This study was carried out to investigate the role of intrinsic neuroprotective mechanisms in the occurrence and development of vascular cognitive impairment (VCI) with the goal of providing a target for the treatment and prevention of VCI. Inpatients with proven cerebral infarction on cranial computed tomography (CT) were recruited as the ischemic cerebrovascular diseases (ICVD) group, and the patients with mixed stroke were excluded. In ICVD group, 12 patients were diagnosed as having VCI and served as VCI group. Inpatients undergoing surgical operation in our hospital were enrolled as control group. Double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) was employed to detect the levels of hypoxia-inducible factor 1-alpha (HIF-1α), vascular endothelial growth factor (VEGF), nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) in the cerebrospinal fluid of patients with ICVD. Associations between the levels of these factors and the Mini-Mental State Examination (MMSE) score were evaluated. In ICVD and VCI groups, the levels of HIF-1α and NGF in the cerebrospinal fluid were markedly lower than those in control group (P=0.037 and P=0.000; P=0.023 and P=0.005). In ICVD and VCI groups, the MMSE score was negatively related to VEGF level in the cerebrospinal fluid (r=-0.327, P=0.021; r=-0.585, P=0.046). In VCI group, HIF-1α level was correlated with NGF level (r=0.589, P=0.044). HIF-1α and NGF are involved in ischemic and hypoxic cerebral injury. The HIF signaling pathway plays an important role in intrinsic neuroprotection. Upregulation and maintenance of HIF-1α and NGF expression may attenuate VCI. Changes in VEGF levels are related to the occurrence and development of cognitive impairment.
Biomarkers ; cerebrospinal fluid ; Brain-Derived Neurotrophic Factor ; cerebrospinal fluid ; Cerebral Infarction ; cerebrospinal fluid ; complications ; diagnosis ; Cognition Disorders ; cerebrospinal fluid ; complications ; diagnosis ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; cerebrospinal fluid ; Male ; Middle Aged ; Nerve Growth Factor ; cerebrospinal fluid ; Reproducibility of Results ; Sensitivity and Specificity ; Vascular Endothelial Growth Factor A ; cerebrospinal fluid

Alzheimer's disease (AD) is an age-related, progressive neurodegenerative disorder that occurs gradually and results in memory, behavior, and personality changes. Abnormal sphingolipid metabolism was reported in AD previously. This study aimed to investigate whether sphK1 could exacerbate the accumulation of amyloid protein (Aβ) and sharpen the learning and memory ability of the animal model of AD using siRNA interference. An adenovirus vector expressing small interfering RNA (siRNA) against the sphK1 gene (sphK1-siRNA) was designed, and the effects of sphK1-siRNA on the APP/PS1 mouse four weeks after treatment with sphK1-siRNA hippocampal injection were examined. SphK1 protein expression was confirmed by using Western blotting and ceramide content coupled with S1P secretion was evaluated by enzyme-linked immunosorbent assay (ELISA). Aβ load was detected by immunohistochemical staining and ELISA. Morris water maze was adopted to test the learning and memory ability of the APP/PS1 mice. A significant difference in the expression of sphK1 protein and mRNA was observed between the siRNA group and the control group. Aβ load in transfected mice was accelerated in vivo, with significant aggravation of the learning and memory ability. The sphK1 gene modulation in the Aβ load and the learning and memory ability in the animal model of AD may be important for the treatment of AD.
Alzheimer Disease ; diagnosis ; physiopathology ; therapy ; Animals ; Disease Models, Animal ; Gene Silencing ; Genetic Therapy ; methods ; Learning Disorders ; diagnosis ; physiopathology ; therapy ; Mice ; Mice, Transgenic ; Microinjections ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; RNA, Small Interfering ; administration & dosage ; genetics ; therapeutic use ; Treatment Outcome

We developed a method that allows us to label nociceptive neurons innervating tooth-pulp in rat trigeminal ganglion neurons using a retrograde fluorescence-tracing method, to record ATP-activated current in freshly isolated fluorescence-labeled neurons and to conduct single cell immunohistochemical staining for P2X1 and P2X3 subunits in the same neuron. Three types of ATP-activated current in these neurons (F, I and S) were recorded. The cells exhibiting the type F current mainly showed positive staining for P2X3, but negative staining for P2X1. The results provide direct and convincing evidence at the level of single native nociceptive neurons for correlation of the characteristics of ATP-activated currents with their composition of P2X1 and P2X3 subunits and cell size. The results also suggest that the P2X3, but not P2X1, is the main subunit that mediates the fast ATP-activated current in nociceptive neurons.
Action Potentials ; physiology ; Adenosine Triphosphate ; metabolism ; Animals ; Dental Pulp ; innervation ; physiology ; Nociceptors ; cytology ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, Purinergic P2X1 ; metabolism ; Receptors, Purinergic P2X3 ; metabolism ; Tissue Distribution ; Trigeminal Nerve ; cytology ; metabolism

This study explored the cumulative live birth rate after three ovarian stimulation in vitro fertilization (IVF) cycles for poor ovarian responders according to the Bologna criteria. In this retrospective cohort study, 479 poor ovarian responders according to the Bologna criteria in the first ovarian stimulation IVF cycle between July 2006 and January 2012 in our IVF centre were included. The cumulative live birth rate was calculated by optimistic and pessimistic methods. The cumulative live birth rate after three ovarian stimulation IVF cycles for poor ovarian responders according to the Bologna criteria was 12.7%-20.5%. The three-cycle cumulative live birth rate was 18.5%-24.5%, 13.2%-27.4% and 8.6%-14.9% for poor responders aged ≤35 years, 36-39 years and ≥40 years, respectively. In conclusion, poor responders according to the Bologna criteria can receive an acceptable cumulative live birth rate after three ovarian stimulation IVF cycles, especially poor responders aged <40 years.
Adult ; China ; epidemiology ; Female ; Fertilization in Vitro ; statistics & numerical data ; Humans ; Infertility, Female ; epidemiology ; therapy ; Live Birth ; epidemiology ; Middle Aged ; Ovulation Induction ; statistics & numerical data ; Pregnancy ; Treatment Outcome ; Young Adult

This study aimed to identify biochemical predictors of adverse perinatal outcomes in intrahepatic cholestasis of pregnancy (ICP). A total of 106 ICP cases were analyzed retrospectively by the combination of receiver operating characteristic curve and binary logistic regression analysis. "Adverse perinatal outcomes" included spontaneous preterm labor, meconium-staining of amniotic fluid, stillbirth and Apgar score ≤7 at 1 or 5 min. Total bile acid (TBA) [AUC=0.658, 95%CI (0.536, 0.781), P=0.031] was a valuable predictor for adverse perinatal outcomes. The critical value of TBA above which adverse perinatal outcomes were observed was 40.15 μmol/L (Youden's index=0.3). Binary multivariate logistic regression analysis revealed that the risk of adverse perinatal outcomes increased when TBA ≥40.15 μmol/L [OR=3.792, 95%CI (1.226, 11.727), P=0.021]. It is concluded that the risk of adverse perinatal outcomes in ICP increases when maternal TBA ≥40.15 μmol/L.
Abortion, Induced ; Adult ; Bile Acids and Salts ; analysis ; Biomarkers ; metabolism ; China ; Cholestasis, Intrahepatic ; diagnosis ; metabolism ; Female ; Humans ; Pregnancy ; Pregnancy Complications ; diagnosis ; metabolism ; Prognosis ; Reproducibility of Results ; Risk Assessment ; Sensitivity and Specificity ; Stillbirth ; Thiobarbituric Acid Reactive Substances ; analysis ; Young Adult

This paper aimed to study the ability of baicalein to block human cytomegalovirus (HCMV) infection in extravillous cytotrophoblasts (EVT) and its effect on the vasoactive intestinal peptide (VIP) expression in HCMV-infected EVT in vitro. A human trophoblast cell line (HPT-8) was chosen in this study. HCMV with 100 TCID50 was added into culture medium to infect HPT-8 cells, and then HCMV pp65 antigen was assayed by immunofluorescence staining. The infection status was determined by virus titration. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect virus DNA load in the infected cells. The expression of VIP mRNA and protein in the infected cells was measured by qRT-PCR, immunocytochemistry and Western blotting. Concentration of VIP secreted in supernatants was determined by ELISA. Red-stained HCMV pp65 antigens were found in infected HPT-8 cells 48 h after infection. HCMV replicated in large quantity in infected HPT-8 cells 4 days after infection, reaching a peak at day 6 post-infection. After treatment with baicalein, virus DNA load in infected HPT-8 cells was decreased (P<0.05), and the levels of VIP mRNA and protein, and the concentration were raised to the normal (P>0.05). Our study suggested that baicalein exerts a positive effect on the VIP expression in HCMV-infected EVT at maternal-fetal interface.
Antiviral Agents ; pharmacology ; Cell Line ; Cytomegalovirus ; drug effects ; physiology ; Flavanones ; pharmacology ; Humans ; Trophoblasts ; cytology ; drug effects ; metabolism ; virology ; Vasoactive Intestinal Peptide ; metabolism ; Virus Inactivation ; drug effects

Preoperative planning of corrective osteotomy with traditional radiography has limitations in regards to determining the ideal osteotomy location and orientation in three-dimensional femoral deformities. Though a successful operation can be planned preoperatively, intraoperative contingencies might adhere to the procedural plan in the performance of operation. To efficiently perform a planned procedure, proposed is a design to implement three-dimensional reconstruction photography, based on computer-tomography (CT) scan. A custom-made guide was designed to navigate the osteotomy as planned, and additionally, a personalized intramedullary nail was used for fixation after osteotomy. Three-dimensional (3D) photography of deformed femur was established based on the CT dataset and transferred into 3D photography processing software for further planning. Osteotomy planes were designed and adjusted at deformity sites to correct the 3D deformities. The methodology of a custom-made osteotomy guide was introduced in femoral corrective osteotomy, for the first time, to navigate the operation as planned. After the virtual osteotomy and reduction of bone segments, the parameters of a custom-made intramedullary nail were measured for manufacturing. Findings Virtual operation in computer shows complete correction of the 3D deformity. The osteotomy guide, obtained by rapid-prototyping techniques, navigates mimicking surgery on rapid-prototyping model of the involved femur as planned. Internal fixation was achieved using the custom-made intramedullary nail. Interpretation three-dimensional visualization introduces an advantage in preoperative planning for corrective osteotomy of 3D femoral deformity, and the custom-made osteotomy guide is crucial to realize such a deliberate plan during the actual procedures. The internal fixator, such as an intramedullary nail, can be modified or personalized for fixation in unique cases.
Computer-Aided Design ; Equipment Failure Analysis ; Femur ; abnormalities ; diagnostic imaging ; surgery ; Fiducial Markers ; Humans ; Imaging, Three-Dimensional ; methods ; Osteotomy ; methods ; Reconstructive Surgical Procedures ; methods ; Surgery, Computer-Assisted ; methods ; Tomography, X-Ray Computed ; methods

The type I interferon and IFNAR play an important role in hepatitis B virus (HBV) infection and anti-HBV therapy. However, its mechanism of action is still poorly understood. To gain more insights into the role of type I interferon and type I interferon receptor (IFNAR) in HBV infection, we established an HBV persistent replication IFNAR knockout (IFNAR(-/-)) mouse model and preliminarily applied this model. At first, the progeny of IFNAR(-/-) mouse was reproduced. Then hydrodynamic injection with pAAV/HBV1.2 plasmid was conducted to establish the persistent HBV replication IFNAR(-/-) mouse model. At last, we applied this model to evaluate the effect of nucleoside analogues entecavir (ETV) on HBV replication. It was found that there was no difference in the serum HBsAg and HBeAg levels and HBcAg expression in the liver tissue between the ETV treated groups and normal saline (NS) treated group, but the serum HBV DNA levels were significantly suppressed 10, 25, 40 and 55 days after the ETV treatment [P=0.035, P=0.00, P=0.149 and P=0.084, IFNAR knockout (KO) control group vs. C57BL/6 ETV groups, respectively; P=0.081, P=0.001, P=0.243 and P=0.147, IFNAR KO control group vs. IFNAR KO ETV groups, respectively]. Interestingly, there was no difference in serum HBV DNA levels between the ETV treated IFNAR(-/-) and C57BL/6 mice. This result suggests that HBV suppression during ETV treatments doesn't depend on type I interferon and IFNAR. Collectively, persistent HBV replication IFNAR(-/-) mouse model that we established is a useful and convenient tool to detect the function of the type I interferon and IFNAR in HBV infection and anti-HBV treatments.
Animals ; Chronic Disease ; Disease Models, Animal ; Female ; Hepatitis B ; genetics ; virology ; Hepatitis B virus ; physiology ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Receptor, Interferon alpha-beta ; genetics ; metabolism ; Virus Replication ; genetics

This study aims to find good markers for predicting the prognosis of patients with esophageal squamous cell carcinoma (ESCC). Vasculogenic mimicry (VM) and the expression of hypoxia inducible factor-lα (HIF-lα)/E-cad protein in ESCC were investigated by immunostaining. The association between VM, HIF-lα/E-cad and clinicopathologic characteristics and 5-year-survival rate of patients with ESCC was analyzed. A total of 160 ESCC specimens were involved in this study and 28 specimens of normal esophageal mucosa served as controls. VM channels were identified in 78 (48.75%) of the 160 ESCC specimens and none of the normal esophageal mucosa was found to have VM. The rates of high-expression of HIF-lα and E-cad in ESCC were 43.75% and 38.75%, while the rates in control were 17.86% and 71.43%, respectively (P<0.05 for all). VM and the expression levels of HIF-lα and E-cad were significantly related to lymph node metastasis, serosa infiltration, PTNM staging and 5-year-survival rates of patients with ESCC (P<0.05 for all). VM was positively correlated with HIF-lα but negatively with E-cad, and HIF-lα was negatively correlated with E-cad (P<0.001 for all). The 5-year-survival rate of patients with ESCC was 6.41% (5/78) in VM group and 65% (52/82) in non-VM group, 7.14% (5/70) in high HIF-lα expression group and 57.78% (52/90) in low HIF-lα expression group. Oppositely, the 5-year-survival rate in high E-cad expression group was 80.65% (50/62) and that in low E-cad expression group was 7.37% (7/98) (P<0.05 for all). Cox multifactor regression analysis indicated that lymph node metastasis, PTNM stage, VM and expression levels of HIF-lα and E-cad were independent risk factors of patients with ESCC (P<0.05 for all). Combined detection of VM, HIF-lα and E-cad plays an important role in predicting the invasion, metastasis and prognosis of patients with ESCC.
Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; metabolism ; Cadherins ; metabolism ; Carcinoma, Squamous Cell ; diagnosis ; metabolism ; Esophageal Neoplasms ; diagnosis ; metabolism ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Middle Aged ; Neovascularization, Pathologic ; diagnosis ; metabolism ; Prognosis ; Reproducibility of Results ; Sensitivity and Specificity ; Survival Rate

The specimens of ductal carcinoma in situ (DCIS) with early invasion, and specimens collected by core needle biopsy (CNB) tend to contain limited amount of invasive component, so it is imperative to explore a new technique which can assess HER2 gene status accurately for the limited invasive cancer component in these specimens. Dual staining technique of combining immunohistochemistry (IHC) for myoepithelial cells and single or dual probe chromogenic in situ hybridization (CISH) for HER2 gene was performed on routinely processed paraffin sections from 20 cases diagnosed as having DCIS with invasive cancer. Among them, 10 had fluorescence in situ hybridization (FISH)-confirmed amplification of HER2 and 10 had FISH-confirmed non-amplification of HER2. We successfully detected HER2 genetic signals and myoepithelial IHC markers (SMM-HC or CK5/6) simultaneously on a single section in all 20 specimens. Myoepithelial markers and HER2 signals detected by dual staining assay were consistent with those by individual technique performed alone. HER2 gene amplification results determined by dual staining assay were 100% consistent with those of FISH. Dual staining technique which allows simultaneous detection of myoepithelial marker protein and cancerous HER2 gene is feasible, and it has potential to be used in clinical practice for effective determination of HER2 amplification in limited invasive component.
Biomarkers, Tumor ; metabolism ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Chromogenic Compounds ; Female ; Gene Expression Profiling ; methods ; Humans ; Immunohistochemistry ; methods ; In Situ Hybridization, Fluorescence ; methods ; Neoplasm Invasiveness ; pathology ; physiopathology ; Receptor, ErbB-2 ; genetics ; metabolism ; Reproducibility of Results ; Sensitivity and Specificity