Molecular cloning and characterization of three phenylalanine ammonia-lyase genes from Schisandra chinensis / 中国天然药物

Phenylalanine ammonia-lyase (PAL), which catalyzes the conversion from L-phenylalanine to trans-cinnamic acid, is a well-known key enzyme and a connecting step between primary and secondary metabolisms in the phenylpropanoid biosynthetic pathway of plants and microbes. Schisandra chinensis, a woody vine plant belonging to the family of Magnoliaceae, is a rich source of dibenzocyclooctadiene lignans exhibiting potent activity. However, the functional role of PAL in the biosynthesis of lignan is relatively limited, compared with those in lignin and flavonoids biosynthesis. Therefore, it is essential to clone and characterize the PAL genes from this valuable medicinal plant. In this study, molecular cloning and characterization of three PAL genes (ScPAL1-3) from S. chinensis was carried out. ScPALs were cloned using RACE PCR. The sequence analysis of the three ScPALs was carried out to give basic characteristics followed by docking analysis. In order to determine their catalytic activity, recombinant protein was obtained by heterologous expression in pCold-TF vector in Escherichia coli (BL21-DE3), followed by Ni-affinity purification. The catalytic product of the purified recombinant proteins was verified using RP-HPLC through comparing with standard compounds. The optimal temperature, pH value and effects of different metal ions were determined. Vmax, Kcat and Km values were determined under the optimal conditions. The expression of three ScPALs in different tissues was also determined. Our work provided essential information for the function of ScPALs.

A new norsesquiterpenoid from Schisandra chinensis / 中国中药杂志

This study investigated the chemical components from the leaves and stems of Schisandra chinensis. Three norsesquiterpenoids were isolated from S. chinensis by various column chromatographies(silica gel, Sephadex LH-20, and MCI), reversed-phase medium-pressure preparative, and semi-preparative high-performance liquid chromatography(HPLC). Their structures were identified based on physicochemical properties, mass spectrometry(MS), nuclear magnetic resonance(NMR), ultraviolet(UV), and electro-nic circular dichroism(ECD) as(3R,4R,5R,6S,7E)-3,4,5,6-tetrahydroxy-7-megastigmen-9-one(1),(3S,5R,6R,7E)-3,5,6-trihydroxy-7-megastigmen-9-one(2), and(3S,4R,9R)-3,4,9-trihydroxymegastigman-5-ene(3). Compound 1 was a new compound, and its absolute configuration was determined by ECD. Compounds 2 and 3 were isolated from the Schisandra plant for the first time.

Mechanism analysis of Sanhan-Qushi-Wenjing-Tongluo formula treating psoas fasciitis based on biological information technology / 国际中医中药杂志

Objective:To explore the effective chemical constituents and target genes of the Sanhan-Qushi-Wenjing-Tongluo formula through the method of network pharmacology, and to further analyze the mechanism of treatoffing psoas fasciitis. Methods:The TCMSP database was used to search and screen the chemical active substances of Sanhan-Qushi-Wenjing-Tongluo formula and its target proteins acting on the human body. At the same time, the GeneCards database platform was used to predict the target of disease and the active ingredient-target network was constructed. Construct a PPI network through the STRING database, search for PPI core genes, and then perform GO enrichment analysis and KEGG enrichment analysis to find the signal pathways involved and construct a target-path network. Results:Through screening, a total of 23 key chemical components and 25 common target proteins was obtained in Sanhan-Qushi-Wenjing-Tongluo formula treating psoas fasciitis; gene analysis of enrichment analysis results include antibiotic response, cyclin-dependent proteins kinase holoenzyme complex, cytokine receptor binding, etc. Kyoto Encyclopedia of Genes and Genomes enrichment analysis results include AGE-RAGE signaling pathway, measles, endocrine resistance, inflammatory bowel disease, etc; the target genes gained which have a higher degree of matching with the above mentioned pathways include IL6, JUN, IL1B, CDK4, CCND1. Conclusion:Sanhan-Qushi-Wenjing-Tongluo formula could treat psoas fasciitis by regulating the target genes such as IL6, JUN, IL1B, CDK4 and CCND1.

Evaluating the long-term prognosis of coronary artery disease patients undergoing percutaneous coronary intervention by risk stratification with ACC/AHA classification of coronary lesions / 中华心血管病杂志

Objective@#To evaluate the long-term prognosis of coronary artery disease (CAD) patients undergoing percutaneous coronary intervention (PCI) by risk stratification with American College of Cardiology (ACC)/American Heart Association (AHA) classification of coronary lesions.@*Methods@#Data used in this study derived from the I-LOVE-IT 2 trial. I-LOVE-IT 2 trial was a prospective, multicenter, randomized, assessor-blinded, noninferiority study. A total of 1 255 patients in I-LOVE-IT 2 trial with only one lesion and underwent biodegradable polymer drug-eluting stent implantation were included and grouped according to ACC/AHA classification of coronary lesions, namely type A/B1 lesion group (n=184), type B2 lesion group (n=457) and type C lesion group (n=614). The primary endpoint was 48-month patient-oriented composite endpoint (PoCE), a composite of all-cause mortality, all myocardial infarction, stroke, and/or any revascularization. The secondary endpoints were target lesion failure (TLF), components of PoCE, major bleeding (bleeding academic research consortium(BARC) type 3-5) and definite/probable stent thrombosis within 48 months. The incidences of endpoint events were compared in the three groups. The multivariable Cox hazard ratio model was used to analyze the independent predictors of PoCE and TLF at 48 months.@*Results@#Incidences of PoCE at 48 months were significantly higher in patients with type C lesion compared with patients with type A/B1 (24.43%(150/614) vs. 14.13%(26/184), P<0.05) or B2 lesion (24.43%(150/614) vs. 15.97%(73/457), P<0.05). The multivariable Cox hazard ratio model showed that the type C lesion were the independent predictors of 48-month PoCE (HR=1.59, 95%CI 1.21-2.08, P<0.001) and TLF (HR=2.31, 95%CI 1.53-3.49, P<0.001). After multivariable adjustment, the HRs of PoCE for patients with type C lesion versus type A/B1 and type B2 were 1.91 (95%CI 1.25-2.92, P=0.003) and 1.64 (95%CI 1.23-2.20, P<0.001), respectively. Meanwhile, the HRs of TLF for patients with type C lesion versus type A/B1 and type B2 were 2.45 (95%CI 1.29-4.64, P=0.006) and 2.55 (95%CI 1.62-4.02, P=0.001), respectively.@*Conclusions@#The ACC/AHA classification of coronary lesions has good discrimination with long-term outcomes for CAD patients undergoing PCI. The type C lesion is associated with a worse prognosis, enough attention should be paid in these patients during routine clinical management.

Clinical observation on herb-partitioned spreading moxibustion at Baliao points plus climen for diminished ovarian reserve / 针灸推拿医学（英文版）

Objective: To observe the clinical efficacy of herb-partitioned spreading moxibustion at Baliao points plus climen for diminished ovarian reserve (DOR). Methods: A total of 60 patients with DOR were randomized into a spreading moxibustion group and a Western medicine group by the random number table method, with 30 cases in each group. The Western medicine group was treated with climen, starting from the 5th day of the menstrual cycle for 21 d. The spreading moxibustion group was treated with herb-partitioned spreading moxibustion at Baliao points on the basis of the medication in the Western medicine group, 1 h per time, once a week. The treatment was performed for 1 month as one treatment course in both groups, for 3 courses in total. The serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) in the patients were measured before and after treatment. The peak systolic velocity (PSV) and resistance index (RI) were also detected. The traditional Chinese medicine (TCM) symptom score was evaluated. The clinical efficacy was evaluated after treatment. Results: The total effective rate in the spreading moxibustion group was 93.3％, which was significantly higher than 80.0％ in the Western medicine group, and the difference between the groups was statistically significant (P<0.05). After treatment, the TCM symptom scores, the serum FSH levels, FSH/LH ratios and RI in both groups decreased, and the intra-group differences were all statistically significant (all P<0.05). The serum E2 level and PSV increased compared with those in the same group before treatment, and the intra-group differences were statistically significant (all P<0.05). After treatment, the TCM symptom score, the serum FSH level, FSH/LH ratio and RI in the spreading moxibustion group were lower than those in the Western medicine group, while the serum E2 level and PSV were higher than those in the Western medicine group, and the differences between the groups were statistically significant (all P<0.05). Conclusion: Herb-partitioned spreading moxibustion at Baliao points plus climen can produce valid therapeutic efficacy for DOR. It can improve the clinical symptoms, regulate serum hormone levels and increase ovarian blood perfusion, thus improving ovarian reserve function, producing more significant efficacy than climen alone.

Evaluating the long-term prognosis of coronary artery disease patients undergoing percutaneous coronary intervention by risk stratification with ACC/AHA classification of coronary lesions / 中华心血管病杂志

Objective: To evaluate the long-term prognosis of coronary artery disease (CAD) patients undergoing percutaneous coronary intervention (PCI) by risk stratification with American College of Cardiology (ACC)/American Heart Association (AHA) classification of coronary lesions. Methods: Data used in this study derived from the I-LOVE-IT 2 trial. I-LOVE-IT 2 trial was a prospective, multicenter, randomized, assessor-blinded, noninferiority study. A total of 1 255 patients in I-LOVE-IT 2 trial with only one lesion and underwent biodegradable polymer drug-eluting stent implantation were included and grouped according to ACC/AHA classification of coronary lesions, namely type A/B1 lesion group (n=184), type B2 lesion group (n=457) and type C lesion group (n=614). The primary endpoint was 48-month patient-oriented composite endpoint (PoCE), a composite of all-cause mortality, all myocardial infarction, stroke, and/or any revascularization. The secondary endpoints were target lesion failure (TLF), components of PoCE, major bleeding (bleeding academic research consortium(BARC) type 3-5) and definite/probable stent thrombosis within 48 months. The incidences of endpoint events were compared in the three groups. The multivariable Cox hazard ratio model was used to analyze the independent predictors of PoCE and TLF at 48 months. Results: Incidences of PoCE at 48 months were significantly higher in patients with type C lesion compared with patients with type A/B1 (24.43%(150/614) vs. 14.13%(26/184), P<0.05) or B2 lesion (24.43%(150/614) vs. 15.97%(73/457), P<0.05). The multivariable Cox hazard ratio model showed that the type C lesion were the independent predictors of 48-month PoCE (HR=1.59, 95%CI 1.21-2.08, P<0.001) and TLF (HR=2.31, 95%CI 1.53-3.49, P<0.001). After multivariable adjustment, the HRs of PoCE for patients with type C lesion versus type A/B1 and type B2 were 1.91 (95%CI 1.25-2.92, P=0.003) and 1.64 (95%CI 1.23-2.20, P<0.001), respectively. Meanwhile, the HRs of TLF for patients with type C lesion versus type A/B1 and type B2 were 2.45 (95%CI 1.29-4.64, P=0.006) and 2.55 (95%CI 1.62-4.02, P=0.001), respectively. Conclusions: The ACC/AHA classification of coronary lesions has good discrimination with long-term outcomes for CAD patients undergoing PCI. The type C lesion is associated with a worse prognosis, enough attention should be paid in these patients during routine clinical management.

Value of SYNTAX revascularization index on evaluating the long-term prognosis of coronary artery disease patients with biodegradable polymer drug-eluting stents implantation / 中华心血管病杂志

Objective@#To explore the value of SYNTAX revascularization index (SRI) on evaluating the long-term prognosis of coronary artery disease (CAD) patients implanted with biodegradable polymer drug-eluting stents (BP-DES) and define the best threshold of SRI for predicting all-cause mortality in these patients.@*Methods@#Data used in this study derived from the I-LOVE-IT 2 trial (evaluate safety and effectiveness of the Tivoli DES and the Firebird DES for treatment of coronary). I-LOVE-IT 2 trial was a prospective, multicenter, randomized, assessor-blinded, non-inferiority study. A total of 1 829 patients implanted with BP-DES were divided into 3 groups, namely SRI=100% group (n=963), 50%≤SRI<100% group (n=527) and SRI<50% group (n=339). The primary endpoint was 48-month patient-oriented composite endpoint (PoCE), a composite of all-cause mortality, myocardial infarction(MI), stroke, and/or any revascularization. The secondary endpoints were components of PoCE and definite/probable stent thrombosis at 48 months. The receiver operating characteristic curve was used to investigate the best cut-off point of SRI for 48-month all-cause mortality. The Cox regression analysis was used to identify independent predictors of the all-cause death and PoCE at 48 months.@*Results@#Incidence of PoCE at 48 months was significantly lower in SRI=100% group than patients with 50%≤SRI<100%(17.34% (167/963) vs. 22.20% (117/527), P<0.05) and SRI<50% (17.34% (167/963) vs. 24.78% (84/339), P<0.05). Comparing with SRI=100% group, the patients with 50%≤SRI<100% suffered higher rates of all MI (7.78% (41/527) vs. 4.26% (41/963), P<0.05) and target vessel MI (6.45% (34/527) vs. 4.26% (41/963), P<0.05); patients with SRI<50% had higher rates of all-cause mortality (5.90% (20/339) vs. 3.12% (30/963), P<0.05) and any revascularization (14.16% (48/339) vs. 3.12% (30/963), P<0.05). The receiver operating characteristic curve analysis showed that the SRI=65% was the best cut-off point to predict the all-cause mortality at 48 months (area under the curve was 0.58, sensitive was 0.47, specificity was 0.70). Meanwhile, SRI<65% was an independent predictor of 48-month all-cause mortality (HR=2.06, 95%CI 1.25-3.38) and PoCE (HR=1.34, 95%CI 1.09-1.66).@*Conclusions@#SRI serves as a good index for predicting long-term prognosis and SRI<65% is an independent predictor of 48-month PoCE and all-cause mortality for CAD patients with BP-DES implantation. Meanwhile, SRI≥65% might be a reasonable threshold of incomplete revascularization.

Lentivirus-mediated BMP-2 overexpression plasmid transfection into bone marrow mesenchymal stem cells combined with silk fibroin scaffold for osteoblast transformation / 中国骨伤

OBJECTIVE@#To explore the effect of lentivirus-mediated BMP-2 overexpression plasmid transfection into bone marrow mesenchymal stem cells and silk fibroin scaffold on osteoblast transformation.@*METHODS@#The lentivirus BMP-2 overexpression vector was constructed, bone marrow mesenchymal stem cells were cultured, and the combined culture system of nuclear scaffolds was constructed. Alizarin red staining and alkaline phosphatase staining were used to detect the osteogenic transformation of bone marrow mesenchymal stem cells in vitro. Ten New Zealand white rabbits, weighing 3.2 to 4.5 kg(averaging 3.9 kg), aged (2.89±0.45) years old, were selected to construct the rabbit tibial defect model by drilling a conical tibial defect (5 mm in length, 2 mm in width and 3 mm in depth) with an oral drill. The repair of the tibial defect in the animal model was observed by HE staining. The experimental group was implanted with silk fibroin scaffold + BMP-2 overexpression vector bone marrow mesenchymal stem cell complex, while the negative control group was implanted with silk fibroin scaffold+non-transfected bone marrow mesenchymal stem cell complex.@*RESULTS@#Compared with the control group(silk fibroin scaffold+non-transfected bone marrow mesenchymal stem cells), the number of adherent cells on the surface of the scaffold in the experimental group(silk fibroin scaffold+transfected BMP-2 overexpression vector BMP-2 complex) increased significantly. Compared with the control group, the ECM secretion in the experimental group increased significantly. EDX analysis showed that the content of calcium ion was 0.22% in the control group and 0.86% in the experimental group, which showed that the ability of inducing calcium ion formation in the experimental group was stronger than that in the control group. Alizarin red staining of calcium nodules showed that there was no obvious change in the naked eye of the control group, and a small amount of calcium nodules could be seen under the microscope. In the experimental group, obvious red area staining was observed by naked eye, and a large number of calcium nodules were observed by microscopy. The results of alkaline phosphatase staining showed that there was no obvious change in the naked eye of the control group, and no obvious change in the microscopic observation. In the experimental group, purple area staining was observed by naked eyes, and ALP staining was strongly positive by microscopy. The combined culture system of silk fibroin scaffold and bone marrow mesenchymal stem cells can repair cartilage defects. The repair effect of BMP-2 bone marrow mesenchymal stem cells after transfection is obviously better than that of non-transfection group. HE staining showed that inflammatory cells decreased and scaffolds disappeared slightly in the control group. In the experimental group, inflammatory cells were significantly reduced, scaffolds disappeared and angiogenesis was observed.@*CONCLUSIONS@#Lentivirus-mediated BMP-2 overexpression plasmid can promote BMSC to differentiate into osteocytes and secrete more extracellular matrix containing Ca²⁺ to promote bone defect repair.

A novel phenotype with splicing mutation identified in a Chinese family with desminopathy / 中华医学杂志(英文版)

BACKGROUND@#Desminopathy, a hereditary myofibrillar myopathy, mainly results from the desmin gene (DES) mutations. Desminopathy involves various phenotypes, mainly including different cardiomyopathies, skeletal myopathy, and arrhythmia. Combined with genotype, it helps us precisely diagnose and treat for desminopathy.@*METHODS@#Sanger sequencing was used to characterize DES variation, and then a minigene assay was used to verify the effect of splice-site mutation on pre-mRNA splicing. Phenotypes were analyzed based on clinical characteristics associated with desminopathy.@*RESULTS@#A splicing mutation (c.735+1G>T) in DES was detected in the proband. A minigene assay revealed skipping of the whole exon 3 and transcription of abnormal pre-mRNA lacking 32 codons. Another affected family member who carried the identical mutation, was identified with a novel phenotype of desminopathy, non-compaction of ventricular myocardium. There were 2 different phenotypes varied in cardiomyopathy and skeletal myopathy among the 2 patients, but no significant correlation between genotype and phenotype was identified.@*CONCLUSIONS@#We reported a novel phenotype with a splicing mutation in DES, enlarging the spectrum of phenotype in desminopathy. Molecular studies of desminopathy should promote our understanding of its pathogenesis and provide a precise molecular diagnosis of this disorder, facilitating clinical prevention and treatment at an early stage.

Clinical observation on acupoint injection of Chuankezhi injection plus acupoint sticking in treating bronchial asthma / 针灸推拿医学（英文版）

Objective: To observe the clinical efficacy of acupoint injection of Chuankezhi plus acupoint sticking treatment for bronchial asthma, and provide clinical evidence for the synergy of the two therapies, and explore their synergistic mechanism. Methods: A total of 70 patients were randomized into an acupoint injection plus sticking group and an acupiont sticking group by the random number table, with 35 cases in each group. The treatment took place in July and August. The acupoint injection plus sticking group was treated with acupoint injection and acupiont sticking, while the acupiont sticking group was treated only with acupiont sticking therapy. The treatment course was 4 weeks. After the treatment, the scores of symptom scale in the two groups before treatment, 3 months and 6 months after the treatment were observed. Results: During the treatment, there were 5 dropouts in the acupoint injection plus sticking group with 30 cases remained, and 4 dropouts in the acupiont sticking group with 31 cases remained. Before the treatment, there was no significant difference in the total scores of symptom scale between the two groups. Three months after the treatment, the total scores of symptom scale of both groups were lower than those before treatment, and the intra-group differences were statistically significant (bothP<0.05). In the inter-group comparison, there was no significant difference in the difference values of the scores before treatment and 3 months after treatment (P>0.05). Six months after the treatment, the total scores of symptom scale in both groups were lower than those before treatment, and the intra-group differences were statistically significant (bothP<0.05), so were the differences in the difference values of the scores before treatment and 6 months after treatment (bothP<0.05). Conclusion: Acupoint injection of Chuankezhi plus acupoint sticking or acupoint sticking alone both can improve the symptoms of patients with bronchial asthma. Acupoint injection of Chuankezhi plus acupoint sticking has a higher total effective rate than single acupoint sticking therapy.

Evaluation of progression-free survival for locally advanced rectal cancer by MRI after neoadjuvant chemoradiotherapy and total mesorectal excision / 中华肿瘤杂志

Objective@#To evaluate the predictive value of MRI features and pathological parameters on local recurrence, metastasis and progression free survival (PFS) for locally advanced rectal cancer after neoadjuvant chemoradiotherapy and subsequent total mesorectal excision surgery.@*Methods@#A retrospective analysis of 95 patients with locally advanced rectal adenocarcinoma who underwent total mesorectal excision after neoadjuvant chemoradiotherapy was performed. Univariate and multivariate analyses were performed to evaluate the predictive value of MRI features before chemoradiation and postoperative pathological parameters on progression free survival.@*Results@#Among the 95 cases, 5 cases occured local recurrence, 21 cases developed, 3 cases including both locally recurrence and distant metastasis, 19 died and 47 had no recurrence or metastasis at the last of follow-up. Univariant analysis showed that MRI signs before chemoradiation, namely, mr circumferential resection margin, mr levator ani muscle invasion, mr lymphatic vessel invasion, mr tumor deposition and postoperative pathological parameters, yp circumferential resection margin, yp lymphatic vessel invasion were related to PFS (P<0.05). Multivariate analysis of Cox proportional hazard model showed that mr lymphatic vessel invasion and mr tumor deposition were independent factors for PFS (OR=2.774 and 3.029, P<0.05).@*Conclusions@#Lymphatic vessel invasion and tumor deposition on MRI are independent prognostic factors for progression free survival of locally advanced rectal cancer after neoadjuvant chemoradiotherapy and TME surgery. To some extent, MRI signs can assess local recurrence and distant metastasis in locally advanced rectal cancer patients after neoadjuvant chemoradiotherapy and mesorectal excision.

Phenolic acid derivatives of Euphorbia ebracteolata / 中国中药杂志

A variety of chromatographic methods were applied to research the chemical composition of the roots of Euphorbia ebracteolata. Meanwhile, the structures of these compounds were identified by spectroscopic data, such as 1D, 2D NMR and HR-ESI-MS. In the present research, a new phenolic acid compound, 1,3,4,5-tetrahydroxy-2-O-β-D-[(6'-Gallic acyl)-glucopyranyl] isopentane(1), was isolated from the roots of E. ebracteolata, as well as three known compounds, isopropyl-2-O-β-D-(6'-gallic acyl)-glucopyranoside(2), ellagic acid(3) and 3,4'-di-O-methylellagic acid(4) respectively. Additionally, compounds 2 and 4 were isolated from the Euphorbia genus for the first time.

Construction and inhibitory effect of microRNA expression vector targeting survivin on proliferation of human colorectal carcinoma HT-29 cells / 国际肿瘤学杂志

Objective To construct microRNA (miRNA) expression vector targeting surviving,and to investigate its effect on transfected human colorectal carcinoma (HT-29) cell apoptosis and proliferation.Methods miRNA targeting survivin was synthesized and transfected HT-29 cells by lipofectin.HT-29 cells were cultured in the 6 orifices.The cultured cells were divided into control,liposome,negative control and positive control groups.Transient transfected cells were collected and the proliferation index and apoptosis rate of HT-29 cells were detected by flow cytometry.The expressions of survivin mRNA and protein were detected by RT-PCR and Western blot.Results The proliferation index and apoptosis rate of the positive control group were significantly higher compared with normal group,transfection group and mock-vehicle group (17.98％ ± 2.35％ vs 38.04％ ±2.11％ vs 36.73％ ±2.51％ vs 36.57％ ±3.05％; t =20.05,P＜0.01; t =18.75,P＜0.01; t=18.59,P＜0.01; 19.54％ ±1.74％ vs 3.13％ ±0.29％ vs 3.70％ ±0.44％ vs 3.61％ ± 0.50％ ; t =16.40,P ＜ 0.01 ; t =15.84,P ＜ 0.01 ; t =15.92,P ＜ 0.01).Survivin mRNA and protein expression levels were specifically suppressed in transfected HT-29 cells (t =0.68,P ＜0.01 ; t =0.58,P ＜ 0.01; t=0.61,P＜0.01;t=0.64,P＜0.01; t=0.62,P＜0.01;t=0.67,P＜0.01).Conclusion Survivin targeted silence can effectively decrease the expression of survivin mRNA and protein,induce colorectal carcinoma HT-29 cell apoptosis and inhibit cell proliferation.

Effects of moxibustion on concentration of extracellular potassium ion in acupoint under different status / 中国针灸

<p><b>OBJECTIVE</b>To observe different effects of moxibustion on extracellular potassium ion in acupoint under physiological and pathological status and provide experimental evidence for exploring action mechanism of moxibustion on acupoint local.</p><p><b>METHODS</b>Forty female SD rats were randomly divided into a blank group, a blank-moxibustion group, a model group and a model-moxibustion group, 10 cases in each one. The complete Freund's adjuvant(CFA) was adopted to establish model of adjuvant arthritis (AA) in the model group and model-moxibustion group. No treatment was given in the blank group and model group while moxibustion was applied at "Zusan-li" (ST 36) for 30 min in the blank-moxibustion group and model-moxibustion group. The tissue fluid in "Zusanli" (ST 36) was collected with microdialysis and real-time analyzed by electrolytic analyzer. The change of concentration of potassium ion in "Zusanli" (ST 36) was observed.</p><p><b>RESULTS</b>(1) Under physiological status, the concentration of extracellular potassium ion in the blank group was not changed within 150 min (P > 0.05); before the moxibustion, the concentration of extracellular potassium ion in the blank-moxibustion group was (1.21 +/- 0.31) mmol/L, and after treatment it was gradually increased and reached its peak at (2.38 +/- 0.42) mmol/L after 60 min (P < 0.05), then it was reduced. 150 min after the treatment, concentration of potassium ion was slightly higher than that before moxibustion as well as that in the blank group. The concentration in the blank-moxibustion group at 60 min was statistically significant compared with that in the blank group (P < 0.05). (2) Under pathological status, the concentration of extracellular potassium ion in the model group was not changed within 150 min, differences of which at each time point was not statistically significant (all P > 0.05). Before the moxibustion, the concentration of extracellular potassium ion was (1.09 +/- 0.12) mmol/L in the model-moxibustion group, and it was immediately increased to (1.96 +/- 0.18) mmol/L after moxibustion. 60 min and 90 min after the moxibustion, it still maintained a higher level, which was (1.87 +/- 0.29) mmol/L and (1.59 +/- 0.16) mmol/L respectively (both P < 0.05). The differences of each time point after moxibustion in the model-moxibustion group were statistically significant compared with those in the model group (all P < 0.05).</p><p><b>CONCLUSION</b>The moxibustion could increase the concentration of potassium ion in rat's acupoint local under physiological status but time of effect is short; with moxibustion at "Zusanli" (ST 36) under pathological status, the concentration of local potassium ion is obviously increased and maintains for a long time.</p>

Pathological changes in the epileptogenic foci of children with intractable epilepsy / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate pathological changes in the epileptogenic foci of children with intractable epilepsy and their clinical significance.</p><p><b>METHODS</b>Thirty children with intractable epilepsy were included in the study. The epileptogenic foci were surgically resected and pathological changes in the obtained specimens were observed under a light microscope (LM) and a transmission electron microscope (TEM).</p><p><b>RESULTS</b>Under the LM, cortical dysplasia was found in 14 cases (47%), hippocampal sclerosis in 11 cases (37%), dysembryoplastic neuroepithelial tumor in 1 case (3%), ganglioglioma in 1 case (3%), and encephalomalacia in 3 cases (10%). The TEM observation revealed pathological changes in the ultrastructure of the hippocampus and extra-hippocampal cortex, such as changes in the number of synapses and synaptic structure, decrease in neurons and karyopyknosis, swelling and degeneration of astrocytes, and changes in mitochondrial structures.</p><p><b>CONCLUSIONS</b>Pathological changes in the hippocampus and extra-hippocampal cortex, especially synaptic remodeling, may be the morphological basis for spontaneous recurrent seizures in children with intractable epilepsy. The pathological changes and epileptiform activity are related to an imbalance between excitatory and inhibitory neurotransmission.</p>

Sonic hedgehog signaling enhanced the expression of histone demethylase, lysine-specific demethylase 8 in the head and neck squamous cell carcinoma cell line SCC-6 / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To determine whether the sonic hedgehog (Shh) signaling could regulate the expression of histone demethylases in the head and neck squamous cell carcinoma(SCC).</p><p><b>METHODS</b>Human recombinant SHH-N protein or over-expression of the mutant 2 smoothened (M2-SMO) was applied to activate the Shh signaling in tongue squamous cell carcinoma cell line-SCC-6 in this study. Cyclopamine was used to block the Shh signaling in SCC-6. The real-time reverse transcription (RT)-PCR was used to detect the expression of histone demethylases at the mRNA level.</p><p><b>RESULTS</b>The data showed that activation of the Shh signaling up-regulated the expression of histone demethylase, lysine-specific demethylase 8 (KDM-8) at the mRNA level by human recombinant SHH-N protein (1.841 ∼ 3.591 fold compare with untreated group; P < 0.01), over-expression of the M2-SMO also increased the expression of KDM-8 (1.358 ∼ 3.013 fold compared with empty vector group; P < 0.05), and after the Shh signaling was blocked by Cyclopamine, the expression of KDM-8 was down regulated (decreased 25.6% ∼ 66.6% compared with control cells, P < 0.05).</p><p><b>CONCLUSIONS</b>Histone demethylase KDM-8 was downstream target gene of Shh signaling in head and neck squamous cell carcinoma cell line SCC-6, and its expression was positively regulated by the Shh signaling.</p>

Preparation condition optimization of astragalus polysaccharide liposome by orthogonal test / 中草药

Objective: To optimize the preparation conditions of astragalus polysaccharide (APS) liposome. Methods: APS liposome was prepared by membrane distribution-micromembrane extruding method. L9(34) orthogonal test was adopted to optimize the preparation conditions which was designed with three factors of lecithin to drug ratio, lecithin to cholesterol ratio, and ultrasonic time, and two indexes of encapsulation efficiency and drug-loading rate. The encapsulation efficiency of APS liposome was assayed by Protamine method, the shape and particle size were observed by transmission electron microscope. Results: The optimized preparation conditions were as follows: the ratio of lecithin to drug was 10:1, the ratio of lecithin to cholesterol was 8:1, and ultrasonic time was 20 min. Conclusion: The APS liposome prepared under the optimized conditions has following advantages: high encapsulation efficiency and drug-loading rate, uniform shape and particle size, and good reproduction quality.

Effects of different anesthetic and analgesic protocols on cellular immune function and stress hormone level in patients undergoing lobectomy for esophagus cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of different anesthetic and analgesic protocols on the cellular immune function and stress hormone in patients undergoing lobectomy for esophagus cancer.</p><p><b>METHODS</b>Sixty ASA I or II patients undergoing lobectomy for esophagus cancer were randomly divided into two groups to receive postoperative general anesthesia and intravenous analgesia (group A, n=30) or intraoperative general anesthesia combined with thoracic epidural anesthesia with postoperative epidural analgesia (group B, n=30). The cervical venous blood samples were obtained from the patients at 30 min before anesthesia induction (T(0)), 2 h after skin incision (T(1)), and at 4 h (T(2)), 24 h (T(3)) and 48 h (T(4)) after the end of operation. The T-lymphocyte subsets (CD4(+) and CD8(+)) were analyzed by flow cytometry, serum concentrations of sIL-2R and IL-2 determined by ELISA, and the levels of growth hormone (GR), prolactin (PRL), IL-8 and cortisol (Cor) measured by radioimmunoassay. Visual analogue scale (VAS) was used for assessment of the postoperative analgesic effects.</p><p><b>RESULTS</b>The VAS scores were significantly lower in group B than in group A at T(2) and T(3) (P<0.05). The percentage of CD4(+) cells and the CD4(+)/CD8(+) ratio in the two groups began to decrease significantly at T(1) (P<0.05), reducing to the lowest level at T(2) in group B and at T(3) in group A. From T(1) to T(4), the percentage of CD4(+) in group B remained significantly higher than those in group A (P<0.05), and from T(3) to T(4), the CD4(+)/CD8(+) ratio in group B were significantly higher than those in group A (P<0.05). The IL-2 level in the two groups began to decrease significantly at T(1) (P<0.05), reaching the lowest level at T(2) in group A and at T(3) in group A. IL-2 level was significantly higher in group B than in group A from T(3) to T(4) (P<0.05). sIL-2R level in group A began to increase at T(1) and peaked at T(3), showing significant differences from the T(0) level, but the level showed no significant variations in group B compared with the T(0) level. From T(2) to T(4), sIL-2R level was significantly higher in group A than in group B (P<0.05). The levels of GH, PRL and Cor increased significantly, while IL-8 decreased in the two groups from T(1) to T(4) (P<0.05), but remained stable in group B.</p><p><b>CONCLUSION</b>General anesthesia combined with thoracic epidural anesthesia may reduce the perioperative stress reaction and adverse effect on cellular immune function in patients undergoing lobectomy for esophagus cancer.</p>

Increase of saliva nitrate and nitrite level in patients with oral candidiasis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To observe the alterations of saliva nitrate and nitrite level in patients with oral candidiasis.</p><p><b>METHODS</b>Parotid saliva and whole saliva were collected from 33 patients and 34 healthy volunteers. Concentrations of nitrate and nitrite in saliva were determined by high-performance liquid chromatography. Follow-up observation was performed on 10 patients after treatment. The data were statistically analyzed with independent-samples t test or paired-samples t test at alpha = 0.05.</p><p><b>RESULTS</b>There was significant increase of the concentrations and secretion rate of parotid saliva nitrate in patient group as compared with controls: (49.70 +/- 0.50) vs (21.51 +/- 0.60) mg/L (t = 2.692, P = 0.009) and (27.71 +/- 0.50) vs (12.55 +/- 0.60) microg/min (t = 2.554, P = 0.013), respectively. Significantly increased concentrations and secretion rate of nitrate and nitrite [nitrate: (6.46 +/- 0.94) vs (1.11 +/- 0.70) mg/L (t = 3.792, P = 0.000); nitrite: (8.48 +/- 0.58) vs (3.39 +/- 0.53) mg/L (t = 2.888, P = 0.005); nitrate secretion rate: (10.57 +/- 0.91) vs (2.10 +/- 0.74) microg/min (t = 3.464, P= 0.001); nitrite secretion rate: (13.91 +/- 0.55) vs (6.42 +/- 0.58) microg/min (t = 2.397, P = 0.020)] were revealed in whole saliva of patients group. Significantly decreased nitrate and nitrite levels were also observed in patients after treatment, especially the changes of parotid saliva nitrate secretion rate [(37.50 +/- 0.50) vs (14.34 +/- 0.64) microg/min (t = 3.142, P = 0.012)], whole saliva nitrate [(14.29 +/- 1.01) vs (2.59 +/- 1.03) mg/L (t = 3.475, P = 0.007)] and whole saliva nitrate secretion rate [(25.97 +/- 0.93) vs (4.12 +/- 1.00) microg/min (t = 3.922, P = 0.003)].</p><p><b>CONCLUSION</b>The present study revealed the significant increase of salivary nitrate and nitrite level in patients with oral candidiasis is considered to be associated with the host defense reaction.</p>

Study on the relationship of serum vitamin E and liver pathological features in the patients with chronic hepatitis B / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study on the relationship of serum vitamin E and liver pathological features in the patients with chronic hepatitis B.</p><p><b>METHODS</b>Sixty-six patients with chronic hepatitis B and ten healthy controls were enrolled in this present study. The serum vitamin E level was measured spectrophotometrically. Comparisons of liver function test, HBeAg and HBV DNA level were conducted among different liver pathological features including inflammatory grading and fibrosis staging.</p><p><b>RESULTS</b>Compared with healthy controls, the serum level of vitamin E was significantly decreased in the patients with chronic hepatitis B, especially in those with elevated ALT activity. In comparison between HBeAg positive group and HBeAg negative group, the serum level of vitamin E of the former group did not significantly changed (P > 0.05). Furthermore, the serum level of vitamin E has been demonstrated to be negatively associated with the inflammation grading in the patients with chronic hepatitis B. However, there was no significant association between the serum vitamin E and liver fibrosis staging.</p><p><b>CONCLUSION</b>Vitamin E, as one of the important anti-oxidants, was demonstrated to be implicated in the progression of liver inflammation in the patients with chronic hepatitis B. Furthermore, the supplement of vitamin E would be a potential therapy for attenuate the inflammatory response.</p>