OBJECTIVETo investigate the role and significance of cardiac mast cells and Toll-like receptor 4 (TLR4) in the development and progression of viral myocarditis (VMC).

METHODSForty-eight Balb/c mice were randomly divided into a control group (n=24) and a model group (n=24). Coxsackievirus B3 was intraperitoneally injected into the model group mice to establish a VMC model. In each group, cardiac tissues were collected from 8 mice at 7, 14 and 28 days after the model was established. The cardiac tissues were stained with hematoxylin and eosin as well as Masson trichrome to observe pathological changes in cardiac tissues. The number and degranulation of cardiac mast cells at each time point were measured and evaluated by toluidine blue staining and transmission electron microscopy. The mRNA and protein expression of TLR4 in cardiac tissues was measured by RT-PCR and immunohistochemistry. In the model group, the correlation between number of cardiac mast cells and mRNA expression of TLR4 at all time points was analyzed.

RESULTSThe model group had significantly higher pathological scores of cardiac tissues than the control group at all time points (P<0.05). The myocardial collagen volume fraction in the model group at 28 days was significantly higher than in the control group at all time points and higher than in the model group at 7 and 14 days (P<0.05). At each time point, the model group had a significantly increased number of mast cells (P<0.05), and significantly increased mRNA and protein expression of TLR4 (P<0.05) compared with the control group. In the model group, the number of cardiac mast cells was positively correlated with the mRNA expression of TLR4 at all time points (R²=0.877, P<0.05).

CONCLUSIONSMice with VMC have significantly increased numbers of cardiac mast cells and expression of TLR4 compared with control mice at all time points, suggesting that mast cells and TLR4 may play important roles in the inflammatory response and fibrosis of VMC.

Animals ; Coxsackievirus Infections ; immunology ; Enterovirus B, Human ; Female ; Mast Cells ; physiology ; Mice ; Mice, Inbred BALB C ; Myocarditis ; immunology ; Myocytes, Cardiac ; pathology ; Toll-Like Receptor 4 ; analysis ; genetics ; physiology

Many governments have actively encouraged the development of drug for rare diseases.In recent years, a series of policies has been pub-lished to encourage the research of rare diseases in china , and there are a large number of orphan drugs to apply for clinical trials or approval appli-cations now.This article summarizes the basic questions of clinical re-view in application of drugs for rare diseases , which will provide some references for the research of drugs for rare diseases .

ObjectiveTo evaluate the neurobiological characteristics of human histio-amniotic mesenchymal (hAMCs) and effect of hAMCs transplantation into the brain to treat Parkinson's disease(PD) modle mice.MethodsThe expressions of mesenchymal stem cells, neural stem cells, dopaminergic neurons and markers related to neurogenesis such as Vimentin, STRO-1, nestin, CD133, β-tubulin, TH, DAT, Ngn2 and mash-1 in hAMCs were evaluated through immunocytochemical stain; and the mRNA transcriptions of neural stem cell markers, Vimentin and nestin in hAMCs were detected by RT-PCR. The PD model was induced by MPTP(i.p.) in C57BL/6 mice transplanted with hAMCs into the right striatum. The therapeutical effect of hAMCs on PD mice was evaluated by spontaneous movement, rotating bar test and the immunohistochemistry of anti-human chondrosome and TH antibodies in striatum.ResultshAMCs induced by nerve cells culture medium, expressed mesenchymal stem cells, neural stem cells, dopaminergic neurons and other specific markers related to neurogenesis mentioned above. The frequency of spontaneous movement in PD mice was significantly increased(P<0-05), and the time of rotating bar was obviously prolonged(P<0-05) after transplantation with hAMCs.ConclusionhAMCs possess the characteristics of nerve cells after cultured in vitro and can significantly recover the damage of motor function induced by MPTP after transplantation into striatum in PD model mice.

OBJECTIVETo assess the effects of treatment of Amanita mushroom poisoning with Glossy anoderma Decoction (, GGD).

METHODSTwelve patients with acute Amanita mushroom poisoning received conventional treatment (penicillin and reduced glutathione) combined with oral administration of GGD (treated group), which was prepared out of 200 g Glossy ganoderma decocted in water to 600 mL, and 200 ml was given once, three times a day for 7 successive days; while conventional treatment alone was given to the other 11 patients assigned to the control group. The therapeutic efficacy and changes in serum levels of total bilirubin (TBil), bile acids (BA), alanine transaminase (ALT), and aspartate transaminase (AST) activities in the two groups were compared.

RESULTSThe cured-markedly effective rate in the treated group was more significant than that in the control group (P<0.01). Elevation in TBil, BA, ALT, and AST activities were observed in both groups 3 days after poisoning, which progressively increased thereafter in the control group. In the treated group, they reached their peak on the 3rd day and then declined gradually. The differences between pre-treatment and post-treatment in both groups were obviously significant (P<0.01), so were the differences between the two groups at corresponding time points (P<0.01).

CONCLUSIONGGD shows excellent clinical efficacy in the treatment of acute Amanita mushroom poisoning and can reduce mortality significantly.

Acute Disease ; Adolescent ; Adult ; Amanita ; Bile Acids and Salts ; blood ; Child ; Female ; Ganoderma ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Mushroom Poisoning ; blood ; drug therapy ; mortality

OBJECTIVEA total of 27 samples belonging to 5 cultivars of Fructus Aurantii (Citrus aurantium), i. e. cv. Xiucheng, cv. Xiangcheng, cv. Lecheng, cv. Jizicheng, and cv. Youzicheng, collected at Changfu and Huanggang, Zhangshu City, Jiangxi Province, were assayed to reveal the genetic relationship among the cultivars and the accordance between morphological and molecular markers.

METHODCultivar identification was based on morphology and cultivar relationship was based on Inter-simple sequence repeats (ISSR).

RESULTTwenty out of 40 ISSR primers screened generated 392 loci across all 27 samples with 315 informative loci. The UPGMA dendrogram showed that samples within cv. Xiucheng and cv. Xiangcheng from Changfu were closely related. However, samples of cv. Lecheng, cv. Jizicheng and cv. Youzicheng from Huanggang, or cv. Xiucheng and cv. Xiangcheng from both Changfu and Huanggang did not exhibited close relationships within each cultivars.

CONCLUSIONBased on morphology the same cultivar grown in different plantations, or even within a single plantation sometimes do not show close genetic relationship, indicating diverse origin of the cultivars. Synonyms or homonyms are believed to common phenomenon in Fructus Aurantii production. To solve the problem ISSR markers can serve a kind of molecular markers which are preferable to partition genetic variations within and between cultivars and to establish genetic relationships among them.

Citrus ; anatomy & histology ; classification ; genetics ; DNA Primers ; DNA, Plant ; genetics ; Fruit ; anatomy & histology ; genetics ; Genetic Markers ; Genetic Variation ; Phylogeny ; Plants, Medicinal ; anatomy & histology ; classification ; genetics ; Repetitive Sequences, Nucleic Acid

OBJECTIVETo study the chemical constituents in seeds of Taxus mairei.

METHODPreparative HPLC, TLC and spectroscopic analyses were used to isolate and elucidate the chemical constituets in the plant.

RESULTSeven taxane diterpenoids were isolated from the seeds of T. mairei and identified as taxinine A(1), 9-deacetyltaxinine(2), 9-deacetyltaxinine E(3), 2-deacetyltaxinine(4), taxezopidine G(5), 2-deacetoxytaxinine J(6), 2-deacetoxytaxuspine C(7).

CONCLUSIONExcept compounds 5,6, all the compounds were obtained from seeds of this plant for the first time.

Plants, Medicinal ; chemistry ; Seeds ; chemistry ; Taxoids ; chemistry ; isolation & purification ; Taxus ; chemistry

Objectives To investigate the technique and efficiency of stapled anastomoses in the ultra-low anterior dissection of rectal cancer.Methods From Oct 2000 to Oct 2006，the clinical data of 180 cases with rectal cancer undergone ultra-low anterior dissection were reviewed.Results During anastomosis，the distant closure end was tore apart in 3 cases(1.7％)，failure of anastomosis was encountered in 2 cases(1％)，and the anastomosis ring was incomplete in 7 cases.Postoperation anastomotic leakage happened in 6 cases(3.2％，6/180)，anastomotic stenosis occurred in 32 cases (17.7％，32/180).The average defecation was(4.3±5.2)times per day during hospitalization，and (2.5±3.1)times per day 2 months later，and normalized 3 months after surgery.Conclusion While ensuring the oncological excision and efficiency of anastomosis，the application of stapled anastomoses in the ultra-low anterior dissection of rectal cancer facilitates the preservation of anal sphincter function.

Objectives To investigate the technique and efficiency of stapled anastomoses in the ultra-low anterior dissection of rectal cancer.Methods From Oct 2000 to Oct 2006，the clinical data of 180 cases with rectal cancer undergone ultra-low anterior dissection were reviewed.Results During anastomosis，the distant closure end was tore apart in 3 cases(1.7％)，failure of anastomosis was encountered in 2 cases(1％)，and the anastomosis ring was incomplete in 7 cases.Postoperation anastomotic leakage happened in 6 cases(3.2％，6/180)，anastomotic stenosis occurred in 32 cases (17.7％，32/180).The average defecation was(4.3±5.2)times per day during hospitalization，and (2.5±3.1)times per day 2 months later，and normalized 3 months after surgery.Conclusion While ensuring the oncological excision and efficiency of anastomosis，the application of stapled anastomoses in the ultra-low anterior dissection of rectal cancer facilitates the preservation of anal sphincter function.

This study was to investigate the changes of autonomic nerve function and hemodynamics in patients with vasovagal syncope (VVS) during head-up tilt-table testing (HUT).HUT was performed in 68 patients with unexplained syncope and 18 healthy subjects served as control group.According to whether bradycardia,hypotension or both took place during the onset of syncope,the patients were divided during the test into three subgroups:vasodepressor syncope (VD),cardioinhibitory syncope (CI) and mixed syncope (MX) subgroups.Heart rate,blood pressure,heart rate variability (HRV),and deceleration capacity (DC) were continuously analyzed during HUT.For all the subjects with positive responses,the normalized low frequency (LFn) and the LF/HF ratio markedly decreased whereas normalized high frequency (HFn) increased when syncope occurred.Syncopal period also caused more significant increase in the power of the DC in positive groups.These changes were more exaggerated compared to controls.All the patients were indicative of a sympathetic surge in the presence of withdrawal vagal activity before syncope and a sympathetic inhibition with a vagal predominance at the syncopal stage by the frequency-domain analysis of HRV.With the measurements ofDC,a decreased vagal tone before syncope stage and a vagal activation at the syncopal stage were observed.The vagal tone was higher in subjects showing cardioinhibitory responses at the syncopal stage.DC may provide an alternative method to understand the autonomic profile of VVS patients.

Objective To understand the biochemical characteristics and 16S rDNA genetic sequence evolution of strains isolated from diarrhea specimens so as to provide basis for classification and identification of Klebsiella pneumoniae. Methods Specimens were cultured using MacConkey and SS medium. All isolates were identified as K. pneumoniae by automated biochemical tests. DNA was extracted, 1500 bp fragments of the 16S rDNA gene were by amplified PCR and sequenced with K. pneumoniae 16S rDNA primer, after being cut. Fragments of 1000 bp overlapping sequences were analyzed by Blastn to confirm the identity of the isolates. A phylogenetic tree was constructed by PHYLIP process to analyze the 16S rDNA sequence of the isolated strain with other relative bacteria species in the GenBank databases. Results Among 113 specimens of infectious diarrhea, 25 K. pneumoniae strains were identified by biochemical tests, of which 21 subsp, pneumoniae and 4 subsp, ozaenae, no subsp, of rhinoseleroma were isolated. Strains of subsp, pneumoniae were found having nature of resistance. All isolates were resistant to penicillin G and susceptible to polymyxin with some strains were resistant to Nitrofurantoin, Cephalothin, Kanamycin, Tobramycin. After searching in GenBank of 16S rDNA, strains biochemical identified as subsp, ozaenae shared high similarity with Salmonella strains and other intestinal bacteria. 16S rDNA phylogenetie analysis could be used to confirm subsp, pneumoniae, but could not separate other subspecies of K. pneumoniae completely. Conclusion 16S rDNA phylogenetic analysis useful in identifying and classifying K. pneumoniae.