Abstract] Objective To investigate the in vivo therapeutic effects of an extract of herb medi-cines, YiGanQingDuKeLi, in combination with adefovir dipivoxil (ADV) on the rebound of duck hepatitis B virus ( DHBV) multiplication after withdrawal of ADV treatment.Methods Peking ducks were infected with DHBV positive serum samples for 7 days and then screened by SYBR Green real-time PCR.The ducks positive for DHBV were randomly divided into five groups including the model control group, the ADV treat-ment group, the herb treatment group, the high-dose combination therapy group and the low-dose combina-tion therapy group.The ducks in the ADV treatment and the herb treatment groups were respectively treated with distilled water and YiGanQingDuKeLi (1.2 g/ml) for 14 days after the treatment of ADV (0.25 mg/ml) for 21 days.The ducks in the high-dose group were treated with YiGanQingDuKeLi (1.2 g/ml) for 14 days after the combined treatment with high-dose YiGanQingDuKeLi (1.2 g/ml) and ADV (0.25 mg/ml) for 21 days.The ducks in the low-dose group were treated with YiGanQingDuKeLi (0.6 g/ml) for 14 days after the combined treatment with YiGanQingDuKeLi (0.6 g/ml) and ADV (0.125 mg/ml) for 21 days.Blood samples were collected from each duck via leg vein after 0, 7, 14 and 21 days of drug adminis-tration and after 7 and 14 days of drug withdrawal.The levels of DHBV-DNA, alanine aminotransferase ( ALT) and aspartate aminotransferase ( AST) in blood serum samples were detected.Results Compared with the model group, the levels of DHBV-DNA, ALT and AST in ducks from the herb treatment group and combined treatment groups were decreased before the discontinuation of ADV treatment ( P<0.05 or P<0.01).Moreover, the titers of DHBV-DNA in ducks treated with high doses of drugs were much lower than those from ADV treatment group.The levels of DHBV-DNA, ALT and AST in ducks treated with herb medi-cine and high doses of drugs remained at relatively low levels after the cessation of ADV treatment, but re-bounded significantly in ducks with ADV treatment.The levels of DHBV-DNA and ALT rebounded slightly in ducks treated with low doses of drugs as compared with those of ADV treatment group ( P<0.01 or P<0.05).Conclusion The treatment of YiGanQingDuKeLi in combination with ADV could inhibit not only the in vivo replication of DHBV, but also the rebound of DHBV multiplication after ADV withdrawal.

Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma were widely used in strengthening spleen under different disease conditions, and were easily and often misused each other. Therefore, DNA barcode was used to distinguish Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma from their adulterants to ensure the safe use. The sequence lengths of ITS2 of Atractylodes macrocephala, Atractylodis Rhizoma (A. lancea, A. japonica and A. coreana) were both 229 bp. Among the ITS2 sequences of A. macrocephala, only one G/C transversion was detected at site 98, and the average GC content was 69.42%. No variable site was detected in the ITS2 sequences of A. lancea. The maximum K2P intraspecific genetic distances of both A. japonica and A. coreana were 0.013. The maximum K2P intraspecific genetic distances of A. macrocephala, A. lancea, A. japonica and A. coreana were less than the minimum interspecific genetic distance of adulterants. The ITS2 sequences in each of these polytypic species were separated into pairs of divergent clusters in the NJ tree. DNA barcoding could be used as a fast and accurate identification method to distinguish Atractylodis Macrocephalae Rhizoma, Atractylodis Rhizoma, from their adulterants to ensure its safe use.
Atractylodes ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics

OBJECTIVETo establish discriminant functions of diarrhea-predominant irritable bowel syndrome (IBS-D) by studying it from quantitative diagnosis angle, hoping to reduce interference of subjective factors in diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.

METHODSA Chinese medical clinical epidemiological survey was carried out in 439 IBS-D patients using Clinical Information Collection Table of IBS. Initial syndromes were obtained by cluster analysis. They were analyzed using step-by-step discrimination by taking information of four Chinese medical diagnostic methods and serum brain-gut peptides (BGP) as variables.

RESULTSClustering results were Gan stagnation Pi deficiency syndrome (GSPDS), Pi-Wei weakness syndrome (PWWS), Gan stagnation qi stasis syndrome (GSQSS), Pi-Shen yang deficiency syndrome (PSYDS), Pi-Wei damp-heat syndrome (PWDHS), cold-damp disturbing Pi syndrome (CDDPS). Of them, GSPDS was mostly often seen with effective percentage of 34. 2%, while CDDPS was the least often seen with effective percentage of 5.5%. A total of 5 discriminant functions for GSPDS, PWWS, GSQSS, PSYDS, and PWDHS were obtained by step-by-step dis- crimination method. The retrospective misjudgment rate was 4.1% (16/390), while the cross-validation misjudgment rate was 15.4% (60/390).

CONCLUSIONThe establishment of discriminant functions is of value in objectively diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.

Alarmins ; Brain ; Cluster Analysis ; Diarrhea ; classification ; diagnosis ; Hot Temperature ; Humans ; Irritable Bowel Syndrome ; classification ; diagnosis ; Medicine, Chinese Traditional ; Qi ; Retrospective Studies ; Surveys and Questionnaires ; Yang Deficiency

OBJECTIVETo observe the effect of Panax notoginseng saponins (PNS) on serum content of neuronal specific enolase (NSE) and function recovery in patients with cerebral hemorrhage (CH) for exploring the therapeutic action of PNS in treating CH.

METHODSFifty CH patients with their course of disease not more than 5 days were randomly assigned to two groups, 27 in the PNS group and 23 in the control group, all were treated with conventional treatment, while PNS was given additionally to the PNS group. The serum levels of NSE before and after treatment were determined by electrochemiluminescence, and the recovery of patients, including their neuro-function deficit and daily living activity, was assessed according to scoring by the National Institute of Health Stroke Scale (NIHSS) and Barthel Index (BI) respectively.

RESULTSBefore treatment, the difference between the two groups in serum NSE content and scores of NIHSS and BI was insignificant (P > 0.05). However, after 3 weeks of treatment, the level of NSE and score of NIHSS were significantly lower, while score of BI was significantly higher in the PNS group than those in the control group respectively (all P < 0.01). In the PNS group, the level of NSE showed a positive correlation with the score of NIHSS (r = 0.757, P < 0.05), and a negative correlation with the score of BI (r = - 0.803, P < 0.05).

CONCLUSIONPNS can effectively protect the neuron and promote functional rehabilitation in patients after CH.

Adult ; Aged ; Biomarkers ; blood ; Cerebral Hemorrhage ; blood ; drug therapy ; rehabilitation ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Panax notoginseng ; chemistry ; Phosphopyruvate Hydratase ; blood ; Phytotherapy ; Saponins ; therapeutic use ; Treatment Outcome

Objective To investigate the curative effect of brucellar spondylitis,so as to provide scientific proof for improving the curative level of the disease.Methods Epidemiological information was collected from 113 patients diagnosed as brucellar spondylitis,who were divided into 5 groups according to different drugs and drug combinations of doxycycline,gentamicin,sulfamethoxazole and rifampicin.Then the curative effect was investigated.Twenty-one patients who had greater paoas muscle abscess or Para vertebral abscess,intraspinal abscess of spinal canal,necrotic intervertebral disk and major osteolasia received the minimal invasive surgery and the focus removal surgery.Results The occurrence of the disease in female was much higher than that in male.Grazing and breeding beasts was the principal route of infection.Lumbars was mostly involved.they usually was infected in the adjacent 2 piece.L4 was the most common and seriuous one.The curative effect of doxycycline group was better than that without doxycycline(72.60% vs 35.00%,X2=15.14,P＜0.05).Doxycyeline+gentamicin+sulfamethoxazole was reeommended as the first choice.However,the curative effect did not increase despi~course of the treatment prolonged.The heMing rate and effective rate after 1 course was 52.21%(59/113)and 92.04%(104/113).that after 2 courses 58.41%(66/113)and 95.58%(108/113),that after 3 courses 59.29%(67/113)and 95.58%(108/113).The healing rate in different course did not present differences(P＞0.05).21 patients undergoing surgery were followed-up,12 patients were after 2 years and 9 patients were between 1-2 years.The healing rate was 95.24%(20/21),1 case was healed basically,the effective rate was 100%.None reoccurred.Conclusions There are characterized features in clinical epidemiology of the brucell spondylitis.Long term,adequate in dosage,combination and multi-approach use of antibiotics is the most reliable way to treat and prevent it from recurring.But fof the patients soitable for surgery.the minimal invasive surgery or the focus removal could shorten the course of therapy,decrease the complications and increase the cure rate.

Objective To explore the residual undifferentiated mouse embryonic stem cells (ESCs) in embryoid bodies. Methods Mouse R1 and Oct-4-GFP transgenic ESCs were firstly cultured in suspension to form embryoid bodies (EBs). Twenty days later, EBs were digested into single cells and then re-plated in standard ESC culture condition. The morphology of residual undifferentiated cells in EBs was observed, and surface makers and in vitro redifferentiation potency of residual cells were examined by flow cytometry and immunofluoreseent staining. The residual cells were expanded and subcutaneously injected into nude mice, and the specimens were harvested from the injection site for histological analysis 6 weeks after injection. Results There were residual undifferentiated ESCs in EBs differentiated for 20 days, which displayed clonal morphology and expressed undifferentiated cell markers of ESCs, including SSEA1, CD31, CD9 and Oct-4. The cells could be differentiated to form EBs again, and could be re-expanded from secondary EBs. The residual cells were able to form teratoma at the injection site, and mature endoderm, mesoderm and ectoderm tissues could be found in teratoma tissues. Conclusion There are residual undifferentiated ESCs after differentiation of ESCs into EBs. The residual ESCs can differentiate again in vitro and in vivo, and can residue again in the in vitro differentiation.

OBJECTIVETo investigate the genetic association between the polymorphism of cytosolic phospholipase A2 (cPLA2) family genes and schizophrenia in the North Han Chinese.

METHODSMethod of polymerase chain reaction-based ligase detection reaction (PCR-LDR) was applied to genotype 10 single nucleotide polymorphisms (SNPs) of cPLA2 family genes among 201 pedigrees consisting of fathers, mothers and affected offsprings with schizophrenia. Haplotype relative risk (HRR) test, transmission disequilibrium test (TDT), haplotype transmission analysis and multiple locus analysis were conducted to analyze the genotyping data.

RESULTSThe genotypic frequency of cPLA2 gene did not deviate from Hardy-Weinberg equilibrium in both case and control groups. HRR and TDT showed that the 10 SNPs were not associated with schizophrenia (P > 0.05). Analysis for haplotype transmission showed that no haplotype systems was associated with schizophrenia (P > 0.05). Results from COA and COG tests showed a disease association for the rs2162886-rs1668589, rs891014-rs1668589 and rs2307279-rs7542180 combinations (chi2 = 6.913, P = 0.032; chi2 = 8.393, P = 0.015; chi2 = 8.447, P = 0.038).

CONCLUSIONMany loci in the cPLA2 family genes were associated with schizophrenic.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; China ; epidemiology ; Female ; Gene Frequency ; genetics ; Genetic Predisposition to Disease ; genetics ; Genotype ; Haplotypes ; genetics ; Humans ; Male ; Phospholipases A2, Cytosolic ; genetics ; Polymorphism, Single Nucleotide ; genetics ; Schizophrenia ; epidemiology ; genetics ; Young Adult

HPLC-ELSD was applied to explore the absorption mechanism of pulchinenosides (B3, BD, B7, B10, B11) in rats. The experimental results showed that the absorption rate constant, Ka value (B3, BD) and Permeability coefficient, Peff value (B3, B7) displayed significant difference (P <0.05) in various intestinal segments, The Ka value and Peff value of PRS was different from each other with the highest absorption in duodenum (duodenum > jejunum > colon > ileum); The PRS displayed excessive satuation as the concentration increased over 0.05-2.5 g · L(-1). There were no obvious linear correlations between Peff values and concentrations in duodenum (0.6007 ≤ R2 ≤ 0.7727); Ka and Peff value declined when the PRS was perfused with P-glycoprotein promoter digoxin, on the other hand, inclined when perfused with P-glycoprotein inhibitor verapamil with significant difference among PRS B3, BD, B7, B11 (P <0.05). All the above results demonstrated that B3, BD, B7 were greatly influenced by absorption sites, duodenum was the main absorption site; PRS didn't entirely transported in a concentration dependent manner, and the transporter-protein involved the transportation, so the intestinal absorption of the five pulchinenosides was not entirely passive diffusion; and PRS might be the substrates of P-glycoprotein.
ATP-Binding Cassette, Sub-Family B, Member 1 ; physiology ; Animals ; Intestinal Absorption ; Male ; Oleanolic Acid ; pharmacokinetics ; Pulsatilla ; chemistry ; Rats ; Rats, Wistar ; Saponins ; pharmacokinetics

The paper reports the systematic study on felodipine and its impurities in tablets, to improve its quality standards for the control of the related substances. HPLC-DAD, UPLC-MS, IR and NMR methods were used for the isolation of felodipine and its impurities in tablets, their identification and the zebrafish animal model was used for the analysis of the toxic impurities. In felodipine material and its tablets, three impurities are isolated and identified. They are impurity 1 [dimethyl 4-(2, 3-dichlorophenyl)-2, 6-dimethyl-1, 4-dihydropyridine-3, 5-dicarboxylate], impurity 2 [ethyl methyl 4-(2, 3-dichlorophenyl)-2, 6-dimethylpyridine-3, 5-dicarboxylate] and impurity 3 [diethyl 4-(2, 3-dichlorophenyl)-2, 6-dimethyl-1, 4-dihydropyridine-3, 5-dicarboxylate], separately. The result of zebrafish animal model analysis showed that the teratogenic effects of four compounds were: impurity 3 > or = felodipine > impurity 1 > impurity 2, lethal effects were as follows: impurity 2 = impurity 3 > felodipine > or = impurity 1. This study confirmed the toxicity of three impurities in felodipine. According to the results, the paper suggested the amendments to the standard of the medicine and provided the support to the control of impurities in the manufacturing process.
Abnormalities, Drug-Induced ; Animals ; Antihypertensive Agents ; administration & dosage ; chemistry ; toxicity ; Calcium Channel Blockers ; administration & dosage ; chemistry ; toxicity ; Chromatography, High Pressure Liquid ; methods ; Drug Contamination ; Felodipine ; administration & dosage ; chemistry ; toxicity ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Pharmaceutical Preparations ; analysis ; chemistry ; Quality Control ; Spectrophotometry, Infrared ; Tablets ; Tandem Mass Spectrometry ; Zebrafish

Objective: To explore the effect of herb-partitioned moxibustion (HPM) on tight junctions (TJs) of intestinal epithelial cells in Crohn disease (CD) mediated by tumor necrosis factor-α (TNF-α)-nuclear factor kappa B (NF-κB)-myosin-light- chain kinase (MLCK) pathway. Methods: Forty-eight male Sprague-Dawley rats were randomly divided into a normal control (NC) group, a model control (MC) group, an HPM group and a mesalazine (MESA) group, with 12 rats in each group. Trinitrobenzene sulfonic acid (TNBS) was administered to establish CD models. When the model was confirmed a success, the HPM group rats were treated with HPM at Tianshu (ST 25) and Qihai (CV 6), while the MESA group rats were given MESA solution by lavage. When the intervention finished, the colonic epithelial tissues were separated, purified and cultured in each group to establish the intestinal epithelial barrier model in vitro, and TNF-α was added (100 ng/mL) in the culture medium and maintained for 24 h to establish an increased epithelial permeability model. Transepithelial electrical resistance (TEER) was used to examine the permeability of the barrier; Western blot was used to observe the expressions of the proteins related to TJs of intestinal epithelial cells mediated by TNF-α-NF-κB-MLCK pathway; immunofluorescence staining was used to observe the expressions and distributions of tight junction proteins in the intestinal epithelium. Results: After TNF-α induction, compared with the MC+TNF-α group, the TEER value increased significantly in the HPM+TNF-α and MESA+TNF-α groups (both P<0.001); the expressions of nuclear factor kappa B (NF-κB) p65, MLCK, myosin light chain (MLC), tumor necrosis factor receptor-associated factor 6 (TRAF6) and receptor interaction protein-1 (RIP1) decreased significantly (P<0.01 or P<0.05), and the expression of zinc finger protein A20 (A20) increased significantly (P<0.01); the expressions of occludin, claudin-1, zonula occludens protein 1 (ZO-1) and F-actin also increased significantly (all P<0.01). Compared with the MESA+TNF-α group, the expressions of MLC, occludin, claudin-1, ZO-1 and F-actin increased significantly in the HPM+TNF-α group (P<0.01 or P<0.05). Conclusion: HPM can protect or repair the damage of intestinal epithelial barrier in CD rats, which may be achieved through modulating the abnormal TJs in intestinal epithelium mediated by TNF-α-NF-κB-MLCK pathway.