Objective To investigate the relationship between expression of surfactant protein B (SP-B) gene product and neonatal respiratory distress syndrome (NRDS) in Han ethnic group.Methods Unrelated 20 cases with NRDS of Han ethnic group were selected as NRDS group while unrelated 20 cases of Han ethnic group with other diseases were selected as a control. The cases in the control group had congenital heart disease or bronchopulmonary dysplasia or persistent pulmonary artery hypertension. Blood sample was taken from each case. Lung tissues were taken from the patients in half an hour after their death in the two groups. Expression of SP-B in the lung tissues was determined with immunohistochemical technique. Genetic deficiency variant of SP-B intron Ⅳ was screened with polymerase chain reaction (PCR).Results Two cases at gestational age of 26 weeks, one at 34 weeks and two at 42 weeks in the NRDS groups had lower expression level of SP-B in the lung tissues than those at the same age in the the control group. Expression of SP-B in the lung tissues of the control group increased with gestational age, but no such phenomenon was found in NRDS group. Further two cases at gestational age 42 weeks of NRDS group had genetic deficiency variant of SP-B intron Ⅳ with gene analysis of five cases who had lower expression of SP-B. Clinical data suggest that patients at 42 weeks of gestational age had severe illness.Conclusion Decrease of SP-B expression may be involved in occurrence of NRDS, genetic deficiency variant of SP-B intron Ⅳ exists in NRDS cases of Han ethnic group of China.

Objective To determine if there is any difference in neuronal and glial(astrocytic and microglial)activation in the spinal cord in three rat models of neuropathic pain.Methods Twenty-four SD rats weighing 150-200 g were randomly divided into 4 groups(n=6 each):Ⅰ control group;Ⅱ chronic constrictive injury group(CCI);Ⅲ spinal nerve ligation group(SNL)and Ⅳ spared nerve injury(SNI).No operation was performed in control group.In CCI group left sciatic nerve was exposed and loosely ligated with catgut.In SNL group the L_5 spinal nerve was exposed and ligated with silk suture and cut.In SNI group tibial nerve and common fibular nerve were ligated and cut.Pain threshold was measured using plantar tactile stimulator(Ugo,Basile Co. Italy)every other day from 3 days before until 15 days after operation.50% paw withdrawal threshold was measured using up-and-down sequential mechanical stimulation of different intensity(0.45,0.70,1.20,2.00, 3.63,5.50,8.50,15.10 g)applied to the plantar surface of the injured paw.On the 15~(th) day after operation after pain threshold was measured the animals were anesthetized with intraperitoneal 10% chloral hydrate 400 mg? kg~(-1).The L_(5,6) segment of the spinal cord was isolated.Neuronal,astrocytic and microglial activation was determined by immuno-histochemistry with antibodies of c-Fos(a proto-oncogene protein),GFAP(an astrocyte marker)and OX-42(a microglial marker).Results The 50% paw withdrawal threshold reached the lowest level on the 7~(th) day after operation.The lowest level was maintained until the 15~(th) day after operation in group CCI,SNL and SNI.The 50 % paw withdrawal threshold was(14.1+1.5)g in control group,(2.5+0.5)g in CCI group, (1.5?0.6)g in group SNL and(0.8?0.4)g in group SNI.The number of c-Fos positive neurons in laminae Ⅳ-Ⅵ of dorsal horn was significantly greater in group CCI,SNL and SNI than in control group,but there was no significant difference among the 3 peripheral nerve injury groups.The activation of astrocytes and microglias in laminae Ⅰ-Ⅳ of dorsal horn was significantly increased in group CCI,SNL and SNI than in control group but there was no significant difference among the 3 peripheral nerve injury groups.Condusion There is no significant difference in activation of neurons and astrocytes and microglias in the ipsilateral dorsal horn among the 3 pain models.

Anesthetics, Intravenous ; pharmacology ; Animals ; Aorta, Thoracic ; drug effects ; physiology ; Endothelium, Vascular ; physiology ; Female ; In Vitro Techniques ; Male ; Norepinephrine ; pharmacology ; Potassium Channels ; physiology ; Propofol ; pharmacology ; Rabbits ; Vasodilation ; drug effects

OBJECTIVETo explore the application of caffeine-halothane contracture test (CHCT) in the confirmation of malignant hyperthermia (MH).

METHODSOne patient who underwent radical gastrectomy presented with clinical manifestations of MH during routine intravenous-inhalation anesthesia process. Isoflurane inhalation and the operation were ceased immediately and emergency management approaches such as physical cooling therapy were taken. Meanwhile, the levels of serum creatine kinase (CK), serum myoglobin, and urinary myoglobin were examined and rectus abdominis was taken and then CHCT was performed to confirm the clinical diagnosis. Total genome was extracted from the patient and then exons 2-18, 39-46, and 90-104 of ryanodine receptor 1 (RYR1) gene were screened to detect mutations using DNA sequencing technique.

RESULTSThe patient was diagnosed as MH episode by clinical characteristics and postoperatively continuous elevation of the levels of CK, serum myoglobin, and urinary myoglobin (30 times higher than normal level). Despite halothane test was negative, the diagnosis of MH was verified by the positive result of caffeine test. DNA sequencing of RYR1 gene of the patient revealed c. 6724C > T (p. T 2 206M).

CONCLUSIONCHCT can be used to confirm the diagnosis of MH.

Anesthetics, Inhalation ; therapeutic use ; Caffeine ; Creatine Kinase ; blood ; Enzyme-Linked Immunosorbent Assay ; Halothane ; Humans ; Isoflurane ; therapeutic use ; Malignant Hyperthermia ; blood ; diagnosis ; genetics ; Muscle, Skeletal ; drug effects ; physiopathology ; Myoglobin ; blood ; Ryanodine Receptor Calcium Release Channel ; genetics

The medicinal fungi, which are of great importance in traditional medicine, are facing the problems of wild resources scarcity and low concentration of bioactive compounds. Velvet family and LaeA global regulator play a vital role in secondary metabolism and developmental programs, which are found in a wide variety of fungi ranging from Chytridiomycota to Basidiomycota. This review elaborates the structures and functions between Velvet family and LaeA protein. The Velvet family which shares the Velvet protein domain, including VeA (Velvet), VelB (Velvet like B), VosA (viability of spores A) and VelC (Velvet like C), acts on the regulation function is secondary metabolism and developmental programs such as asexual and sexual development. Furthermore, the function is affected by environmental factors such as light and temperature. LaeA protein which owns S-adenosylmethionine-dependent methyltransferase domain, coordinately regulates development and secondary metabolism by regulating and modifying the Velvet proteins. The regulation of LaeA is mediated by light receptor proteins. Therefore, clarifying the mechanism of Velvet and LaeA proteins in medicinal fungi will pave the way for nurturing medicinal fungi and improving production of bioactive compounds.
Fungal Proteins ; metabolism ; Fungi ; chemistry ; Gene Expression Regulation, Fungal ; Genes, Regulator ; Protein Structure, Tertiary ; Secondary Metabolism ; Structure-Activity Relationship

OBJECTIVETo observe the effect of adenovirus-mediated overexpression of AMP-activated protein kinase (AMPK) on apoptosis of hepatic stellate cell line LX2.

METHODSAfter adenovirus infection of the LX2 cells, the exogenous gene expression was detected by Western blotting, and cell apoptosis evaluated by flow cytometry with PI staining. Agarose gel electrophoresis was performed to detect the DNA ladder, and the expressions of caspase-3, Bcl-2 and Bax are detected by Western blotting.

RESULTWith AMPK overexpression, apoptotic peak and DNA ladder of the infected cells appeared, and pro-caspase-3 was activated to transform into caspase-3 accompanied by up-regulated Bax expression, whereas Bcl-2 expression was hardly detectable by Western blotting.

CONCLUSIONOverexpression of AMPK mediated by the adenovirus can induce LX2 cell apoptosis, possible as a result of up-regulated Bax expression with low Bcl-2 expression.

AMP-Activated Protein Kinases ; genetics ; metabolism ; Adenoviridae ; genetics ; Apoptosis ; Blotting, Western ; Caspase 3 ; metabolism ; Cell Line ; Gene Expression Regulation, Enzymologic ; Genetic Vectors ; genetics ; Hepatic Stellate Cells ; cytology ; metabolism ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Transduction, Genetic ; bcl-2-Associated X Protein ; metabolism

BACKGROUNDMalignant hyperthermia (MH), manifesting as MH crisis during and/or after general anesthesia, is a potentially fatal disorder in response to volatile anesthetics and depolarizing muscle relaxants. Though typical features of MH episode can provide clues for clinical diagnosis, MH susceptibility is confirmed by in vitro caffeine-halothane contracture test (CHCT) in western countries. It is traditionally thought that MH has less incidence and fewer typical characteristics in Chinese population than their western counterparts because of the different genetic background. In this study, we investigated the clinical features of MH in Chinese cases and applied the clinical grading scale and CHCT for diagnosis of MH.

METHODSA cluster of three patients with MH, from January 2005 to December 2007, were included in the study. Common clinical presentations and the results of some lab examinations were reported in detail. The method of the clinical grading scale of diagnosis of MH was applied to estimate the qualitative likelihood of MH and predict MH susceptibility. Muscle fibers of femoral quadriceps of the patients were collected and CHCT was performed to confirm the diagnosis of MH.

RESULTSThe clinical grading scales of diagnosis of the disease for these cases were all ranked grade D6, suggesting almost diagnosed ones. And the results of caffeine test were positive correspondingly, indicating that the patients should be diagnosed as MH susceptibility (MHS) according to diagnostic criteria of the North America MH group, which were already confirmed by clinical presentations and biochemical results.

CONCLUSIONSThese Chinese cases manifest as MH crisis. The clinical grading scale of diagnosis of MH may provide clues for clinical diagnosis. CHCT can also be used in confirming diagnosis of MH in Chinese cases though they have different genetic background from their western counterparts.

Adolescent ; Adult ; Caffeine ; Child ; China ; Female ; Halothane ; Humans ; In Vitro Techniques ; Male ; Malignant Hyperthermia ; diagnosis ; Muscle Contraction ; drug effects ; Young Adult

A series of adefovir mono-L-amino acid esters, mono non-steroidal anti-inflammatory drugs carboxylic ester prodrugs with more potent anti-HBV activity and lower nephrotoxicity were designed and synthesized. Adefovir bis (L-amino acid) ester was used as lead compound, according to pathological and pharmacological findings that non-steroidal anti-inflammatory drugs can effectively inhibit the organic anion transporter 1 (hOAT1)-mediated adefovir phosphonic acid pairs of anion transport across tubular basement membrane thereby reducing the nephrotoxicity of adefovir. Flatten design principle was used to introducing non-steroidal anti-inflammatory drugs structural fragments to design and synthesize target adefovir mixture ester prodrugs. HepG2 2.2.15 cell line was used as in vitro anti-HBV activity evaluation model. Five compounds exhibited antiviral activity, and compound 18 showed the most potent anti-HBV activity and relatively high selective index (EC50 3.92 micromol L(-1), SI 9.97). HK-2 cell line was used as in vitro model to evaluate nephrotoxicity. Results suggested the target compounds have lower cytotoxicity than the positive control. Moreover, by analyzing the primary structure and activity relationship of these compounds, it could suggest that mono-L-amino acid ester, mono non-steroidal anti-inflammatory drugs carboxylic ester prodrugs strategy has significant potential in the acyclic nucleoside phosphonates prodrug design.
Adenine ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Amino Acids ; chemical synthesis ; chemistry ; pharmacology ; Anti-Inflammatory Agents, Non-Steroidal ; chemical synthesis ; chemistry ; pharmacology ; Antiviral Agents ; chemical synthesis ; chemistry ; pharmacology ; Carboxylic Acids ; chemistry ; pharmacology ; Cell Survival ; drug effects ; Drug Design ; Hep G2 Cells ; drug effects ; Humans ; Kidney Tubules, Proximal ; cytology ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Molecular Structure ; Organophosphonates ; chemical synthesis ; chemistry ; pharmacology ; Prodrugs ; chemical synthesis ; chemistry ; pharmacology

Transcription factor is one of the key factors in the regulation of gene expression at the transcriptional level. It plays an important role in plant growth, active components biosynthesis and response to environmental change. This paper summarized the structure and classification of bHLH transcription factors and elaborated the research progress of bHLH transcription factors which regulate the active components in plants, such as flavonoids, alkaloids, and terpenoids. In addition, the possibility of increasing the concentration of active substances by bHLH in medicinal plants was assessed. The paper emphasized great significance of model plants and multidisciplinary research fields including modern genomics, transcriptomics, metabolomics and bioinformatics, providing the contribution to improve the discovery and function characterization of bHLH transcription factors. Accelerating the research in the mechanism of bHLH transcription factors on the regulation of active components biosynthesis will promote the development of breeding and variety improvement of Chinese medicinal materials, also ease the pressure of resources exhaustion of traditional Chinese medicine home and abroad.
Alkaloids ; biosynthesis ; Basic Helix-Loop-Helix Transcription Factors ; chemistry ; classification ; genetics ; metabolism ; Flavonoids ; biosynthesis ; Plants, Medicinal ; genetics ; metabolism ; Terpenes ; metabolism