Objective To explore the effects of baicalin in the treatment of a preeclampsia (PE) rat model by detecting the expression of X-linked inhibitor of apoptosis protein (XIAP) and cysteine containing aspartate-9 (Caspase-9) and observing the ultrastructure of mitochondria in trophoblast cells.Methods Forty-eight pregnant Wistar rats were randomly divided into two groups:12 in the control group and 36 in the PE model group.The PE model was established with subcutaneous injection of l-nitro arginine methyl ester with 100 mg/kg per day from the 13th day of pregnancy.Beginning from the 16th day of pregnancy,the PE rats were injected with different doses of baicalin till cesarean section,and divided into three groups:non-intervention PE model group treated with saline (NIP group),low-dose baicalin intervention group (IDB group) at 50 mg/kg per day,and high-dose baicalin intervention group (HDB group) at 100 mg/kg per day.The rat tail artery blood pressure and 24-h urine protein level were measured at pregnant day 10,16 and 20.The levels of XIAP and Caspase-9 in placenta were measured by immunohistochemistry.The ultrastructure of mitochondria of trophoblast cells of the rat placenta was observed under electron microscope.T test,F test and LSD-t test were applied for statistical analysis.Results (1) On pregnant day 10,no significant differences were observed in rat tail artery systolic blood pressure,diastolic blood pressure and 24-h urine protein level between the control group and PE model group (all P＞0.05).On pregnant day 16 and 20,the systolic blood pressure,diastolic blood pressure and 24-h urine protein level of NIP,IDB and HDB groups were significantly higher than those of control group [pregnant day 16:systolic blood pressure:(137.74±5.21),(136.15±4.86),(138.28±4.79) and (110.57±3.79) mmHg (1 mmHg=0.133 kPa),diastolic blood pressure:(89.58 ± 5.50),(88.45 ± 8.59),(89.42 ± 6.29) and (80.28 ± 7.36) mmHg,24-h urine protein:(7.78 ± 0.45),(7.53 ± 0.54),(7.86± 0.57) and (6.45 ± 0.56) mg;pregnant day 20:systolic blood pressure:(145.26 ± 4.67),(131.28 ± 4.34),(130.93 ± 5.33) and (110.40 ± 6.92) mmHg,diastolic blood pressure:(89.87±6.55),(85.34±7.33),(84.64±7.36) and (80.19±7.34) mmHg,24-h urine protein:(11.18±0.42),(9.65±0.54),(9.06±0.56) and (6.31 ±0.45) mg] (all P＜0.01).On pregnant day 20,the systolic and diastolic blood pressure and 24-h urine protein level in IDB and HDB groups were significantly lower than in NIP group (all P＜0.05),but showed no significant differences between IDB and HDB groups (allP＞0.05).(2) Compared with NIP group,the expression of XIAP in control group,IDB and HDB groups were significantly increased(210.39±0.78,180.56±0.82,195.36±0.96 and 192.84± 1.06,all P＜0.01).There was no significant difference in the expression of XIAP between IDB and HDB groups (P=0.66).The expression of Caspase-9 in control group,IDB and HDB groups were significantly decreased compared with NIP group (210.36±0.55,195.53±0.96,198.42± 1.01 and 185.25±0.64,all P＜0.01).There was no significant difference in the expression of Caspase-9 between IDB and HDB groups (P=0.65).Ultrastructure of mitochondria in NIP group showed different degrees of damage,matrix swelling,and mitochondrial cristae bresk or disappearance.In IDB group,mitochondrial matrix swelling was not obvious,and mitochondrial cristae were visible.In HDB group,mitochondrial cristae were neat and clear.Conclusions Baicalin may play an important role in the treatment of preeclampsia by reversing the trophoblast apoptosis and improving the ultrastructure of mitochondria through its regulation of XIAP expression and downregulation of Caspase-9 expression.

BACKGROUND:Human embryonic stem cels exhibit self-renewal and multi-differentiation potential, and can differentiate into endothelial cels under certaininduction conditions.
OBJECTIVE:To explore induced conditions of the human embryonic stem cels differentiating into endothelial cels and to investigate the effect of vascular endothelial growth factors on theendothelial differentiation of human embryonic stem cels.
METHODS:After resuscitation,passage40 human embryonic stem cel lines H9 weresubjected to suspension culture to prepare embryos, and after 5-day culture,these cels werecultured in attachment medium to differentiate into embryoid bodies,folowed by induction with50 μg/L vascular endothelial growth factors. Passage 2 and 15 embryonic stem cels after induced differentiation weretaken for Dil-Ac-LDL uptake test and immunohistochemical staining, respectively.
RESULTS AND CONCLUSION:After 1-day culture, cord-like or polygonal monolayer cels around embryoid bodies showed bud-like andradialgrowth witharelative rapid speed merging into surrounding colonies; at 2-3 days, the number of suspension cels increased further, but the smal-round cels in the center began to die; at 5 days, embryoid bodies started to passage, and aggregated cels exhibited typical paving stone-like appearance. Moreover, some human embryonic cels after induced differentiation could actively takeupfluorescent labeled LDL,andred fluorescent particlesappeared.Additionaly, passage 15 embryonic stem cels after induced differentiation could express CD31 and FLK-1.These findings suggest that human embryonic stem cels induced by vascular endothelial growth factors can differentiate into endothelial cels.

Objective To investigate the expression of matrixmetalloprotein -9(MMP-9)and E-cad-herin in triple negative breast cancer ( TNBC) and its relationship with the clinicopathologic features of TNBC .To analyze the function of MMP -9 and E-cadherin in TNBC .Methods Immunohistochemical method was used to detect the expression of MMP -9 and E-cadherin in 127 cases of TNBC.Results The MMP-9 positive rate was 53.54%and E-cadherin positive grade was 32.28%in TNBC.Expression of MMP-9 was correlated with tumor size(P=0.007),histological grade(P=0.006),TNM stage(P=0.003),lymph node metastasis(P=0.000)and lymph duct invasive(P=0.000).Expression of E-cadherin was correlated with lymph node metasta-sis(P=0.016)and lymph duct invasive(P=0.015).However,they were not correlated with other factors .Con-clusion The expression of MMP -9 and E -cadherin was correlated with invasion and metastasis in TNBC , which could be an important research topic in the future studies .

Objective This study analyzes the main technical fields and their mutual correlation concerning the international patents of acupuncture to provide decision support for the strategy of developing acupuncture related products.Methods Derwent Innovation Index was applied as the data source,Thomson Data Analyzer was adopted as the main analyzing tool,and the co-occurring clustering spectrum analysis method of international patent number (IPC) was used.Results Based on the retrieval and analyzed results,the main classification of international patent of acupuncture and the technical association diagram of Chinese medicine acupuncture patents were formed.Conclusion The main technical fields of acupuncture international patent included acupuncture needles,acupuncture instruments combined with drug and acupuncture instruments combined with modem physical methods as well as the point health care products.Among them,the methods applied in acupuncture,namely the magnetic therapy method,laser therapy,thermal therapy,roller and ball massage adopted in acupuncture point massage instruments which combine traditional herbal medicine,have already been the research hotspot of current acupuncture patent products.

Objective:To improve the standard and quality of clinical trials, the possible risks of Investigator-Initiated Clinical Trial(IIT) approvals based on drug supply and security were discussed and suggestions were put forward.Methods:According to the laws and regulations and literature review, concerning experimental drug supply and security during project negotiation, the risk points of IIT approvals were comprehensively analyzed and suggestions were put forward.Results:There are four main types of risks in assessing IIT approvals in terms of drug supply and security: drug entry and sales, drug promotion, discounts of observation fees, and concept confusion. Healthcare institutions should pay attention to and coordinate the IIT approvals.Conclusions:IIT is a supplement and extension of Industry Sponsored Trial(IST), which should be actively carried out by healthcare institutions while also paying attention to the security and risk prevention of drug supply, ensuring a standardized and orderly manner.

The activation and deactivation of Ca(2+)- and calmodulindependent neuronal nitric oxide synthase (nNOS) in the central nervous system must be tightly controlled to prevent excessive nitric oxide (NO) generation. Considering plasma membrane calcium ATPase (PMCA) is a key deactivator of nNOS, the present investigation aims to determine the key events involved in nNOS deactivation of by PMCA in living cells to maintain its cellular context. Using time-resolved Förster resonance energy transfer (FRET), we determined the occurrence of Ca(2+)-induced protein-protein interactions between plasma membrane calcium ATPase 4b (PMCA4b) and nNOS in living cells. PMCA activation significantly decreased the intracellular Ca(2+) concentrations ([Ca(2+)]i), which deactivates nNOS and slowdowns NO synthesis. Under the basal [Ca(2+)]i caused by PMCA activation, no protein-protein interactions were observed between PMCA4b and nNOS. Furthermore, both the PDZ domain of nNOS and the PDZ-binding motif of PMCA4b were essential for the protein-protein interaction. The involvement of lipid raft microdomains on the activity of PMCA4b and nNOS was also investigated. Unlike other PMCA isoforms, PMCA4 was relatively more concentrated in the raft fractions. Disruption of lipid rafts altered the intracellular localization of PMCA4b and affected the interaction between PMCA4b and nNOS, which suggest that the unique lipid raft distribution of PMCA4 may be responsible for its regulation of nNOS activity. In summary, lipid rafts may act as platforms for the PMCA4b regulation of nNOS activity and the transient tethering of nNOS to PMCA4b is responsible for rapid nNOS deactivation.
Animals ; Brain ; metabolism ; Calcium ; metabolism ; Cells, Cultured ; Cerebellum ; cytology ; Fluorescence Resonance Energy Transfer ; HEK293 Cells ; Humans ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type I ; metabolism ; PDZ Domains ; Plasma Membrane Calcium-Transporting ATPases ; metabolism ; Protein Interaction Maps ; Protein Isoforms ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective The potential mechanism of hepatotoxicity induced by rhubarb was preliminarily explored by network pharmacology and verified by cell experiments.Methods Based on network pharmacology,component collection and target prediction are carried out through multiple databases.PPI network construction,GO enrichment analysis and KEGG pathway analysis were combined with software to systematically predict the mechanism of hepatotoxicity induced by rhubarb.The pathway information predicted by network pharmacology was verified by primary hepatocyte experiments and Western blot experiments.Results The results of network pharmacology showed that RH was the main component of hepatotoxicity induced by rhubarb.Seventeen core targets of hepatotoxicity induced by rhubarb were obtained.KEGG results suggested that DNA damage and apoptosis were one of the key mechanisms of hepatotoxicity induced by rhubarb.The results of primary hepatocytes and Western blot showed that RH could inhibit the viability of primary hepatocytes in a time-dose dependent manner.ABT and SFP can significantly reduce the toxicity of RH on primary liver cells in mice,and RFP can increase the toxicity of RH to mouse primary liver cells.Upregulation of γ-H2AX and PARP-1 protein in primary liver cells of mice after treatment with different concentrations of RH.Conclusion RH in rhubarb can significantly inhibit the viability of mouse primary hepatocytes,and its toxicity to mouse primary hepatocytes is mainly caused by the metabolic activation of RH by CYP 2C9.RH can activate PARP-1 protein,phosphorylate H2AX,induce DNA damage and apoptosis in mouse primary hepatocytes.

Apoptosis ; Calcineurin ; metabolism ; Chymotrypsin ; metabolism ; Humans ; Lysosomes ; enzymology ; Mitochondria ; metabolism ; pathology ; Mitochondrial Dynamics ; Neuroblastoma ; metabolism ; pathology

This study was aimed to observe the curative effect and safety of Danzhi Jiangtang Capsule ( DJC ) combined with atorvastatin on carotid artery intima-media thickness (IMT) in diabetes patients without hyper-tension . A total of 196 diabetes patients without hypertension with incrassate carotid artery IMT were randomly divided into the control group ( 98 cases ) and the treatment group ( 98 cases ) . The conventional diabetes thera-py was given to both groups . The atorvastatin of 20 mg/night was given to the control group . And the atorvas-tatin 20 mg/night added with DJC 9 . 0 g/night were given to the treatment group . The treatment course was
12 months . Carotid artery IMT , carotid atherosclerotic plaque area , FPG , FIns , HOMA-IR , HbA1c , blood lipids , hepatorenal function and etc . were examined before and after the treatment respectively . The results showed that there was a significant positive correlation between carotid artery IMT and FIns , HOMA-IR , HbAlc , LDL-C . After 12-month treatment , the total effectiveness is 85 . 87% in the treatment group . And there was significant difference compared with the control group ( P < 0 . 05 ) . The levels of FPG , FIns , HOMA-IR , HbAlc of the treatment group had no difference compared with the control group . Compared with the control group, TC and LDL-C of the treatment group was obviously decreased (P < 0.05). And HDL-C was significantly increased ( P < 0 . 05 ) . The carotid artery IMT of the treatment group decreased from ( 0 . 11 ±0 . 01 ) cm to ( 0 . 08 ± 0 . 01 ) cm . And compared with the control group , there was statistical significance ( P <0 . 05 ) . The carotid atherosclerotic plaque area of 58 cases in the treatment group decreased from ( 0 . 37 ±0.56) cm2 to (0.21 ± 0.25) cm2. However, there was no statistical significance compared to the control group. There were 5 adverse events in the control group and 9 adverse events in the treatment group . And there was no difference between two groups. It was concluded that DJC combined with atorvastatin can regulate lipid metabolism and reduce carotid artery IMT .

Objective To observe the dynamic characteristics of hepatitis B core antibody (anti-HBc) titers in chronic hepatitis B (CHB) patients treated with interferon and to explore the predictive value of anti-HBc for response to interferon.Methods The clinical information of the patients diagnosed with CHB in Department of Infectious Diseases , the First Affiliated Hospital of Xi′an Jiaotong University from October 2011 to October 2014 were collected.HBV DNA, liver function and HBV serological markers of CHB patients were tested dynamically during and after interferon treatment.The dynamic characteristics of anti-HBc titers in patients with different virological responses were analyzed.The predictive values of anti-HBc titer for the efficacy of interferon treatment of CHB patients were analyzed by binary logistic regression .Results Of the 42 CHB patients aging(30.8 ±10.1) years old, 34 patients were hepatitis B e antigen (HBeAg) positive and 8 were negative.All patients completed 48-week interferon treatment and 24-week follow-up after the end of treatment. Among them, 28.6%( 12/42), 26.2%( 11/42 ) and 45.2%( 19/42 ) of patients achieved sustained virological response (SVR), virological relapse ( VR) and non-response ( NR), respectively.Patients with different virological response presented various characteristics of anti -HBc titers.Compared with NR group, the anti-HBc titers at baseline and week 12 were significantly higher in SVR group (at baseline: [4.93 ±0.30] vs [4.70 ±0.33] lg IU/mL, t ＝2.147, P ＝0.013; at week 12: [4.83 ± 0.23] vs [4.44 ± 0.41] lg IU/mL, t＝3.032, P＝0.007).The anti-HBc titers in SVR group at week 12 and week 24 were significantly higher than those in VR group (at week 12: [4.83 ±0.23] vs [4.67 ±0.51] lg IU/mL, t＝2.400, P＝0.039; at week 24: [4.73 ±0.21] vs [4.55 ±0.50] lg IU/mL, t＝2.542, P＝0.039).By multivariate logistic regression analysis, the anti-HBc titer at baseline was the independent predictive factor for SVR in CHB patients treated with interferon (OR＝6.000, 95%CI: 1.118 -20.486, P＝0.037).The area under receiver operating characteristics curve was 0.753 and the optimal cutoff value of anti-HBc titer for the response to interferons in CHB patients was 5.03 lg IU/mL, with positive predictive value of 64.3%and negative predictive value of 89.3%.Conclusions Dynamic pattern of anti-HBc titers is correlated with different virological responses in CHB patients treated with interferon , and the baseline anti-HBc titer is the independent predictive factor for SVR.