Objective:To investigate whether astragaloside IV (AS-IV) exerts neuroprotective effects via zinc ion (Zn 2+) modulation of mitochondria-associated endoplasmic reticulum membranes (MAMs) and to elucidate the possible mechanisms. Methods:PC12 cells (rat adrenal pheochromocytoma cells) were routinely cultured; they were divided into 7 groups: control group (routinely cultured), 2-DG group (treated with 2-DG at 50 μmol/L for 30 min), astragaloside IV+2-DG group (treated with 50 μmol/L astragaloside IV for 20 min, and then treated with 50 μmol/L 2-DG for 30 min), astragaloside IV group (treated with 50 μmol/L astragaloside IV for 20 min), N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (a zinc chelator, TPEN)+astragaloside IV+2-DG group (treated with 10 μmol/L TPEN for 10 min, 50 μmol/L astragaloside IV for 20 min, and then 50 μmol/L 2-DG for 30 min); TPEN group (treated with 10 μmol/L TPEN for 10 min), TPEN+2-DG group (treated with 10 μmol/L TPEN for 10 min and then treated with 50 μmol/L 2-DG for 30 min). The expressions of glucose-regulated protein (GRP)78, GRP94, cysteine-containing aspartate proteolytic enzyme caspases-8, B cell receptor associated protein 31 (BAP31) and mitochondrial fission protein 1 (Fis1) were detected by Western blotting. Annexin V-FITC/PI kit was used to detect the apoptosis level. Immunofluorescence was used to detect the Fis1 protein expression, and mitochondrial fluorescence probe TMRE was used to detect the opening of mitochondrial permeability transition pore (mPTP).Results:(1) Compared with the control group, the 2-DG group had significantly increased expressions of GRP78, GRP94, Caspase-8, Bap31 and Fis1, and green fluorescent intensity of Annexin V-FITC ( P<0.05); compared with the 2-DG group, the astragaloside IV+2-DG group had significantly decreased expressions of GRP78, GRP94, Caspase-8, Bap31 and Fis1, and green fluorescent intensity of Annexin V-FITC ( P<0.05); compared with the astragaloside IV+2-DG group, the TPEN+astragaloside IV+2-DG group had significantly increased expressions of GRP78, GRP94, Caspase-8, Bap31 and Fis1, and green fluorescent intensity of Annexin V-FITC ( P<0.05). (2) Compared with the control group, the 2-DG group had significantly enhanced Fis1 protein fluorescent intensity ( P<0.05); the astragaloside IV+2-DG group had significantly decreased Fis1 protein fluorescent intensity compared with 2-DG group ( P<0.05); Compared with astragaloside IV+2-DG group, TPEN+astragaloside IV+2-DG group had significantly enhanced Fis1 protein fluorescence intensity ( P<0.05); compared with the control group, the TPEN+2-DG group had significantly enhanced Fis1 protein fluorescent intensity ( P<0.05). (3) TMRE fluorescence intensity in 2-DG group was significantly decreased compared with control group ( P<0.05); TMRE fluorescence intensity in astragaloside IV+2-DG group was significantly enhanced compared with 2-DG group ( P<0.05); TMRE fluorescence intensity in TPEN+astragaloside IV+2-DG group was significantly decreased compared with astragaloside IV+2-DG group (P<0.05); TMRE fluorescence intensity in TPEN+2-DG group was significantly decreased compared with the control group ( P<0.05). Conclusion:Astragaloside IV can exert neuroprotective effects through Zn 2+ inhibiting endoplasmic reticulum stress-induced apoptosis and preventing mPTP opening in PC12 cells, whose mechanism may be related to Fis1 and Bap31 expressions.

Objective:To analyze the characteristics of the molecular transmission network of newly-diagnosed human immunodeficiency virus type 1 (HIV-1) infected individuals in Shaoxing City, Zhejiang Province, and to provide evidence for epidemic trend and prevention.Methods:The plasma samples from 423 antiretroviral-naive HIV-1/acquired immunodeficiency syndrome patients from August 2018 to December 2019 were collected, and the pol gene fragments of HIV-1 from 375 samples were amplified by reverse transcription polymerase chain reaction (PCR) and nested PCR. The phylogenetic tree was constructed to analyze the molecular transmission network for subtypes and different gene distances by MEGA 6.0 software, HyPhy software and Cytoscape 3.7.2. Mutations on drug resistance was analyzed by online software tool of the HIV drug resistance database of Stanford University. Results:Eight subtypes were found in the 375 samples. Circulating recombinant form (CRF)07_BC(215/375, 57.33%) and CRF01_AE(103/375, 27.47%) were the major subtypes, followed by CRF85_BC, CRF55_01B, B, C, and CRF01_AE/B subtypes. One hundred and ninety-four individuals (51.73%) were connected to the transmission network at 1.50% genetic distance with 24 clusters. One hundred and twenty-nine individuals (34.40%) were connected to the transmission network at 0.75% genetic distance with 30 clusters, and 35 elderly patients were clustered in CL1.Forty-two cases had surveillance drug resistance mutation (SDRM), the prevalence of transmitted drug resistance was 11.20%(42/375). Thirty-eight cases had the drug mutations to non-nucleoside reverse transcriptase inhibitor, including K103 N(32/375, 8.53%), K103 S(4/375, 1.07%), Y188 L(1/375, 0.27%) and G190 A(1/375, 0.27%); four cases had the mutations to protease inhibitor, including M46 I(3/375, 0.80%) and V82 A(1/375, 0.27%). The cluster C2 sequences carried a high proportion of resistant mutations (94.29%, 33/35). Conclusions:HIV-1 subtypes in Shaoxing City are diverse and the CRF07_BC subtype spreads rapidly. The elderly patients with drug resistance genes in cluster CL1 at 0.75% gene distance need to be intervened immediately to prevent the drug resistance virus spread.

Objective: Buyinqianzheng Formula (BYQZF) is clinically employed in traditional Chinese medicine to treat Parkinson's disease (PD) by improving mitochondrial dysfunction. However, the underlying mechanisms by which BYQZF affects mitochondrial morphology remain unknown. Therefore, we observed the effects of BYQZF on mitochondria from the perspective of the PINK1/Parkin pathway. Methods: Cell survival rates were assessed by Cell Counting Kit-8 assay. Expression levels of PINK1 and Parkin mRNA were examined by qRT-PCR. Protein expression levels of PINK1, PINK1-Ser228, Parkin, Parkin-Ser65, Drp1, and Drp1-Ser637 were examined by western blotting. PINK1, Parkin, and Mito-Tracker? Red CMXRos (MTR) were stained by triple-labeled immunofluorescence, and observed under laser confocal microscopy. Results: Cell survival rate, mitochondrial form factor, mean length and number of mitochondrial network branches, mitochondrial activity, mRNA expression levels of PINK1 and Parkin, and protein expression levels of PINK1, Parkin, and Drp1-Ser637 were reduced after 1-methyl-4-phenylpyridinium (MPP+) intervention. In contrast, Pearson's correlation coefficients between PINK1 and Parkin, and between Parkin and MTR, as well as protein expression levels of PINK1-Ser228, Parkin-Ser65, and Drp1 increased significantly after MPP+intervention. Treatment with BYQZF increased cell survival rate, mitochondrial form factor, mean length and number of mitochondrial network branches, mitochondrial activity, mRNA expression levels of PINK1 and Parkin, and expression of PINK1, Parkin, and Drp1-Ser637 proteins. Pearson's correlation coefficients between PINK1 and Parkin, and between Parkin and MTR, as well as protein expression levels of PINK1-Ser228, Parkin-Ser65, and Drp1 decreased after BYQZF treatment. Conclusion: These results demonstrate that BYQZF has a protective effect on mitochondrial molecular mechanisms in the PD cell model, and the mechanism is related to the PINK1/Parkin pathway.

BACKGROUND: We investigated the effect of vitamin D deficiency on cardiovascular risk profiles in an Asian population with chronic kidney disease (CKD). METHODS: A total of 210 participants (62 non-dialysis CKD patients and 148 hemodialysis [HD] patients) were enrolled between December 2009 and February 2010. Vitamin D deficiency was determined using the serum 25-hydroxyvitamin D [25(OH)D] concentration. Blood pressure and arterial stiffness were measured. Subjects were divided into groups according to 25(OH)D concentration based on a cut-off of 13.5 ng/mL in non-dialysis CKD patients and 11.3 ng/mL in HD patients. RESULTS: The mean age was 61.7±12.3 years in non-dialysis CKD patients and 57.0±12.7 years in HD patients. In the non-dialysis CKD group, mean estimated glomerular filtration rate (eGFR) was 29.7±15.4 mL/min/1.73 m2. Mean 25(OH)D concentration was 13.6±7.8 ng/mL in non-dialysis CKD patients and 11.3±6.7 ng/mL in HD patients. More than half of the subjects had vitamin D deficiency (67.6% in non-dialysis CKD patients and 80.4% in HD patients). There were no significant differences in systolic blood pressure, pulse pressure, and arterial stiffness between higher and lower 25(OH)D groups among non-dialysis CKD and HD patients. Multivariate analysis revealed that female sex (odds ratio [OR]: 5.890; 95% confidence interval [CI]: 2.597–13.387; p<0.001) and presence of diabetes (OR: 2.434; 95% CI: 1.103–5.370; p=0.028) were significantly associated with lower serum 25(OH)D levels in HD patients. CONCLUSION: The prevalence of vitamin D deficiency was high in both nondialysis CKD patients and HD patients. Serum 25(OH)D concentration was not a significant factor associated with blood pressure and arterial stiffness among non-dialysis CKD and HD patients.
Asian Continental Ancestry Group ; Blood Pressure* ; Female ; Glomerular Filtration Rate ; Humans ; Multivariate Analysis ; Prevalence ; Renal Dialysis ; Renal Insufficiency, Chronic* ; Vascular Stiffness* ; Vitamin D Deficiency

BACKGROUND: Thoracolumbar segments (T11-L2) prone to damage due to its special anatomical and biomechanical characteristics. Therefore, fully understanding the shape of pedicle of vertebral arch and finding the visible, constant, and the point of insertion of the pedicle axis is very important to the safety of pedicle screw placement.OBJECTIVE: To measure the surgically relevant parameters of thoracolumbar pedicles between Han and Uygur males using computed tomography (CT) scan to provide some anatomic reference data for pedicle screw fixation.METHODS: The vertebral bodies and pedicles of adult males were scanned (60 cases of Han people and 60 cases of Uygur people) from T10-L3 with CT. The parameters were processed by three-dimensional reconstruction. Transverse pedicle width, pedicle axis length, transverse pedicle angle, and sagittal pedicle angle were measured by using length and angle measurement tool of browser in bone-window CT images. The age and stature information were recorded. All the data above were processed by SPSS 13.0 software.RESULTS AND CONCLUSION: (1) The mean transverse pedicle width of T12 and L1 in Han was bigger than that in the Uygur. (2) The mean transverse pedicle angle of T12 and L2 in Han was bigger than that in the Uygur. (3) The mean pedicle axis length and sagittal pedicle angle of T11 and L1 in Han were bigger than that in the Uygur (P < 0.05). (4) There were some differences among some parameters of the thoracolumbar pedicles between the Han and Uygur people.Data from any study only can be used as a guide for pedicle screw fixation. Preoperative CT evaluation may provide an individualized strategy to reduce the incidence of postoperative complications caused by misplacement.

OBJECTIVETo investigate the effects of the furin inhibitor α1-PDX on the growth, invasion, and tumorigenicity of cervical cancer cells and explore the mechanisms.

METHODSThe changes in the growth, migration and invasion of α1-PDX-transfected HeLa cells were observed using MTT assay, Boyden migration and invasion assay. The protein levels of furin and MT1-MMP were measured using Western blotting and furin activity was detected by enzyme activity assay in the transfected cells. HeLa cells were seeded subcutaneously in nude mice and the tumor volume changes were recorded.

RESULTSCompared with the control cells, α1-PDX-treated cells showed a significant growth inhibition by 18.4% at 24 h (P<0.01) with obviously lowered migration ability and cell invasiveness (P<0.01). Treatment with α1-PDX significantly reduced furin enzyme activity and MTI-MMP protein levels in HeLa cells. In nude mice, α1-PDX-treated HeLa cells exhibited a delayed and lowered tumorigenicity with reduced size of the tumors.

CONCLUSIONα1-PDX can inhibit the growth, metastasis and tumorigenicity of HeLa cells, the mechanism of which may involve a decreased furin activity and MTI-MMP expression.

Animals ; Female ; Furin ; antagonists & inhibitors ; HeLa Cells ; drug effects ; Humans ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Transfection ; Uterine Cervical Neoplasms ; pathology ; alpha 1-Antitrypsin ; pharmacology

BACKGROUND:Tissue inhibitor of matrix metaloproteinase 1 (TIMP-1) is the corresponding antagonist of matrix metaloproteinase 13 (MMP-13), and their balance between expression and functional activity exerts an important role in the metabolic state of the extracelular matrix. During the development of osteoarthritis, however, TIMP-1 and MMP-13 expressions and their expression ratio show unclear changes in DH guinea pigs.
OBJECTIVE:To explore the expression levels of MMP-13 and TIMP-1 in DH guinea pigs with different ages, and to analyze the relationship between the ratio of MMP-13 to TIMP-1 and the age-dependent degenerative changes in the articular cartilage.
METHODS:Twenty-four female Dunkin Hartley guinea pigs were sacrificed at age of 2, 4, 8, 12 months separately, with six animals at each time point. The knee joints were colected and gross visual appearance of the articular cartilage was observed, then were decalcified and prepared for paraffin sections. VG staining and Mankin score were used to analyze the histological changes. Immunohistochemistry was conducted to assess the expression levels of MMP-13 and TIMP-1 in the cartilage. Integrated absorbance values were used as the quantitive analysis calculated by Image pro-Plus 6.0. Linear regression analysis was done to analysis the relationship between Mankin score and the ratio of MMP-13/TIMP-1. RESULTS AND CONCLUSION:Normal appearance in the articular cartilage was observed in 2-month-old DH guinea pigs, while degenerative changes in the articular cartilage were shown in 4-month-old animals, which became severer with age. Significant difference was found in Mankin score between any two groups (P < 0.05). The ratio of MMP-13 to TIMP-1 increased with age, and the ratio was positively correlated to the Mankin score (P < 0.05). Age-related articular cartilage degeneration occurred in Dunkin Hartley guinea pigs at 4 months of age, and devoloped with age, which is related with the imbanlance of the expression ratio of MMP-13 to TIMP-1.

Objective To explore the value of endoscopic retrograde cholangiopancreatography (ERCP)in the treatment of chronic pancre-atitis (CP).Methods Thirty patients with CP who underwent ERCP from January 2008 to December 2012 were retrospectively analyzed. Serum amylase levels were determined before and at 24 hours after surgery.The reduction in abdominal pain was dynamically evaluated be-fore and after treatment.A follow -up of patients with steatorrhea and anxiety was performed.Regular reexamination using abdominal ima-ging or ERCP was performed to identify pancreatic lesions and conditions of pancreatic duct stents.Results All patients were successfully treated by ERCP and had pancreatic duct stents placed.The remission rate of abdominal pain reached 83.3% at 72 hours after surgery.One patient was assigned to surgery due to insignificant improvement in symptoms,and the case -fatality rate after surgery was 0.Conclusion ERCP is an effective method in the treatment of CP,which has the advantages of being safe,effective,and minimally invasive.

Objective To investigate the effects of the furin inhibitor α1-PDX on the growth, invasion, and tumorigenicity of cervical cancer cells and explore the mechanisms. Methods The changes in the growth, migration and invasion of α1-PDX-transfected HeLa cells were observed using MTT assay, Boyden migration and invasion assay. The protein levels of furin and MT1-MMP were measured using Western blotting and furin activity was detected by enzyme activity assay in the transfected cells. HeLa cells were seeded subcutaneously in nude mice and the tumor volume changes were recorded. Results Compared with the control cells, α1-PDX-treated cells showed a significant growth inhibition by 18.4% at 24 h (P<0.01) with obviously lowered migration ability and cell invasiveness (P<0.01). Treatment withα1-PDX significantly reduced furin enzyme activity and MTI-MMP protein levels in HeLa cells. In nude mice, α1-PDX-treated HeLa cells exhibited a delayed and lowered tumorigenicity with reduced size of the tumors. Conclusionα1-PDX can inhibit the growth, metastasis and tumorigenicity of HeLa cells, the mechanism of which may involve a decreased furin activity and MTI-MMP expression.

Objective To investigate the effects of the furin inhibitor α1-PDX on the growth, invasion, and tumorigenicity of cervical cancer cells and explore the mechanisms. Methods The changes in the growth, migration and invasion of α1-PDX-transfected HeLa cells were observed using MTT assay, Boyden migration and invasion assay. The protein levels of furin and MT1-MMP were measured using Western blotting and furin activity was detected by enzyme activity assay in the transfected cells. HeLa cells were seeded subcutaneously in nude mice and the tumor volume changes were recorded. Results Compared with the control cells, α1-PDX-treated cells showed a significant growth inhibition by 18.4% at 24 h (P<0.01) with obviously lowered migration ability and cell invasiveness (P<0.01). Treatment withα1-PDX significantly reduced furin enzyme activity and MTI-MMP protein levels in HeLa cells. In nude mice, α1-PDX-treated HeLa cells exhibited a delayed and lowered tumorigenicity with reduced size of the tumors. Conclusionα1-PDX can inhibit the growth, metastasis and tumorigenicity of HeLa cells, the mechanism of which may involve a decreased furin activity and MTI-MMP expression.