OBJECTIVETo study the constituents with the pain-relieving activity from the stem rind of Daphne giraldii.

METHODThe partition of the ethanol extract and chromatographic separation of the fractions were carried out by the monitoring of anelgesic pharmacological activity. The structures of the isolated compounds were determined by MS and NMR.

RESULTFour compounds were isolated from the pain-relieving fraction. Three of them were identified as diterpenes, gniditrin (1), gnidicin (2) and daphnetoxin (3). Compound 4 was determined as Z-octadecyl caffeate.

CONCLUSIONCompounds 1, 2 and 4 were isolated from the plant for the first time.

Analgesics ; chemistry ; isolation & purification ; pharmacology ; Caffeic Acids ; chemistry ; isolation & purification ; pharmacology ; Daphne ; chemistry ; Diterpenes ; chemistry ; isolation & purification ; pharmacology ; Heterocyclic Compounds, 4 or More Rings ; chemistry ; isolation & purification ; pharmacology ; Molecular Structure ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry

Aim To study the effect of the total flavonoids of turpinia arguta seen (TFS) on immune function of rats with adjuvant arthritis. Methods Adjuvant arthritis (AA) was induced on d0 by intradermal injection of Complete Freund′s Adjuvant (FCA), containing 10 mg heat-inactive BCG in 1ml paraffin oil. On d 28 after immunization, AA rats were killed by cervical decollation. Immune cells (splenocytes and peritoneal macrophages) were obtained, then cultured in vitro with TFS. Some immune indexes were detected respectively such as the splenocytes proliferation and the productions of IL-1 and IL-2 were measured by the method of cell proliferation, the production of prostaglandin E2 was determined by radioimmunoassay. Results Compared with normal groups, the response of splenocytes to ConA and LPS was lowered, IL-2 syn-thesis was decreased, and IL-1 and PGE_2 released from PM? were elevated. TFS (10~ -8 ~10~ -4 mg?L~ -1 ) not only increased splenocytes proliferation, which induced by ConA and LPS, and IL-2 production of splenocytes, but also reduced the elevated productions of IL-1 and PGE_2 released from peritoned macrophate in AA rats in vitro. Conculsion TFS could adjust abnormal immune function in AA rats.

OBJECTIVEX-linked adrenoleukodystrophy (ALD) is a genetically determined disorder that involves the nervous system white matter, axons, adrenal cortex and testes. The typical clinical manifestations are progressive psychomotor regression, vision and/or auditory impairment and adrenal insufficiency. The clinical manifestation, biochemical change and genetic counseling work of X-linked ALD were analyzed.

METHODSThe clinical features of 29 cases with ALD were summarized and analyzed, including symptoms and signs, measurement of blood very long chain fatty acids (VLCFA), adrenal function, cranial magnetic resonance imaging (MRI) and pedigree investigation.

RESULTSAmong these 29 cases, the clinical phenotype could be classified into childhood cerebral (22 cases), adolescent cerebral (4 cases), adrenomyeloneuropathic (1 case), Addison's disease (1 case) and asymptomatic or presymptomatic (1 case) types. Nine of them had positive family history. Pedigree investigation was consistent with typical sex-linked recessive inheritance. There were 45 ALD patients in these 29 pedigrees. The neurological manifestations varied among members of the same family. Nine cases died during follow up. The causes of death were central respiratory failure or other complications of ALD and so on. Laboratory tests demonstrated abnormally high plasma levels of VLCFA in ALD patients; MRI demonstrated symmetric butterfly-like low T(1) and high T(2) signals in the parieto-occipital white matter. The impairment in the splenium of corpus callosum made the bilateral lesion region converge into one. It could progress anteriorly and injure the bilateral posterior limb of internal capsule and the temporal lobe, and could injure the brainstem inferiorly. Following intravenous injection of contrast material, thin stripe of lacelike enhancement could be observed.

CONCLUSIONSThe atypical initial symptom of ALD was seizures. The MRI showed abnormal signal in the cerebellar white matter. This disease can influence the normal development of children, this was more pronounced in the childhood cerebral ALD type. It tended to progress rapidly with dementia, vegetative state or death. Since antenatal diagnostic method is available now, emphasis should be made on the antenatal examination in order to make an early diagnosis and abort pregnancy if necessary.

Adolescent ; Adrenoleukodystrophy ; blood ; diagnosis ; therapy ; Child ; Child, Preschool ; China ; Fatty Acids ; blood ; Female ; Follow-Up Studies ; Humans ; Male ; Pedigree ; Treatment Outcome

OBJECTIVETo investigate clinicopathological and immunohistochemical features of 31 cases of oral teratoid cyst.

METHODSThirty-one cases of oral teratoid cyst according with Meyer's diagnosis criteria were retrospectively studied and their histochemistry and immunohistochemistry were performed from the files of Department of Pathology, School of Stomatology, Wuhan University between 1963 and 2003.

RESULTSTwenty-seven cases (87.10%) were children, and twenty-four cases were congenital. The ratio of male-to-female was 1:0.55, and the affected sites were floor of mouth (22 cases) and tongue (8 cases). Clinical findings were nonspecific, and prognosis was good following complete excision. Histology indicated that squamous, respiratory and/or gastrointestinal epithelium consistiuted basic structure of teratoid cyst in addition to simple cuboidal epithelium in 8 cases. Antibody against AE1/AE3 was strongly expressed and CK16 was weak in four types of epithelial lining of oral teratoid cyst. Expression of AE1, CK7, 8/18, 19 varied in superficial, suprabasal and basal cells of squamous epithelium but were strong in respiratory, gastrointestinal and simple columnar epithelium; only gastrointestinal epithelium expressed CK20 heterogeneously.

CONCLUSIONSOral teratoid cyst showed the highest incidence in children, and floor of mouth and tongue were mostly affected sites. Features of histology were complex, and immunohistochemistry indicated that epithelium of oral teratoid cyst shared similar patterns of cytokeratin with counterpart of normal tissues, showing different origin and differentiation of epithelial lining of the present cyst.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Humans ; Immunohistochemistry ; Infant ; Keratins ; metabolism ; Male ; Mouth Neoplasms ; metabolism ; pathology ; Retrospective Studies ; Teratoma ; metabolism ; pathology ; Young Adult

OBJECTIVETo investigate the resection range of mesorectum and rectum below the inferior margin of tumor for the total mesorectum excision (TME) in middle-low rectal cancer.

METHODSSixty patients were enrolled in the study. After TME operation, serial 5 mm interval sections were made in specimens of middle-low rectal cancer. The retrograde metastasis of rectal cancer was observed by routine HE staining.

RESULTSThe phenomena of retrograde metastasis in mesorectum were found in 15 cases, and the distance of retrograde metastasis was 0.5-4.0(2.47+/-1.06) cm, which was correlated with Dukes stage, lymph node metastasis and histological differentiation. The retrograde metastases in bowel were found in 11 cases, and the distance of retrograde metastasis was 0.5-4.0 (1.64+/-1.16) cm, which was correlated with histological differentiation.

CONCLUSIONSThe distal mesorectum should be resected at least 4 cm when TME is carried out, and the distal bowel at least 2.5 cm. More than 5 cm mesorectum and bowel should be resected when advanced Dukes stage, extensive lymph node metastasis and poor histological differentiation occurred.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Rectal Neoplasms ; pathology ; Rectum ; pathology

OBJECTIVETo make prenatal dignosis of X-linked adrenoleukodystrophy (ALD) for the prevention of the disease.

METHODSEighteen amniocenteses were performed on 17 suspected carriers of X-ALD during 18-30 gestation weeks. The very long chain fatty acids (VLCFAs) levels of cultured amniocytes were tested by gas chromatography-mass spectrometry (GC/MS). The plasma VLCFAs levels were measured in 8 of the 18 prenatal diagnosed children when they were born or after abortion. ABCD1 gene mutation analysis was carried out in 8 cases by PCR and sequencing. ALDP of amniocytes was tested by Western blotting in 2 cases from a family, one female, another male, and the VLCFAs of cultured amniocytes were increased in both of them.

RESULTSAmong the 18 fetuses, 10 were males and 8 were females. The VLCFAs levels of the cultured amniocytes were increased in 3 males and 4 females. The postnatal plasma VLCFAs were normal in 5 cases with normal VLCFAs levels of amniocytes, and increased in 3 cases with high VLCFAs levels of amniocytes. ABCD1 gene mutations were found in 4 cases with high VLCFAs levels of amniocytes, no mutation was found in other 4 cases with normal VLCFAs levels of amniocytes. ALDP of amniocytes could be detected in the female with high VLCFAs levels of amniocytes, and it could not be detected in the male with high VLCFAs levels of amniocytes. Three male fetuses with high VLCFAs levels of amniocytes were aborted. The others who were born were normal clinically so far.

CONCLUSIONThe prenatal diagnosis is very important for the prevention of ALD. Amniocyte VLCFAs level analysis combined with ABCD1 gene mutation analysis and ALDP test could make a proper prenatal diagnosis.

ATP Binding Cassette Transporter, Sub-Family D, Member 1 ; ATP-Binding Cassette Transporters ; genetics ; Adrenoleukodystrophy ; diagnosis ; genetics ; metabolism ; Adult ; DNA Mutational Analysis ; Fatty Acids ; metabolism ; Female ; Gas Chromatography-Mass Spectrometry ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Prenatal Diagnosis ; methods

OBJECTIVETo evaluate the influence of Fuzhenghuayu decoction on fibrotic liver tissue and activated hepatic stellate cells (HSCs) using a carbon tetrachloride (CCl4)-induced liver cirrhosis rat model system.

METHODSSixty-four Sprague-Dawley rats were randomly divided into the following groups: normal (non-model, non-drug intervention), CCl4 liver fibrosis model, and CCl4 liver fibrosis model Fuzhenghuayu drug intervention at low dose (0.75 g/kg/d) and high dose (1.5 g/kg/d). The drug intervention was administered via oral-gastric irrigation once daily for 6 times per week over a 6-week period. Four rats from each group were sacrificed at the end of week 2, 4, and 6 for serum and liver tissue collection. Liver fibrosis was evaluated by histology, and expression of a-smooth muscle actin (a-SMA) was determined by immunohistochemistry. Liver function was assessed by measuring levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin (TBil). Between-group comparisons were made by completely random design and ANOVA with Bonferroni correction.

RESULTSAt the end of weeks 2, 4 and 6, all four groups showed significantly different levels of ALT, AST, and TBil; in addition, the model group and drug intervention groups had significantly higher levels of ALT, AST, and TBil than the control group, the drug intervention groups showed significantly lower levels of ALT, AST, and TBil than the model group (P less than 0.01 or less than 0.05), and the differences between the low dose and high dose groups reached statistical significance (P less than 0.01 or less than 0.05). At the end of weeks 2, 4 and 6, the model group and drug intervention groups had significantly higher area ratio of liver fibrosis than the normal group (F = model: 18.68, low dose: 49.95, high dose: 82.44, P less than 0.01), but the two drug intervention groups had significantly less area ratio of liver fibrosis than the model group (P less than 0.05) and the high dose group showed the most robust decrease. In addition, the model group and drug intervention groups showed higher expression of a-SMA than the normal group (F = model: 18.68, low dose: 49.95, high dose: 82.44, P less than 0.01), but two drug intervention groups had significantly less a-SMA than the model group (F = model: 46.32, low dose: 40.30, high dose: 58.42, P less than 0.05) and the high dose group showed the most robust decrease.

CONCLUSIONThe Fuzhenghuayu decoction reduces the numbers of activated HSCs, thereby leading to down-regulated a-SMA expression and reduced degree of liver fibrosis; these effects may represent the mechanism by which this drug suppresses hepatic fibrosis.

Actins ; metabolism ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Hepatic Stellate Cells ; drug effects ; Liver ; drug effects ; pathology ; Liver Cirrhosis, Experimental ; pathology ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the clinicopathologic and immunohistochemical features, biological behavior, diagnosis and treatment of solid pseudopapillary tumor of the pancreas (SPTP).

METHODSA retrospective clinical and clinicopathologic analysis was made on 33 cases of SPTP admitted from May 2001 to 2010 July. There were 7 male and 26 female patients, aging from 13 to 66 years with a mean of 34.3 years.

RESULTSThe tumor was located in pancreatic head of 10 patients, in pancreatic neck of 5 patients, in pancreatic body and tail of 18 patients. Of the 33 patients treated with surgery, 8 underwent simple resection of pancreatic tumor, 6 underwent pancreaticoduodenectomy, 3 underwent tumor resection plus pancreaticojejunostomy, 1 underwent tumor resection plus pancreaticogastrostomy, 11 underwent distal pancreatectomy, 4 underwent distal pancreatectomy plus spleen resection (1 underwent mesohepatectomy for hepatic metastasis). Sixteen of the 33 operations were completed by laparoscopy. Histologically, tumors were composed of papillary and microcystic solid structures, with uniformed population of cells. The pancreas and blood vessels invasion were identified in 3 cases, one of them was combined with liver metastasis, and they are male. Immunohistologically, the tumors were positive for α1-antitrypsin, α1-antichymotrypsin, β-catenin, CD10, CD56 and vimentin (all cases), neuron-specific enolase (3 cases), synaptophysin (6 cases), chromogranin A (4 cases), progesterone receptor (28 cases), estrogen receptor (3 cases), S-100 (6 cases). Totally 33 cases were followed up with a median period of 49 months without tumor recurrence.

CONCLUSIONSSPTP is of low graded malignancy. It primarily affects young women. It may be located in any part of pancreas. Immunohistochemistry is very important for the diagnosis and differential diagnosis of SPTP. Surgical resection is recommended as the treatment of choice. Laparoscopic distal pancreatectomy or tumor resection is feasible and safe for some selected patients, and the prognosis is good.

Adolescent ; Adult ; Aged ; Carcinoma, Papillary ; diagnosis ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; diagnosis ; pathology ; surgery ; Retrospective Studies ; Young Adult

OBJECTIVETo study the differential microRNAs expression between patients with essential hypertension and healthy controls.

METHODSWhole blood from 15 hypertensive patients and 5 controls healthies were separated into plasma at 3000 rpm for 10 minutes. MicroRNAs were harvested using kit, and stored at -80°C. MicroRNAs profiling were performed using Exiqon microRCURY(TM) LNA microRNAs array, and were quantitative RT-PCR for the differential microRNAs expression. In addition, we used a set of plasma samples from 24 hypertensive patients and 22 healthy donors to independently validate the expression of these signature microRNAs.

RESULTSMicroRNAs expression profile was found to be differentially in the essential hypertensive patients compared with the healthy donors. Of 1700 microRNAs detected on the microarray, 46 microRNAs were found to be differentially expressed in the essential hypertensive patient, 27 microRNAs were collected in Sanger microRNAs data-bank, the function of remaining 19 microRNAs were unknown. In the 27 microRNAs, 9 microRNAs were up-regulated in the hypertension patient samples, while 18 known microRNAs were down-regulated. MiR-296-5p (Fold change 0.47, P = 0.013) and miR-133b (Fold change 0.57, P = 0.033) were consistently down-regulated in the patient plasma, whereas let-7e (Fold change 1.62, P = 0.009) and hcmv-miR-UL112 (Fold change 2.72, P = 0.004), one human cytomegalovirus encoded microRNAs, were up-regulated in the patient samples. The microRNAs expression was independently validated using another sample. We showed that MHC class I polypeptide-related chain B (MHC class I polypeptide-related chain B, MICB) and Interferon regulatory factor 1 (Interferon regulatory factor 1, IRF1) were functional targets of hcmv-miR-UL112 by fluorescent reporter assays.

CONCLUSIONSThe hypertensive patients have distinct microRNAs expression Profile. Hcmv-miR-UL112 may have important implications toward pathogenesis of essential hypertension.

Adult ; Case-Control Studies ; Female ; Gene Expression Profiling ; Humans ; Hypertension ; blood ; genetics ; Male ; MicroRNAs ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Transcriptome

As an extension of the structure-based drug discovery, fragment-based drug discovery is matured increasingly, and plays an important role in drug development. Fragments in a small library, with lower molecular mass and high "ligand efficiency", are detected by SPR, MS, NMR, X-ray crystallography technologies and other biophysical methods. Then they are considered as starting points for chemical optimization with the guidance of structural biology methods to get good "drug-like" lead and candidate compounds. In this article, we reviewed the current progress of fragment-based drug discovery and detailed a number of examples to illustrate the novel strategies.
Computer-Aided Design ; Crystallography, X-Ray ; Drug Discovery ; methods ; Ligands ; Magnetic Resonance Spectroscopy ; Peptide Fragments ; chemical synthesis ; chemistry ; Protein Conformation ; Small Molecule Libraries ; Surface Plasmon Resonance