OBJECTIVETo investigate molding technology of total alkaloids from Sophora alopecuroides freeze-dried powders and observe its inhibition effects on liver transplantation tumor in mice.

METHODWith color, clarity, water-soluble and formability as indexes, single factor tests were adopted to screen type and amount of filler, the concentration of total alkaloids in drug liquid, pH in order to determine optimum prescription of total alkaloids from S. alopecuroides freeze-dried powders, the lowest melting point was determined and freeze drying curve was drafted. Mice hepatoma H22 ascites tumor strain was sterile inoculated in right axillary subcutaneous of mice, and antitumor effect of total alkaloids from S. alopecuroides freeze-dried powders on liver transplantation tumor H22 in mice.

RESULTWhen selected 80 g x L(-1) as mannitol as filler, the concentration of total alkaloids in drug liquid was 25 g x L(-1) and pH 6.5-7.5, freeze-dried effect was optimum with fast reconstitute speed. Average inhibition rate of the big (120 mg x kg(-1)) and medium (60 mg x kg(-1)) dose group of total alkaloids from S. alopecuroides freeze-dried powders on liver transplantation tumor H22 in mice were 56.08% and 35.49%, respectively.

CONCLUSIONPreparation technology was reasonable, reproducible and stable, total alkaloids from S. alopecuroides freeze-dried powders had significant antitumor effect and showed a dose-effect relationship.

Alkaloids ; chemistry ; pharmacology ; Animals ; Antineoplastic Agents, Phytogenic ; chemistry ; pharmacology ; Cell Line, Tumor ; Color ; Freeze Drying ; Humans ; Liver Neoplasms ; drug therapy ; Mice ; Powders ; chemistry ; Sophora ; chemistry ; Tumor Burden ; drug effects

Pseudorabies virus (PRV) is a swine herpesvirus of the Alphaherpesvirinae subfamily and a pathogen of swine resulting in devastating disease and economic losses worldwide. Cre/loxP site-specific system has the character of site specific, time specific, tissue specific and high efficiency in recombination, which makes this system universal in vivo and in vitro recombination of bacteria, fungus, plants, insects and mammals. A recombinant PRV which contain a loxP site in TK locus by using Cre/LoxP recombinant system was construsted. A pair of primers were synthesized according to the pEGFP-C1 sequence published on GenBank, and were used to amplify the EGFP gene expression cassette with two loxP sites flanking each side. This target gene was cloned into pSKLR, the resulting transfer vector pSKLR-GFP-loxP was then cotransfected into 293T cells with PRV SH strain genomic DNA. The recombinant virus rPRV1 was selected and purified in TK-143 cells by choosing fluorescent expressing plaques. Cre expression vector pOG231 was cotransfected into 293T cells with rPRV1 genomic DNA. The second recombinant virus rPRV2 was obtained, which contains only one loxP site in TK locus. Sequencing results of rPRV2 TK gene indicated that 34bp loxP site was inserted into rPRV2 genome and there were 270bp deletion in TK gene. PCR amplifying different generations of rPRV2 TK gene showed that the mutant was stable when passages in RK-13 cells. TCID_ 50 assay indicated that rPRV2 grows well on RK-13 cells. The LD_ 50 test results on BALB/C mice suggested that the virulence of rPRV2 was reduced. As a conclusion, the report gene GFP expression cassette was removed successfully from rPRV1 genome and only one LoxP site was leaved in rPRV2 genome by using Cre/LoxP recombinant system.

OBJECTIVETo observe the differences in onset time and the overall efficacy of the acupuncture therapy of dredging Governor Vessel and regulating mentality combined with fluoxetine and the fluoxetine for post-stroke depression(PSD).

METHODSSixty-three patients were randomly divided into an acupuncture intervention of dredging Governor Vessel and regulating mentality group(acupuncture and medication group, 33 cases) and a control group (medication group, 30 cases). In the medication group, 20 mg fluoxetine was used by oral administration, once a day at 7:00 in the morning, continuously for 4 weeks. In the acupuncture and medication group, based on the oral administration of fluoxetine, the acupuncture intervention of dredging Governor Vessel and regulating mentality was applied mainly at Baihui(GV 20), Fengfu(GV 16), Shenting(GV 24), Shuigou(GV 26), Dazhui(GV 14) and Shendao(GV 11), once a day, 6 times a week, continuously for 4 weeks. Twenty-four items in Hamilton Depression Scale(HAMD), modified Edinburgh Scandinavia Stroke Scale(MESSS) and activity of daily life scale(ADL, Barthel index, BI) were used before and after 2-week and 4-week treatment. The efficacy was compared between the two groups.

RESULTSAfter 2-week treatment, the HAMD score and the MESSS score in the acupuncture and medication group were obviously decreased and the BI score was apparently increased(all P<0. 01). In the medication group, however, the score of every scale was not statistically different from that before treatment(P'>0. 05). After 4-week treatment, HAMD scores and MESSS scores in the two groups were obviously decreased and the BI scores I were apparently increased(all P<0. 01). After 2-week and 4-week treatment, the HAMD scores and the MESSS scores in the acupuncture and medication group were lower than those in the medication group and the BI scores were higher than those in the medication group(P<0. 01, P<0. 05). The total effective rate of anti-depression (97. 0%, 32/33) the total effective rate of nerve function impairment(90. 9%, 30/33) and the total effective rate of daily life activity(97. 0% 32/33) in the acupuncture and medication group were better than 80. 0% (24/30), 80. 0%(24/30), 83. 3%(25/30) in the medication group(all P<0. 05).

CONCLUSIONThe acupuncture therapy of dredging Governor Vessel and regulating mentality could reduce the onset time of anti-depression medicine treatment of PSD and enhance the overall efficacy. Therefore, it enhances the clinical compliance.

Acupuncture Therapy ; Adult ; Aged ; Antidepressive Agents ; administration & dosage ; Combined Modality Therapy ; Depression ; drug therapy ; etiology ; therapy ; Female ; Humans ; Male ; Meridians ; Middle Aged ; Stroke ; complications ; psychology

OBJECTIVETo observe the effects of Ginkgo Biloba Tablet (GBT) in treating mild cognitive impairment (MCI).

METHODSTotally 113 MCI patients including 58 outpatients at the cognitive impairment clinics of Huadong Hospital, 34 outpatients at Chinese Medicine Clinics of Dongfang Hospital, and 21 outpatients at Feng-lin Community Health Service Center, were assigned to the control group (55 cases) and the treatment group (58 cases). Basic treatment was given to them all. Those in the treatment group took GBT additionally, 1 tablet each time, 3 times a day for 12 successive months. The mini-mental state examination (MMSE) integral, clock drawing task (CDT) integral, MMSE efficacy, CDT efficacy, and dementia conversion rate were detected before treatment, half a year after treatment, and one year after treatment.

RESULTSThere was no significant difference in scores of MMSE and CDT integrals between the two groups before treatment (P>0.05). Compared with before treatment, each score in the treatment group remarkably increased in the treatment group at the 6 months and 12 months after treatment (P<0.01). They were higher than those of the control group at the same time points, showing statistical difference (P<0.01, P<0.05). There was no significant difference in the integrals of MMSE and CDT between the 6 months of treatment and the 12 months of treatment (P>0.05). The total effective rate of MMSE and CDT of the treatment group at the 6 months and 12 months after treatment (MMSE 67.24% and 65.52%; CDT 62.07% and 60.34%) all exceeded those of the control group at the same time points (MMSE 36.36% and 30.91%; CDT 34.55% and 30.91%), showing significant difference (P<0.01). There was no difference in the total effective rate of MMSE and CDT of the treatment group between the two time points (P>0.05). The dementia conversion rates at the 6 months and 12 months after treatment (1.72% and 5.17%) were lower in the treatment group than in the control group (9.09% and 14.55%), showing statistical difference (P<0.05).

CONCLUSIONSOne-year GBT could significantly improve the cognitive function of MCI patients. It could also lower the dementia conversion rate.

Aged ; Aged, 80 and over ; Cognition ; Cognitive Dysfunction ; drug therapy ; psychology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Flavones ; pharmacology ; therapeutic use ; Ginkgo biloba ; Humans ; Male ; Middle Aged ; Neuropsychological Tests ; Phytotherapy ; Tablets

OBJECTIVETo observe the effect of Ginkgo Leaves Tablet (GLT) on memory quotient (MQ) of mild cognitive impairment (MCI) patients.

METHODSOne hundred and thirteen patients were randomly assigned to the control group (55 cases) and the treatment group (58 cases). Patients in the control group received dietetic therapy and physical exercises, while those in the treatment group additionally took GLT, 19.2 mg each time, three times daily. The treatment course was 12 months for all. The MQ of all the patients was assessed by WMS-RC before treatment,at 6-month of treatment, and 12-month of treatment.

RESULTSCompared with the control group, the improvement of MQ increased in the treatment group 0.5 and 1 year after treatment (P < 0.05). The clinical efficiency of MQ obviously increased in the treatment group (48.28% and 50.00%), showing statistical difference when compared with the control group (30.91% and 27.27%, P < 0.05, P < 0.01). There was statistical difference in added scores of recognition, regeneration, understanding, and recitation test at 6-month of treatment and 12-month of treatment between the treatment group and the control group (P < 0.05, P < 0.01).

CONCLUSIONGLT was effective in improving MQ of MCI patients, especially in improving recognition, regeneration, understanding, and recitation test.

Aged ; Aged, 80 and over ; Cognitive Dysfunction ; drug therapy ; psychology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Ginkgo biloba ; chemistry ; Humans ; Male ; Memory ; Middle Aged ; Phytotherapy

OBJECTIVETo observe the effect of the combination of sub-MIC sodium houttuyfonate and erythromycin on biofilm of Staphylococcus epidermidis.

METHODThe serial dilution method was adopted to determine MIC of the combination of sodium houttuyfonate and erythromycin on S. epidermidis; the checkerboard method was used to evaluate the combination of sodium houttuyfonate and erythromycin on suspended bacteria of S. epidermidis; S. epidermidis biofilm was built in vitro, and XTT reduction assay was used to evaluate the effect of the combination of sub-MIC sodium houttuyfonate and erythromycin on the adhesion of S. epidermidis and bacterial metabolism inside the biofilm. Microscope was applied to observe the impact the single administration and combination of the two medicines under sub-MIC on biofilm morphology of S. epidermidis.

RESULTThe MIC of sodium houttuyfonate and erythromycin were 62.5, 7.812 5 mg x L(-1), respectively. The combination of 1/8MIC sodium houttuyfonate and 1/2MIC erythronmycin showed a synergistic effect on S. epidermidis. Sodium houttuyfonate, erythromycin and their combination had an inhibitory effect on the adhesion and metabolism of S. epidermidis biofilm bacteria, and made impact on the morphology of S. epidermidis biofilm.

CONCLUSIONThe sub-MIC sodium houttuyfonate and erythromycin have an inhibitory effect on S. epidermidis biofilm. The combination of sodium houttuyfonate and erythromycin shows a synergistic effect in inhibiting suspended bacteria and biofilm of S. epidermidis, particularly in inhibiting the metabolism of S. epidermidis biofilm bacteria and impacting the morphology of biofilm.

Alkanes ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Bacterial Adhesion ; drug effects ; Biofilms ; drug effects ; growth & development ; Dose-Response Relationship, Drug ; Drug Interactions ; Erythromycin ; pharmacology ; Microbial Sensitivity Tests ; Staphylococcus epidermidis ; drug effects ; physiology ; Sulfites ; pharmacology

OBJECTIVEThe structure-activity relationship between traditional Chinese medicine (TCM) compounds and antibacterial activity was studied by chemoinformatics approach.

METHODCytoscape and its plug-in ChemViz were applied to compute the 2D chemical structure similarity and topological parameter TPSA (topological molecular polar surface area), which measures cell permeability of chemicals, between TCM compounds and clinical antibacterials. The overall degree of structure similarity was then calculated and represented by E-value for the eight categories of TCM compounds and the known antibacterials.

RESULTOur results indicated that flavonoids showed good structural similarity with antibacterials and appropriate cell permeability, compared with those of the TCM compounds of the other categories. As flavonoids were featured by good drug safety, it suggested that they can be regarded as the preferred lead compounds skeleton structure source for further antibacterials synthesis.

CONCLUSIONThe application of chemoinformatics helps explore the structure-activity relationship between TCM compounds and the antibacterial activity and search for suitable antibacterial lead compounds skeleton structure source.

Anti-Bacterial Agents ; chemistry ; pharmacology ; Informatics ; methods ; Medicine, Chinese Traditional ; Software ; Statistics as Topic ; Structure-Activity Relationship

Objective To explore the relationship of polymorphisms of D17S1878, D17S932 sites and essential hypertension.Methods Sixty-seven pedigrees were collected at the region with high prevalence of hypertension. The polymorphisms of D17S1878 and D17S932 sites were genotyped using Genetic Analyzer and GeneScan Software. Case-control study in sibs with different phenotype was carried out and logistic analysis was used for multivariate analysis. Results There were significant differences on the distributions of age, male, drinking, average systolic pressure, average diastolic pressure, the characteristics of rash, body mass index (BMI), total cholesterol amount, triglyceride, low density lipoprotein(LDL) between the hypertensive-affected sibs and the normotensive sibs (P＜0.05). There was significant difference between the affected hypertensive and normotensive sibs in the D17S1878 site (P＜0.05), while there was no significant difference in D17S932 (P＞0.05 ). After non-conditional logistic analysis, data showed that both sites were not included in the model, while age( OR = 1.044,95%CI:1. 019-1. 069), drinking ( OR = 2. 644,95% CI : 1. 778-3. 932), the characteristics of rash ( OR = 3. 078,95%CI:1.721-5.504), triglyceride (OR= 1.305,95%CI: 1.016-1.676), LDL-C (OR= 1.787,95% CI:1. 296-2. 646), as risk factors, were included in the model. Conclusion The polymorphisms of D17S1878 and D17S932 possibly were not associated with essential hypertension.

The hypertension is one of chronic vascular diseases, which often implicates multiple tissues causing stroke, cardiac hypertrophy, and renal failure. A growing body of evidence suggests that inflammatory mechanisms are important participants in the pathophysiology of hypertension. In this study, the inflammatory status of these tissues (kidney, liver, heart, and brain) in spontaneously hypertensive rats (SHR) was analyzed and its molecular mechanism was explored. The tissues were dissected from SHR and age-matched control Wistar-Kyoto (WKY) rats to investigate the abundance of inflammation-related mediators (IL-1beta, TNFalpha, ICAM-1, iNOS, C/EBPdelta and PPARgamma). mRNA levels were determined by reverse transcription-polymerase chain reaction and protein expression was evaluated by Western blot. To evaluate the oxidative stress of tissues, carbonyl protein content and total antioxidant capacity of tissues were detected by spectrophotometry and ferric reduction ability power (FRAP) method. The results suggest that: (1) Expressions of inflammation-related mediators (IL-1beta, TNFalpha, ICAM-1, iNOS, C/EBPdelta and PPARgamma) in SHR were higher compared with those in WKY rats except no evident increase of IL-1beta mRNA in liver and brain in SHR. (2) Tissues in SHR contained obviously increased carbonyl protein (nmol/mg protein) compared to that in WKY rats (8.93+/-1.08 vs 2.27+/-0.43 for kidney, 2.23+/-0.23 vs 0.17+/-0.02 for heart, 13.42+/-1.10 vs 5.72+/-1.01 for brain, respectively, P<0.05). However, no evident difference in the amount of carbonyl protein in liver was detected between SHR and WKY rats. (3) Total antioxidant capacities of kidney, liver, heart and brain were markedly lower in SHR than that in WKY rats (P<0.05). Thus, the present data reveal a higher inflammatory status in the important tissues in SHR and indicate that inflammation might play a potential role in pathogenesis of hypertension and secondary organ complications.
Animals ; Brain ; metabolism ; pathology ; Cytokines ; genetics ; metabolism ; Hypertension ; pathology ; Inflammation ; pathology ; Interleukin-1beta ; genetics ; metabolism ; Kidney ; metabolism ; pathology ; Male ; Myocardium ; metabolism ; pathology ; Oxidative Stress ; immunology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

In order to observe the proliferative and apoptotic situation of megakaryocytes in patients with myelodysplastic syndromes (MDS). CD41 immunoenzyme labeling (alkaline phosphatase anti-alkaline phosphatase APAAP)/DNA in situ end labelling (ISEL) double stained techniques was used onto plastic cold embedded bone marrow sections in 29 MDS patients to analyse the proliferative and apoptostic characterization of megakaryocytic line with 14 cases of iron deficient diseases (IDA) as control. The results showed that the mean CD41 positive cell number in MDS group was (26.23 +/- 8.18) /mm(2) with a count of (15.64 +/- 7.11) /mm(2) in control group (p < 0.05). The small-micro megakaryocytes in MDS is much higher than that in IDA group (P<0.01). There was a positive co-relation between total megakaryocytes and small-micro megakaryocytes count in MDS (r = 0.702, p<0.01). Some megakaryocytes distributed abnormally around trabecula and formed small or large clusters. Apoptotic megakaryocytes in MDS occupied 4.40% and 9.32% of all CD14 positive cells and all apoptotic cells respectively (p > 0.5 comparing with control). Apoptosis in megakargocytic line occurred only in small-micro megakaryocytes and showed positive co-relation to the number of micro-megakaryocytes. Some apoptotic cell with morphologic characters of megakaryocytes expressed no CD41. It is concluded that overproliferation of megakaryocytes exists in MDS. Apoptosis occurring in micro-megakaryocytes may be a kind of physiological response to abnormal megakaryopoicsis in MDS. No obvious increased apoptosis of megakaryocytes in MDS was found perhaps due to lack of surface antigens CD41 in some later stages of apoptotic cell.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Apoptosis ; Cell Division ; Child ; Female ; Humans ; Male ; Megakaryocytes ; pathology ; Middle Aged ; Myelodysplastic Syndromes ; pathology