 cells were detected by ELISA. The results showed that the mRNA expression of TLR4 and NF-κB was significantly higher in groups I/R, PF₂₅ and PF₁₀₀ than in group C (P<0.01). And the TNF-α level in the supernatants was elevated in groups I/R, PF₂₅ and PF₁₀₀ as compared with that in group C (P<0.01). After pre-treatment with propofol, the mRNA expressions of TLR4 and NF-κ B and the TNF-α level were significantly decreased in groups PF₂₅ and PF₁₀₀ in comparison to those in group I/R (P<0.01). And the decrease in those indicators was more significant in group PF₁₀₀ than in group PF₂₅ (P<0.01). It was concluded that propofol exerted brain-protecting effects during I/R injury by suppressing the mRNA expressions of TLR4 and NF-κB and deceasing the TNF-α level.

The remodeling process of synapses and neurotransmitter receptors of facial nucleus were observed. Models were set up by facial-facial anastomosis in rat. At post-surgery day (PSD) 0, 7, 21 and 60, synaptophysin (p38), NMDA receptor subunit 2A and AMPA receptor subunit 2 (GluR2) were observed by immunohistochemical method and semi-quantitative RT-PCR, respectively. Meanwhile, the synaptic structure of the facial motorneurons was observed under a transmission electron microscope (TEM). The intensity of p38 immunoreactivity was decreased, reaching the lowest value at PSD day 7, and then increased slightly at PSD 21. Ultrastructurally, the number of synapses in nucleus of the operational side decreased, which was consistent with the change in P38 immunoreactivity. NMDAR2A mRNA was down-regulated significantly in facial nucleus after the operation (P<0.05), whereas AMPAR2 mRNA levels remained unchanged (P>0.05). The synapses innervation and the expression of NMDAR2A and AMPAR2 mRNA in facial nucleus might be modified to suit for the new motor tasks following facial-facial anastomosis, and influenced facial nerve regeneration and recovery.

To investigate the value of ultrasound speckle tracking imaging (STI) in the assessment of the short-axis and long-axis systolic function of the left ventricle (LV) in patients with type 2 diabetes mellitus (DM), 100 subjects with normal ejection fraction were studied, including 41 patients with DM only (DM group), 22 patients with both DM and left ventricular hypertrophy (DH group), and 37 healthy subjects (control group). Left ventricle systolic function in the long axis defined as longitudinal strain, and that in the short axis defined as radial strain, apical and basal LV rotations, and LV twist were assessed respectively. The results showed that average peak strain in the long axis at basal, middle and apical levels, and global peak strain were significantly decreased in the patient groups when compared with the control group (P<0.001 for each). The parameters in DH group were significantly lower than those in DM group (P<0.01 for each). There were no significant differences in average radial peak strain in the short axis at different levels, and global peak strain among the three groups (P>0.05). Apical and basal LV rotations, and LV twist were greater in the patient groups than in the control group (P<0.01 for each). Basal LV rotation and LV twist were greater in DH group than those in DM group (P<0.01). It was concluded that STI may be used to identify early abnormalities in patients with type 2 DM that have normal left ventricular systolic function.

This study was aimed to evaluate the relationship between carotid atherosclerotic plaque stability and the clinical symptoms in patients with carotid atherosclerotic plaques by using contrast-enhanced ultrasonography. Fifty patients with carotid atherosclerotic plaques were enrolled and examined with contrast-enhanced ultrasonography. The correlation of contrast agent enhancement of the carotid atherosclerotic plaques and the clinical symptoms was analyzed. The results showed that among the 50 patients, plaques were enhanced in the 23 patients with obvious clinical symptoms. In 27 patients without apparent clinical symptoms, plaques were enhanced sparsely in 15 patients and not enhanced in 12 patients. It was suggested that contrast-enhanced ultrasonography could be used for the examination of the microcirculation in carotid atherosclerotic plaques on real-time basis and serve as a new noninvasive approach for the assessment of stability of carotid atherosclerotic plaques.

This study evaluated the application of quantitative tissue velocity imaging (QTVI) in assessing regional myocardial systolic and diastolic functions in dogs with acute subendocardial ischemia. Animal models of subendocardial ischemia were established by injecting microspheres (about 300 μm in diameter) into the proximal end of left circumflex coronary artery in 11 hybrid dogs through cannulation. Before and after embolization, two-dimensional echocardiography, QTVI and real-time myocardial contrast echocardiography (RT-MCE) via intravenous infusion of self-made microbubbles, were performed, respectively. The systolic segmental wall thickening and subendocardial myocardial longitudinal velocities of risk segments before and after embolization were compared by using paired t analysis. The regional myocardial video intensity versus contrast time could be fitted to an exponential function: y=A·(1-exp^−β·t), in which the product of A and β provides a measure of myocardial blood flow. RT-MCE showed that subendocardial normalized A·β was decreased markedly from 0.99±0.19 to 0.35±0.11 (P<0.05) in 28 left ventricular (LV) myocardial segments after embolization, including 6 basal and 9 middle segments of lateral wall (LW), 8 middle segments of posterior wall (PW) and 5 middle segments of inferior wall (IW). However, there was no statistically significant difference in subepicardial layer before and after embolization. Accordingly, the ratio of A·β of subendocardial myocardium to subepicardial myocardium in these segments was significantly decreased from 1.10±0.10 to 0.31±0.07 (P<0.05). Although the systolic wall thickening did not change 5 min after the embolization in these ischemic segments (29%±3% vs 31%±5%, P>0.05), the longitudinal peak systolic velocities (Vs) and early-diastolic peak velocities (Ve) recorded by QTVI were declined significantly (P<0.05). Moreover, the subendocardial velocity curves during isovolumic relaxation predominantly showed positive waves, whereas they mainly showed negative waves before the embolization. This study demonstrates that QTVI can more sensitively and accurately detect abnormal regional myocardial function and post-systolic systole caused by acute subendocardial ischemia.

Transthoracic echocardiographic characteristics of 17 cases of cardiac amyloidosis (CA), a rare disease in China, were analyzed in order to improve the understanding of the disease. Seventeen cases of biopsy-proven CA, admitted to Wuhan Union Hospital from June 1994 to September 2008 were retrospectively reviewed. Twenty normal volunteers served as control group. Left atrial and ventricular functions and mitral inflow velocity were measured by two-dimensional, and Doppler echocardiography, and tissue Doppler imaging (TDI)-derived peak systolic wall motion velocities (Sv), peak early diastolic wall motion velocities (Ev), and peak late diastolic wall motion (Av) were measured at the septum, lateral, inferior and anterior corners of mitral annulus from the apical 4- and 2 chamber views. Compared with the control group, the interventricular septal thickness (IVSd), the left ventricular posterior wall (LVPWd), right ventricular transverse diameter (RVTDd) near the end of diastole and the interauricular septum thickness (IASs), left atrial anteroposterior diameter (LAADs), right atrial transverse diameter (RATDs) near the end of systole were increased significantly (all P<0.05) and left ventricular ejection fraction (LVEF) decreased (P<0.05) in the CA group. Compared with the control group, Sv, Ev at each wall and Av at almost all walls were significantly decreased in the CA group. In the CA group, Myocardial echoes of interventricular septum and free wall of left ventricle were enhanced evidently and distributed unevenly. The echoes presented as ground glass-like images, with some spotty hyper echoes. Both atria were enlarged, and LVEF decreased, with diastolic function impaired, and mild-moderate hydropericardium found in the CA group. It was concluded that echocardiography was a relatively sensitive and highly specific non-invasive method for the diagnosis of CA.

This study examined the effect of different b values on diffusion-weighted MR imaging (DWI) of human prostate by using single-shot spin echo echo planar imaging (SE-EPI) sequences, observed the normal appearances and measured apparent diffusion coefficient (ADC) values in anatomical regions of normal prostate. Twenty-four healthy volunteers (mean age: 32 y) were studied by using a 1.5T system with a phased array surface multicoil. Two kinds of single-shot SE-EPI sequence were used to perform DWI in the prostate in volunteers, with five b values being 0, 30, 300, 500 to 1000 s/mm². The image quality with different imaging parameters was analyzed and the ADC values in anatomical regions of normal prostate were measured. DWI of prostate was successfully obtained in all volunteers. The images were of good quality, without artifacts containing pixels within the prostate. The contrast was good between the different anatomical regions of the prostatic gland, i.e., the peripheral zone (PZ), which exhibited higher signal intensity, and the central gland (CG). Signal intensity contrast was related to the magnitude of b values. The ADC values in PZ and CG were (1.27±0.22)×10⁻³ mm²/s and (1.01±0.17)×10⁻³ mm²/s, respectively. The ADC values were found to be significantly higher in PZ than in CG (P<0.05, paired t-test). Significant differences were found between the slice-selecting component and both the read-out and phase-encoding components of the ADC values. It is concluded that SE-EPI is a suitable DWI sequence for human prostate. The contrast between PZ and CG is good when b values are low, while the diffusion and ADC values are accurate when b values are high. ADC values are higher in PZ than in CG in normal prostate. Diffusional anisotropy is present in normal prostatic tissue.