Objective: To optimize the matrix prescription of Compound Qingfengteng Cataplasm (CQC). Methods: Taking the comprehensive senses as evaluation index, the types and dosage of the matrix were investigated by single factor. Further combined with the initial viscosity and holding viscosity index to optimize the matrix prescription of CQC by orthogonal test, and in vitro transdermal absorption experiments were carried out by Franz diffusion pool method to screen penetration enhancers. Results: The optimal formulation of CQC was as follows: 2.5 g of NP-700, 11 g of glycerin, 0.08 g of glycine aluminum, 0.08 g of tartaric acid, 0.8 g of PVP K90, 0.5 g of PVPP, 20 g of water, and 4.2 g of dry extract. 3% nitrile as a penetration agent could enhance penetration action. Conclusion: The preferred matrix prescription is practicable; CQC adhesion is moderate, no residue, paste traits, skin followability are good.

Objective: To determine the prevalence of human cytomegalovirus (HCMV) infection and plasma level of its neutralizing antibody in hypertension patients, so as to discuss the association between HCMV infection and hypertension. Methods: A total of 51 anti-coagulated blood samples were collected from hypertension patients and the 50 control samples were obtained from age- and sex-matched healthy controls in the Second People's Hospital of Anhui Province from Jan. 2013 to Apr. 2013. Neutralizing antibody level was analyzed by rapid fluorescence micro-neutralization; plasma anti-HCMV IgG and anti-HCMV IgM were determined by enzyme-linked immuno sorbent assay (ELISA), and HCMV UL93 DNA was detected by nested PCR. Results: The positive rates of HCMV UL93 DNA and anti-HCMV IgG in hypertension patients were significantly higher than those in healthy controls (72.55% vs 56.00% for UL93 DNA and 70.59% vs 50.00% for IgG, P<0.05). The positive rates of anti-HCMV IgM were not significantly different between the two groups (3.92% vs 2.00%, P>0.05). The geometric mean titer of HCMV neutralizing antibody was 38.56 ±20.42 in hypertension patients, which was significantly lower than that in healthy controls (60.12 ± 25.38, P = 0.0326). Conclusion: HCMV positive rate is higher in hypertension patients than in healthy controls; however, the specific neutralizing antibody level of HCMV is greatly lower than that in healthy controls, suggesting that the humoral immune status is declined in hypertension patients and there is a correlation between HCMV infection and hypertension.

Animals ; Diabetes Mellitus, Type 2 ; Diabetic Nephropathies ; Disease Models, Animal ; Humans ; Mice ; Rats

Objective To construct human embryonic stem cell (hESC) line with forkhead box G1 (FOXG1) gene knockout by clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing technology, and to investigate the role of FOXG1 gene in the early neural induction of hESCs. Methods Two guide RNAs (gRNAs) were transfected to induce FOXG1 gene large fragment knockout in hESCs by CRISPR/Cas9 gene editing technology. FOXG1 gene knockout hESCs were confirmed by monoclonal screening, sequencing and Western blotting analysis. The expression of the key markers including paired box 6 (PAX6), sex-determining region Y-box 2 (SOX2) and orthodenticle homeobox 2 (OTX2) was detected by immunofluorescence staining and qRT-PCR in the early process of neural induction before and after FOXG1 gene knockout. Results hESCs with FOXG1 gene large fragment knockout were successfully obtained by CRISPR/Cas9 gene editing technology. The results of immunofluorescence staining and qRTPCR suggested that FOXG1 deletion did not significantly influence the expression of PAX6, SOX2 and OTX2 during neural induction. Conclusion FOXG1 gene large fragment knockout in hESCs can be quickly induced by a pair of gRNAs cotransfection. FOXG1 deletion has no significant impacts on neural induction of hESCs.

OBJECTIVETo provide basis for making the quality standard of medicinal chrysanthemums [Dendranthema morifolium (Ramat.) Tzvel.].

METHODThe character of pharmacognosy.

RESULTThe characters of many of chrysanthemums as well as the differences between them have been clarified.

Chrysanthemum ; anatomy & histology ; classification ; Flowers ; anatomy & histology ; Pharmacognosy ; Plants, Medicinal ; anatomy & histology ; Quality Control ; Species Specificity

Objective: To observe the clinical efficacy of intraperitoneal injection of recombinant human vascular endostatin combined with 5-fluorouracil (5-FU) in patients with malignant ascites, and to detect the change of percentage of endothelial cells (ECs) in ascites before and after treatment, in order to explore the clinical value of endothelial cells in treatment of malignant ascites. Methods: Thirty-nine patients with malignant ascites received two cycles of intraperitoneal inject of endostar combined with 5-FU treatment. The clinical response, change of quality of life (QOL) and the adverse events were assessed. The flow cytometry was used to detect the level of endothelial cells in malignant ascites before and after treatment. Results: In 39 patients, the objective response rate (complete response and partial response) was 43.6% (1 7/39); the rate of QOL improvement was 53.8% (21/39). The rates of grades 3-4 adverse events were low, including leukopenia in 2 patients (5.1%, 2/39), thrombocytopenia in 1 patient (2.6%, 1/39), anemia in 2 patients (5.1%, 2/39), nausea in 4 patients (10.3%, 4/39), and diarrhea in 1 patient (2.6%, 1/39). In 16 patients achieveing clinical response, the percentage of endothelial cells in malignant ascites before treatment was significantly higher than that after treatment [(0.22±0.06)% vs (0.12±0.08) %, P = 0.005]. In 21 patients achieveing improvement in QOL, the percentage of endothelial cells in malignant ascites before treatment was also significantly higher than that after treatment [(0.1 9±0.08)% vs (0.1 3±0.08) %, P = 0.032]. The percentages of endothelial cells in malignant ascites in patients not achieveing clinical response and improvement in QOL after treatment had no significant changes (P = 0.11 4, P = 0.359). Conclusion: Intraperitoneal inject of endostar combined with 5-FU treatment can effectively control the malignant ascites and improve the QOL. The level of endothelial cells in malignant ascites may become a clinical marker to predict the response to the treatment of malignant ascites.

Objective: To investigate the changes of whole brain functional connections in cognitively impaired patients with nasopharyngeal carcinoma after radiotherapy. Methods: Totally 76 patients with pathologically proved nasopharyngeal carcinoma were collected, including 20 in radiotherapy-cognitive impairment group (RT-CI group), 34 in radiotherapy-no-cognitive impairment group (RT-NO-CI group) and 22 in the no-radiotherapy group (NO-RT group). Patients' gender, age, years of education, radiotherapy time and total measurement of radiotherapy were recorded, and the Beijing Montreal Cognitive Assessment (MoCA) scale was used for cognitive function scoring. Resting brain function MRI were collected, and brain functional connections were analyzed to obtain abnormal brain areas in RT-CI group and RT-NO-CI group. The general data and MoCA scores of 3 groups were compared, and the relationships between Z value of abnormal brain functional connectivity areas and MoCA score were analyzed. Results: There was no significant difference of age (F=2.78, P=0.30), years of education (F=3.95, P=0.37)nor gender (χ2=3.11, P=0.33) among 3 groups. MoCA score of RT-CI group was lower than that of RT-NO-CI group and NO-RT group (both P<0.01), and MoCA score of RT-NO-CI group was lower than that of NO-RT group (P<0.01). Compared with RT-NO-CI group, there was positive correlation between Z value of 1 pair of brain region and MoCA scale score in RT-CI group (P<0.01). Compared with NO-RT group, Z values of 8 pairs of brain areas in RT-CI group were correlated with MoCA scale scores (all P<0.01). Compared with NO-RT group, Z values of 5 pairs of brain areas in RT-NO-CI group were correlated with MoCA scale scores (all P<0.01). In RT-CI group, brain areas with abnormal functional connections mainly located in parahippocampal gyrus, parietal occipital cortex, prefrontal cortex and default mode network area. In RT-No-CI group, brain areas with abnormal functional connections mainly located in hippocampal gyrus, prefrontal lobe and parietal cortex. Conclusion: Early cognitive impair in patients with nasopharyngeal carcinoma after radiotherapy might manifest as abnormal brain functional connectivity in some specific brain regions.

OBJECTIVE: To investigate the effect of electroacupuncture (EA) on the p38 mitogen-activated protein kinase (p38 MAPK) and signal transducer and activator of transcription 3 (STAT3) pathway in hippocampus and frontal cortex of diabetic rats with cognitive impairment (CI), as well as the mechanism of EA in protection against CI in diabetic rats. METHODS: Thirty SD rats were divided into normal, model and EA groups (n=10 rats/group). The diabetic model was established by i.p.injection of Streptozotocin solution(25 mg/kg), followed by high-fat diet raising for 1 month, and the CI rats was confirmed by Morris water maze tasks. The rats in the EA group were given acupuncture at "Zusanli" (ST36) "Neiting" (ST44) and "Yishu" (EX-B3) 20 min/d, among which ST36 and ST44 were treated with EA. The treatment was conducted 6 times a week for 4 weeks. The fasting blood glucose (FBG) contents were assayed by glucometer before and after treatment. The rats' learning-memory ability was detected by Morris water maze tasks. The expression levels of IL-6、IL-1β、TNF-α、p38 MAPK、p-p38 MAPK、STAT3 and p-STAT3 in hippocampus and frontal cortex were detected by Western blot and quantitative real-time PCR, separately. The mean fluorescence intensity of p38 MAPK and STAT3 was observed by immunofluorescence histochemistry. RESULTS: After modeling, FBG and the escape latency of Morris water maze tasks were significantly increased in the model group compared with the normal group (P<0.001, P<0.01). Following EA treatment, the increased FBG and average escape latency were markedly reversed in the EA group relevant to the model group (P<0.05). Compared with the normal group, the proteins and mRNAs expression of IL-6, IL-1β, TNF-α, p38 MAPK, p-p38 MAPK, STAT3 and p-STAT3 in hippocampus and frontal cortex were significantly increased in the model group (P<0.001), as well as the mean fluorescence intensity of p38 MAPK and STAT3 in hippocampus and frontal cortex (P<0.001). Following EA intervention, the proteins and mRNAs expression of IL-6, IL-1β, TNF-α, p38 MAPK, p-p38 MAPK, STAT3 and p-STAT3, and the mean fluorescence intensity of p38 MAPK and STAT3 in hippocampus and frontal cortex were down-regulated(P<0.001, P<0.05). CONCLUSION: EA can inhibit the over production of pro-inflammatory cytokines in diabetic rats with CI, possibly by regulating the expression of p38 MAPK and STAT3 pathway.

Objective To observe the influences of regional citrate or systemic heparin anticoagulation on acid-base balance, coagulation, electrolytes, serum creatinine, alanine aminotransferase (ALT) and cardiac index (CI) during continuous renal replacement therapy (CRRT) rewarming in accidental hypothermia dogs. Methods Nineteen adult beagles were given abdominal trauma coupled with deep hypothermia [(28 0.5)°C] induced by seawater immersion for establishing animal model of trauma. According to rewarming ways, the animal models were randomly divided into three groups, comparison group (warm bath rewarming, n=5), CRRT rewarming-systemic heparin anticoagulation group (heparin anticoagulation group, n=7) and CRRT rewarming-regional citrate anticoagulation group (citrate anticoagulation group, n=7). During the rewarming routine blood examination was performed, and blood chemistry, coagulation function, blood gas and hemodynamic status were assayed, at the same time the mortality was recorded. Results During the rewarming, the mortality was 14.3% (1/7) in heparin anticoagulation group, 40.0% (2/5) in warm bath group and 0 in regional citrate anticoagulation group. Blood temperature increased to 38°C, the heparin anticoagulation group showed a significant decrease of platelet compared with citrate anticoagulation group and comparison group (P<0.05); The comparison group had a significant decrease of HCO3- compared with the other two groups (P<0.05). When the body temperature was increased to 34°C and 38°C, the comparison group showed significant increase of serum creatinine compared with the other two groups (P<0.05); the heparin anticoagulation group presented significant prolongation of APTT compared with citrate anticoagulation group and comparison group (P<0.05). When the body temperature increased to 30°C, 34°C and 38°C respectively, the citrate anticoagulation group had significantly increased serum calcium compared with heparin anticoagulation group and comparison group (P<0.05). There was no difference in serum sodium, ALT and CI between the three groups at each time point (P>0.05). Conclusions CRRT plus warm bath rewarming have better effect on improving metabolic acidosis than warm bath rewarming alone in accidental hypothermia dogs. In comparison with systemic heparin anticoagulation, regional citrate anticoagulation has smaller influences on coagulation system and platelet when CRRT rewarming performed in accidental hypothermia dogs.

BACKGROUNDOxidative Stress and p38 mitogen-activated protein kinase (p38MAPK) play a vital role in renal fibrosis. Pioglitazone can protect kidney but the underlying mechanisms are less clear. The purpose of this study was to investigate the effect of pioglitazone on oxidative stress and whether the severity of oxidative stress was associated with the phosphorylation level of p38MAPK.

METHODSRat mesangial cells were cultured and randomly assigned to control group, high glucose group and pioglitazone group. After 48-hour exposure, the supernatants and cells were collected. The protein levels of p22(phox), p47(phox), phosphorylated p38MAPK, total p38MAPK were measured by Western blotting. The gene expressions of p22(phox), p47(phox) were detected by RT-PCR. The levels of intracellular reactive oxygen species (ROS) were determined by flow cytometry. The levels of superoxide dismutase (SOD) and maleic dialdehyde (MDA) in the supernatant were determined respectively.

RESULTSCompared with the control group, the expression levels of p22(phox), p47(phox), phospho-p38 and ROS significantly increased, activity of SOD decreased in high glucose group, while the level of MDA greatly increased (P < 0.01). Pioglitazone significantly suppressed p22(phox), p47(phox) expressions and oxidative stress induced by high glucose. The expressions of p22(phox), p47(phox), phospho-p38MAPK and ROS generation were markedly reduced after pioglitazone treatment (P < 0.05). The activity of SOD in the the supernatant increased (P < 0.05), while the level of MDA decreased greatly by pioglitazone (P < 0.05). The level of oxidative stress was associated with the phosphorylation level of p38MAPK (P < 0.01).

CONCLUSIONPioglitazone can inhibit oxidative stress through suppressing NADPH oxidase expression and p38MAPK phosphorylation.

Animals ; Blotting, Western ; Glucose ; pharmacology ; Mesangial Cells ; drug effects ; enzymology ; NADPH Oxidases ; genetics ; metabolism ; Rats ; Reactive Oxygen Species ; metabolism ; Thiazolidinediones ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism