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 .

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

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

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.

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.

To investigate the relationship between cytoskeleton and hyposmotic membrane stretch-induced increase in muscarinic current, the role of actin microfilament in hyposmotic membrane stretch-induced increase in muscarinic current was studied with the whole-cell patch clamp technique in guinea-pig gastric myocytes. In this study, the muscarinic current was induced by carbachol (50 micromol/L) or GTPgammaS (0.5 mmol/L). The results showed that hyposmotic superfusate (202 mOsmol/L) increased carbachol-induced current (I(CCh)) by 145+/-27% and increased GTPgammaS-induced current by 183+/-30%; but in the presence of cytochalasin-B (Cyt-B, 20 micromol/L), an actin cytoskeleton disruptor, hyposmotic membrane stretch increased I(CCh) by 70+/-6%. However, hyposmotic membrane stretch induced increase in I(CCh) was potentiated to 545+/-81% by phalloidin (20 micromol/L), an actin microfilament stabilizer. The results demonstrated that hyposmotic membrane stretch increased the muscarinic currents induced by carbachol or GTPgammaS and that the actin microfilament is involved in the process in guinea-pig gastric myocytes.
Actin Cytoskeleton ; physiology ; Animals ; Carbachol ; pharmacology ; Female ; Guinea Pigs ; Male ; Membrane Potentials ; drug effects ; Myocytes, Smooth Muscle ; physiology ; Osmotic Pressure ; Patch-Clamp Techniques ; Pyloric Antrum ; cytology ; Receptors, Muscarinic ; physiology

OBJECTIVESTo study the correlation between positive rates of RNA in clinical confirmed severe acute respiratory syndrome (SARS) patients and its appearance in relation to the development of the disease in order to provide scientific basis for early diagnosis, effective prevention and treatment of the disease.

METHODSOne-step reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the SARS RNA in the clinical specimens from different courses of the disease. The representative amplicons were then sequenced. Chi-square for trend test was performed to study the correlation between positive rates of RT-PCR and at different periods after the onset of the disease.

RESULTSThe fragments amplified from the sputum specimens of SARS patients were shown to share 100% homology with the published SARS-associated coronavirus. Of the different clinical specimens, positive rate in the stools appeared to be the highest (21.55%). Chi-square for trend test revealed that the positive rates of stools and sputa of SARS patients decreased with the development of the disease (chi(2) for trend = 12.55 and 16.408, P = 0.0004 and P = 0.000 05 respectively).

CONCLUSIONOne-step RT-PCR proved to be an effective method for the detection of SARS-associated coronavirus from clinical specimens. Data as indicated that the positive rates of SARS coronavirus were decreasing in SARS patients along with the disease progression.

Adolescent ; Adult ; Aged ; Chi-Square Distribution ; China ; Disease Progression ; Feces ; virology ; Female ; Humans ; Male ; Middle Aged ; Mucus ; virology ; RNA, Viral ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; Severe Acute Respiratory Syndrome ; pathology ; virology