OBJECTIVE:To determine the plasma mycophenolic acid concentration by HPLC, and study the multidoses pharmacokinetics character of mycophenolic acid in Chinese kidney transplantation patients. METHODS: The samples were precipitated with acetonitrile before injection. Diamonsil C18 column was used. The mobile phase consisted of acetonitrile-10 mmol?L-1 KH2PO4 (5∶6) at a flow rate of 1.1 mL?min-1. The detection wavelength was 254 nm and the column temperature was 40 ℃. This method was used to determine the multidoses pharmacokinetics in 12 kidney transplantation patients. RESULTS: MPA was well-separated from internal standard in chromatography, and endogenous foreign substance in plasma had no interference on the determination. The liner range for MPA was 0.38～59.00 ?g?mL-1,and the lowest detectable concentration of MPA was 0.38 ?g?mL-1. The recovery rate stood at 89.32%～97.63%; Both intra-day and inter-day RSD were less than 8.00%. Significant individual difference was noted among the patients treated with MMF in pharmacokinetic results, which was in line with the literature. CONCLUSION: This method is accurate and simple and applicable for the pharmacokinetics study of mycophenolic acid.

The aim of this research is to investigate the influence of microencapsulation on the expression of the oxidative stress genes and exogenous regulation of HepG2 cells. We compared the expression of hemeoxygenase-1 (HO-1) and glutathione S-transferases-A1 (GST-A1) in HepG2 cells under different culture conditions through real-time PCR. The effects of exogenous antioxidants on cell viability and albumin levels were also evaluated through MTT assay and ELISA assay. The results showed that after culturing for 6 and 16 days, the expression levels of HO-1 in encapsulated cells were approximately 4.9 and 3.1 times higher than that of monolayer cells at the same culture period; As for the expression levels of GST-A1, they were elevated to 11.2 and 33 times of monolayer cells (P < 0.05). Accordingly, we found that NAC at 5-10 mmol/L significantly increased the viability by 40%-70% and the biosynthetic function by 20%-30% in microencapsulated HepG2 cells (P < 0.05). GSH increased the viability of the encapsulated cells by 20%-55% and the biosynthetic function by 15% (P < 0.05). In conclusion, oxidative stress exists in the microcapsules and affects genes expression. Exogenous antioxidants can prevent the inhibition effects of oxidative stress on cellular growth.
Antioxidants ; pharmacology ; Cell Survival ; Gene Expression Regulation ; Glutathione Transferase ; metabolism ; Heme Oxygenase-1 ; metabolism ; Hep G2 Cells ; Humans ; Oxidative Stress

OBJECTIVETo investigate the effects of small interfering RNA (siRNA) targeting YAP on the proliferation and apoptosis of human periodontal ligament stem cells (hPDLSCs).

METHODSSynthesized sequences of siRNA were transfected into hPDLSCs by Lipofectamine™ 2000. The expression of YAP was identified by using real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. Proliferation activity was detected by using cell counting kit-8 (CCK-8). Changes in the cell cycle and apoptosis rate were detected by using flow cytometry. Results were analyzed by using SPSS 19.0, and P < 0.05 was considered statistically significant.

RESULTSExpression of YAP mRNA and protein were significantly downregulated after 48 h of transfection (P < 0.001). No obvious difference was found in the expression levels of YAP protein between 48 and 72 h, thus indicating that siRNA could inhibit the expression of YAP persistently and effectively. Proliferation activity was inhibited, and apoptosis rate was increased. Cell cycle was changed as the proportion of G₁and S phases increased (P < 0.01) and G₂ phase decreased (P < 0.05).

CONCLUSIONKnocking down YAP gene by siRNA could inhibit proliferation activity, induce apoptosis, and change the cell cycle of hPDLSCs. Thus, YAP could regulate the proliferation and apoptosis of hPDLSCs.

Adaptor Proteins, Signal Transducing ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Down-Regulation ; Humans ; Periodontal Ligament ; drug effects ; Phosphoproteins ; RNA, Messenger ; RNA, Small Interfering ; pharmacology ; Stem Cells ; drug effects ; Transfection

ObjectiveTo study the role of Kukoamine B (KB) in inhibiting the inflammatory response of small intestine in septic mice and its molecular mechanisms.Methods Twenty-four male ICR mice were randomly divided into control group, model group, and KB intervention group (each,n= 8). Sepsis model was reproduced by intra-peritoneal injection of 20 mg/kg lipopolysaccharide (LPS), while equivalent normal saline was given in control group, and 20μg/kg KB was injected through caudal vein 4 hours after LPS challenge in KB intervention group. The blood/tissue samples (jejunum and ileum) were harvested 8 hours after LPS injection. The levels of plasma LPS, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were measured. The pathological changes in small intestine tissues were observed under light microscope, while the levels of inflammatory cytokines TNF-α and IL-1β in the tissue homogenates (jejunum and ileum) were assessed by enzyme linked immunosorbent assay (ELISA). The activity of myeloperoxidase (MPO) was measured by colorimetry. The expression of intercellular adhesion molecule-1 (ICAM-1) was determined by immunohistochemistry. The expressions of inducible nitric oxide synthase (iNOS) mRNA was assayed by reverse transcription-polymerase chain reaction (RT-PCR). The activation of nuclear factor-κΒ (NF-κΒ) was determined by Western Blot.Results The mice in model group were found to have an increase in microvascular permeability, interstitial edema, and infiltration of white blood cells, and the levels of LPS, TNF-α and IL-1β in their plasma, with an increase in concentrations of TNF-α and IL-1β, activity of MPO, positive expression of ICAM-1, expression of iNOS mRNA and NF-κB protein in small intestine (jejunum and ileum). Compared with model group, in mice with KB intervention, microvascular permeability, interstitial edema, and infiltration of white blood cells were reduced significantly, while the levels of LPS, TNF-α and IL-1β in plasma, the concentration of TNF-α and IL-1β, the activity of MPO, the positive expression of ICAM-1, the expression of iNOS mRNA and NF-κB protein in small intestine (jejunum and ileum) were significantly decreased [plasma LPS (kEU/L): 654.09±28.13 vs. 1 155.65±47.15, TNF-α (ng/L): 12.75±0.47 vs. 30.61±0.71, IL-1β (ng/L): 53.06±5.32 vs. 64.47±2.61; jejunum TNF-α(ng/L): 43.27±1.20 vs. 64.82±2.09, IL-1β (ng/L): 326.38±14.47 vs. 535.22±13.48, MPO (U/g): 0.14±0.01 vs. 0.32±0.02, iNOS mRNA (2-ΔΔCt): 2.39±0.13 vs. 10.80±0.22, NF-κB protein (gray value): 0.687±0.062 vs. 1.404±0.046; ileum TNF-α (ng/L): 62.75±3.92 vs. 104.24±2.82, IL-1β(ng/L): 408.06±1.70 vs. 521.97±1.16, MPO (U/g): 0.36±0.08 vs. 0.66±0.05, iNOS mRNA (2-ΔΔCt): 1.65±0.11 vs. 3.59±0.29, NF-κB protein (gray value):0.830±0.114 vs. 1.609±0.051, allP< 0.05].Conclusion KB can combine with LPS and inhibit LPS/Toll-like receptor 4 (TLR4) signaling pathway, thereby significantly inhibit the inflammatory response and protect the function of the small intestine in LPS-induced septic mice.

Objective To observe the clinical efficacy and toxicity of intraperitoneal chemotherapy combined with intravenous chemotherapy in treatment of cervical cancer.Methods The experimental group of 40 patients received systemic chemotherapy of docetaxel and cisplatin combined abdominal intraperitoneal high-frequency hyperthermia in the control group,40 patients received docetaxel plus cisplatin chemotherapy,The efficacy and toxicity were observed.Results The complete remission rate of observotion group was 82.5％,significantly higher than 62.5％ in control group.Effective rate in observation group was 92.5％ and also significantly higher than 75.0％ in control gronp.After 6 cycles of chemotherapy,the improvement of the experimental group was 85.0％ higher than 57.5 ％ in control gronp.The differemles between the two groups was statistically significant(P ＜ 0.05); the main side effects during chemotherapy are gastrointestinal reactions,since the cells decreased,hair loss,the difference was not statistically significant(P ＞0.05).Conclusion The chemotherapy of docetaxel and cisplatin intraperitoneal chemotherapy can significantly improve the treatment of cervical cancer,without increasing side effects.

Objective To study the application of 99Tcm in rabbit cerebral thromboembolic stroke and thrombolysis effect of recombinant tissue plasminogen activator (rt-PA).Methods The 0.5 mL radioactive pertechnetate sodium (specification:5 mCi/2mL and radiation intensity 92.5 MBq/mL) was combined with 30 μL stannous chloride (5 mg/mL),and the 20 μL mixture was joined to whole blood,red blood cells,and plasma for labelling.Then 50 μL CaCl2 (0.5 mol/L) and bovine thrombin (50 IU/mL) were doped in mixture,and rapidly sucked into a polyethylene plastic pipe (PE80).Thrombus was formed for 2 h at 37 ℃ and cut into small pieces of 10 mm.Autologous blood clots combined with 99Tcm from external carotid artery were injected to internal carotid artery of rabbit,the radioactivity (counts per minute,CPM) was measured by gamma counting instrument,and the improvement of rt-PA 4.5 mg/kg (clinical equivalent dose) on this model was observed.Results After thromboembolism,CPM increased approximately by (5.1 ± 1.3) times,which suggested that the model was reliable.The rt-PA 4.5 mg/kg had significant progressive thrombolysis effect.Conclusion 99Tcm tracer technology could be applied to rabbit cerebral stroke model,which is stable and reliable

Objective To investigate the inhibitory effect of kukoamine B (KB) on lung inflammatory responses in mice with sepsis and its possible molecular mechanism.Methods Twenty-eight male mice were randomly divided into control group (n=8),lipopolysaccharide (LPS) group (n=10),and LPS + KB group (n=10).Sepsis model was reproduced by intra-peritoneal injection of 20 mg/kg LPS,while equivalent normal saline was given in control group,and 20 μg/kg KB was injected through caudal vein 4 hours after LPS challenge in LPS + KB group.After 8 hours of LPS challenge,the concentration of LPS in plasma and the activity of myeloperoxidase (MPO) in the lung tissue were determined.The contents of tumor necrosis factor-α (TNF-α) and interleukin-lβ (IL-1β) in plasma,alveolar lavage fluid and lung tissue homogenates were assessed by enzyme linked immunosorbent assay (ELISA).The activation of nuclear factor-κB (NF-κB) and the expression of inducible nitric oxide synthase (iNOS) in lung tissue were determined by Western Blot.The pathological changes in lung tissues were observed with hematoxylin-eosin (HE) staining.The expression of intercellular adhesion molecule-1 (ICAM-1) in lung tissue was determined by immunohistochemistry.Results Compared with control group,the concentration of LPS in plasma (kEU/L:1 155.650 ± 147.149 vs.31.390 ± 18.859),MPO activity (U/g:1.177 ±0.093 vs.0.775 ±0.166),NF-κB activity (gray value:1.557 ±0.105 vs.0.824 ±0.032) and the expression of iNOS (gray value:0.650 ±0.129 vs.0.392 ±0.097) were significantly increased in LPS group (all P＜0.05).After KB intervention,the concentration of LPS (624.461 ± 149.012),MPO activity (0.919 ±0.023),NF-κB activity (1.127 ±0.074) and the expression ofiNOS (0.425 ± 0.066) were significantly lowered (all P＜0.05).Compared with control group,the contents of TNF-α (ng/L:47.325 ± 13.864 vs.6.534 ± 0.544,13.382 ± 2.231 vs.3.748 ± 0.692,31.127 ± 7.399 vs.14.948 ± 4.673) and IL-1β (ng/L:74.329 ± 11.890 vs.29.921 ± 6.487,9.422 ± 2.674 vs.1.105 ± 0.364,528.509 ± 32.073 vs.109.945 ± 13.561) in plasma,alveolar lavage fluid and lung tissue homogenates were obviously enhanced in LPS group (all P＜0.05).With KB intervention,the contents of TNF-α (20.331 ± 7.789,7.145 ± 1.202,15.966 ± 2.946) and IL-1β (57.707 ±8.098,2.212 ± 0.878,426.154 ± 11.270) were markedly reduced (plasma TNF-α:F=16.052,P=0.002; IL-1β:F=20.649,P=0.000; lung tissue homogenates TNF-α:F=31.134,P=0.001; IL-1β:F=22.792,P=0.002;alveolar lavage fluid TNF-α:F=10.013,P=0.009; IL-1β:F=319.857,P=0.000).In addition,leukocyte infiltration to the lung tissue was attenuated,and the expression of ICAM-1 was reduced by KB in histological examination.Conclusion KB,as a neutralizer of LPS,can inhibit the release of inflammatory mediators,reduce the pulmonary inflammatory response and protect the function of lung in septic mice.

Objective:To compare the effects of different anesthesia methods on postoperative analgesia, short-term cognitive function and deep venous thrombosis(DVT) in patients undergoing hip replacement.Methods:From March 2017 to March 2019, 96 patients with hip replacement were divided into group A (48 cases) and group B (48 cases) according to the random digital table method.Epidural anesthesia was used in group A, and general anesthesia was used in group B. The perioperative conditions, anesthesia effect, visual analogue scale (VAS) at postoperative 3, 12 and 24 hours, postoperative cognitive dysfunction (POCD) and DVT at postoperative 1, 3 and 7 days were compared between the two groups.Results:The operation time, intraoperative bleeding volume and time to get out of bed between the two groups had no statistically significant differences (all P>0.05). The recovery time[(14.98±2.31)min]and extubation time[(17.35±3.54) min]of group A were faster than those of group B[(19.13±3.42)min and (22.18±2.87)min]( t=6.967, 7.343, all P<0.05). The VAS scores of group B at postoperative 3 h[(3.27±0.73)points], postoperative 12 h[(2.45±0.38)points]and postoperative 24 h[(1.98±0.45)points]were higher than those of group A[(2.34±0.56)points, (1.65±0.43)points and (1.07±0.31)points]( t=7.003, 9.659, 11.538, all P<0.05). The incidences of POCD in group A at postoperative 3d(10.42%) and 7d(2.08%) were lower than those in group B (33.33% and 14.58%) (χ 2=7.357, 3.909, all P<0.05). The incidences of DVT in group A at postoperative 3d(4.17%) and 7d(8.33%) were lower than those in group B (20.83% and 29.17%) (χ 2=7.357, 3.909, all P<0.05). Conclusion:Epidural anesthesia is superior to general anesthesia in hip replacement.Postoperative analgesia is superior to general anesthesia, and can reduce the occurrence of short-term POCD and DVT, which is worthy of clinical reference.

Objective:To amendment a tool of theory of mind (TOM) tests that can be applied to children with autism spectrum disorder (ASD) in mainland China,and assess the ability of ToM of children with ASD.Methods:The items of the ToM tests were revised by observing and recording events of children's life.Totally 200 normal children were selected in line with the standard from kindergarten to Grade 6 for formal tes-ting.With 156 valid data,Pearson correlation coefficient and Cronbach α coefficient were established for the test.Three experts were invited to make sure the content validity.Researchers randomly selected 30 normal children for retest purpose after 2 months.Twenty five children with ASD were tested and compared with normal children's test scores.Results:The revised test included 39 entries,which was divided into three sub-tests,42 points in total.The correlation coefficient of three subtests of the tests was 0.54-0.77,the correlation coefficient between the test and the subtest was 0.62-0.93 (Ps ＜ 0.01).The scores of three experts for the test were 114,108,and 105.The total scores and subtest scores were lower in children with ASD than in normal children (Ps ＜0.01).The Cronbach a coefficient of the test was 0.84,Cronbach a coefficients for three subtest were 0.83,0.80,and 0.78,respectively.The retest reliability was 0.84,and reliabilities for three subtest were 0.75,0.74,and 1.00.Conclusion:The revised Theory of Mind Tests for Children with Autism Spectrum Disorder are fulfilled mostly in line with psychometric testing requirement.It might be a selection to measure the ability of theory of mind of children with ASD in mainland China.

Objective:To construct an early comfort assessment and management plan for ICU patients with high flow nasal cannula, and to provide reference for clinical nursing practice and management.Methods:The domestic and foreign databases were retrieved, and the literatures on high flow nasal cannula were obtained and evaluated. The first draft of the early evaluation and management scheme of nasal high-flow humidification oxygen therapy was developed. Two rounds of expert consultation were conducted by Delphi method.Results:The early comfort assessment and management plan for ICU patients with high flow nasal cannula was formed, including 3 primary indicators, 12 secondary indicators, and 32 tertiary indicators. The authoritative coefficients of 2 rounds of expert consultation were 0.915 and 0.931, respectively. The coordination coefficients were 0.221, 0.130 and 0.278( χ 2 values were 8.000, 27.140, 163.824, P<0.05 or 0.01). Conclusions:The early comfort assessment and management plan of ICU patients with high flow nasal cannula are consistent with the advice of experts at all levels. The enthusiasm and authority of the experts is high, which provides a basis for the scientific management of ICU patients with nasal high-flow humidification oxygen therapy.