In order to compare the differences of 35 Menthae Herba samples collected on the market and at producing areas, the contents of six total terpenoids, the essential oil and chromatographic fingerprints were analyzed, which provided evidences for drawing up the commodity specifications and grading criteria of Menthae Herba. GC-MS method was used to analyze the chemical constituents of 35 different samples. The chromatographic fingerprints obtained by using GC were then evaluated by similarity analysis, hierarchical clustering analysis and principal component analysis. The relativity between the content of six terpenoids and the essential oil were studied. In this study, the chemical profiles of 35 samples from different producing areas had significant disparity. All samples collected in the report could be categorized into four chemical types, L-menthol, pulegone, carvone and L-menthone, but the chemical profiles had no relationship with the areas. The chromatographic fingerprints of the samples from different types were dissimilar, while the different producing areas were difficult to be separated. It was indicated that the content of volatile oil was positively correlated with the content of L-menthol and the sum of six total terpenoids. The content of the essential oil, L-menthol and the sum of six total terpenoids of Menthae Herba were considered as one of the commercial specifications and grading criteria. These results in the research could be helpful to draw up the commercial specification and grading criteria of Menthae Herba from a view of chemical information.
Cluster Analysis ; Gas Chromatography-Mass Spectrometry ; Mentha ; chemistry ; Oils, Volatile ; analysis ; Principal Component Analysis ; Terpenes ; analysis

creatinine concentration. Conclusion Bisphosphonates can decrease serum total calcium levels in hypercalcemia crisis caused by PHPT effectivelywith mild adverse events.

Objective To investigate the clinical significance of a new method,axis-line-distance technique(ALDT),in scoliosis measurement.Methods Thirty cases with idiopathic scoliosis were measured on two separate occasions by six observers with the Cobb technique and the ALDT on PACS workstation.The interval time between two measurement occasions were three weeks.The data were analyzed statistically with the paired-sample t-test.Results(1)Concerning intraobserver variance in two measurement occasions,the minimum variance,the maximum variance and the average variance were 0, 24.00?,5.71??1.54?for Cobb technique and 0,12.00 mm,(1.95?0.58)mm for ALDT.There were significant measurement differences for four observers with Cobb method 39.00??10.69?versus 36.50?? 10.63?,31.73??10.96?versus 37.30??9.65?,32.03??7.49?versus 27.86??9.00?,29.77??8.87? versus 34.20??7.26?,all P0.05].(2)Concerning interobserver variances in six observers,the average measurement variance was 5.07??0.35?for Cobb method,and(2.32?0.26)mm for ALDT. There were significant measurement differences for every observer using with Cobb method(36.63??10.30? versus 33.27??10.10?,39.00?10.69?versus 31.73??10.96?,32.03??7.49~ versus 29.78?? 8.87?,36.63??10.30?versus 39.00??10.69?versus 32.03??7.48?,39.00??10.69?versus 32.03?? 7.49?,all P

HBV-associated acute-on-chronic liver failure is prevalent in mainland China.The prognosis of HBV-ACLF is poor.The mortality of HBV-ACLF is approximately 80％.Therefore,a prognostic indicator was needed in order to allow us to intervene as soon as possible.The model for end-stage liver disease (MELD) scoring system is widely used to predict the prognosis of liver failure.However,the assessment is too complex to restrict its application.This study aimed to investigate the expression ofIP-10 in peripheral blood mononuclear cells (PBMC),in order to explore the relationship between the expression and prognosis of patients with HBV-ACLF.The mRNA level of IP-10 in PBMCs were analyzed in 80 patients with HBV-ACLF,40 patients with chronic hepatitis B (CHB)and 40 healthy people by fluorescent quantitative PCR.IP-10 mRNA level was significantly higher in the HBV-ACLF group than in the other two groups (P＜0.01).Group with MELD score below 30 had lower IP-10 mRNA level than group with MELD score over 30 (P＜0.05).The IP-10 mRNA level in PBMCs in positive group was higher than that in negative group (P＜0.01).With a threshold of 0.925,the area under the receiver operating characteristic (ROC) curves was 0.815.These findings suggest that assessment of IP-10 mRNA level in the PBMCs would be helpful for evaluating the disease severity and prognosis in patients with HBV-ACLF.

OBJECTIVE: To investigate the role of oxidative stress in acute liver injury during crushing hindlimbs in rabbit. METHODS: The crushing injury model in rabbit was established by intermittent crushing the hind limbs of rabbit with standard weight. The ALT and AST activities were spectrophotometrically measured. The weight ratio (wet/dry,W/D) of livers was measured with scale, and the pathologic changes were observed. Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and total anti-oxidant capacity (T-AOC) as well as malondialdehyde (MDA) level were spectrophotometrically measured. RESULTS: As compared with control rabbits, crushing hindlimbs of rabbits induced acute liver injury with the increase in ALT and AST activities in serum,which were 4.31 (P < 0.01) and 10.54 times (P < 0.01) of control group respectively, there were cellular swellings and slight congestion of hepatic sinuses. In addition,crushing hind-limbs elicited significant decrease in SOD,CAT,GSH-Px activity and T-AOC to 17%, 29%, 24% and 21% (P < 0.01) compared with control group respectively, whereas MDA level markedly enhanced. CONCLUSION Crushing hindlimbs of rabbits induced acute liver injury and significant decrease in anti-oxidant capacity, the latter maybe play an important role in crushing hind-limbs of rabbits-elicited the acute liver injury.
Acute Disease ; Alanine Transaminase/blood* ; Animals ; Aspartate Aminotransferases/blood* ; Catalase/metabolism* ; Disease Models, Animal ; Female ; Glutathione Peroxidase/metabolism* ; Hindlimb/injuries* ; Liver/pathology* ; Liver Diseases/pathology* ; Male ; Malondialdehyde/metabolism* ; Oxidative Stress ; Rabbits ; Superoxide Dismutase/metabolism*

OBJECTIVE: To investigate the change of nitric oxide (NO) level in local muscles induced by crushing hind-limbs in rats. METHODS: The rat experimental model of hind-limb crushing injury was established by crushing the hind limbs of rats with standard weight for 5 hours, thereafter releving the standard weight for another 5 hours. The rats were randomly divided into sham group, crushing group, crushing and injecting aminoguanidine (AG) group, crushing and injecting L-arginine (L-Arg) group. The NOS activity and NO level in local muscles and serum were spectrophotometrically measured, and iNOS and eNOS protein expressions in local muscles were examined by immunohistochemistry. The weight ratio of wet to dry (W/D) of local muscles was measured and the pathologic changes were observed. RESULTS: The crushing hind-limbs induced serious primary and secondary injuries of local muscles such as rupture and rhadomyolysis of skeletal muscular fibers, interstitial vascular congestion and edema, and marked increase in W/D. The expressions of eNOS and iNOS were upregulated in local muscle in crush group compared with sham group. The NOS activity and NO level in local muscles and serum significantly increased. There was positive relationship between NO level and W/D in local muscles. With the usage of AG and L-arg, the hind-limb injuries seemed alleviated and aggravated, respectively. CONCLUSION The crushing hind-limbs of rats elicited the upregulation of eNOS and iNOS protein expression, the enhancement of NOS activity and the excess production of NO, the latter of which was involved in the mediation of secondary pathological changes in local muscles.
Animals ; Disease Models, Animal ; Female ; Hindlimb/injuries* ; Immunohistochemistry ; Male ; Muscle, Skeletal/pathology* ; Nitric Oxide/blood* ; Nitric Oxide Synthase/metabolism* ; Rats ; Rats, Wistar ; Soft Tissue Injuries/pathology*

OBJECTIVE: To compare the pathomorphologic changes between the pancreas in acute necrotizing pancreatitis (ANP) and that in acute deaths of rats (within 48 hours) so as to find the distinctions. METHODS: The animal models of ANP and other acute deaths (electroshock, mechanic asphyxia/strangle, and acute poisoning with tetramine) were established according to the criteria. Half-quantitative grading and image quantitative analysis methods were employed to observe the gross and microscopic changes of the pancreases. RESULTS: Three features including inflammation infiltrate, fat necrosis and calcium deposit in the ANP group were considerably different from that in other acutely died rat group (P<0.05). CONCLUSION Inflammation infiltrate, fat necrosis and calcium deposit are the most important pathologic features found in ANP by common light microscope, distinguishing ANP from postmortem pancreatic autolysis.
Animals ; Autolysis ; Female ; Forensic Pathology ; Male ; Pancreas/pathology* ; Pancreatitis, Acute Necrotizing/pathology* ; Poisoning/pathology* ; Postmortem Changes ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate whether three biallelic polymorphisms at the position -592, -819 and -1082 in the promoter region of the IL-10 gene were associated with the incidence of autoimmune liver disease.

METHODSThe IL-10 -592 and IL-10-1082 polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphisms analysis (PCR-RFLP), while polymerase chain reaction- sequence specific primer (PCR-SSP) assay was used to detect IL-10 -819 polymorphisms.

RESULTSAmong 54 Chinese patients with AIH or 77 Chinese patients with PBC versus healthy controls, the frequency of AA, GA genotypes at IL-10 gene promoter -1082 position was 87.0% or 83.1% versus 90.0%, 13.0% or 16.9% versus 10.0%, respectively (P > 0.05), the GG genotype in Chinese populations is absent; the frequency of CC, CT, TT genotypes at IL-10 gene promoter -819 position was 11.11% or 9.1% versus 8.1%, 44.4% or 53.3% versus 45.0%, 44.4% or 37.7% versus 46.9%, respectively (P > 0.05); the frequency of CC, CA, AA genotypes at IL-10 gene promoter -592 position was 4.9% or 14.3% versus 10.0%, 51.2% or 53.3% versus 51.9%, 43.9% or 32.5% versus 38.1%, respectively (P > 0.05). No alleles differed significantly in each groups.

CONCLUSIONThere were no association between IL-10 gene polymorphisms and autoimmune liver disease

Adult ; Aged ; Female ; Hepatitis, Autoimmune ; genetics ; immunology ; Humans ; Interleukin-10 ; genetics ; Liver Cirrhosis, Biliary ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Promoter Regions, Genetic ; genetics

OBJECTIVEThe goal of this study was to construct a eukaryotic expression plasmid containing the gene encoding herpes simplex virus type I glycoprotein D (HSV-1, gD) and evaluate its utility for DNA immunization in mice.

METHODSThe gD gene was amplified from viral DNA using PCR with EcoR I and BamH I restriction sites encoded on 5' and 3' ends, respectively. The PCR fragment was inserted into the transfer vector pGEM-T Easy. gD was then cut from this vector and inserted into the EcoR I and BamH I sites in the pcDNA3.1 at the multiple cloning sites (MCS). The recombinant plasmid, pcDNA3.1-gD1, was transfected into COS-7 cells using Lipofectamine according to the manufacture's instructions. The expression of the glycoprotein D was analyzed by immunoblotting of the cell lysates. 4-6 weeks old BALB/C mice were given two injections at tibia anterialis muscle, each containing 100 micrograms of plasmid DNA, on days 0 and 15. pcDNA3.1 was used as negative control. Blood samples were taken from all mice at weeks 0, 2, 4, and 6 after the first inoculation. Standard indirect ELISA was employed to evaluate the levels of specific total Ig in serum.

RESULTSThe recombinant plasmid was confirmed with restriction digestion and sequencing to contain target gene segment and expressed in COS-7 cells in vitro shown by Western blotting. The pcDNA3.1-gD1 immunized group induced specific antibody response as compared to the negative control, and the titer was about 1:2000.

CONCLUSIONSThe recombinant plasmid pcDNA3.1-gD1 is potential to be used as a candidate vaccine, for the treatment of HSV-1 infection.

Animals ; COS Cells ; metabolism ; Escherichia coli ; genetics ; Genetic Vectors ; Humans ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; biosynthesis ; genetics ; immunology ; Viral Envelope Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo explore the mechanism of immunomodulatory activity of triptolide on primary immune thrombocytopenia (ITP)patients-derived plasmacytoid dendritic cells (pDCs).

METHODSpDCs in peripheral blood of ITP patients before therapy (group 1), ITP patients in complete response (ITP-CR, group 2) and healthy donors (group 3) were sorted by flow cytometry, then incubated with triptolide at 0, 5, 10 or 30 µg/L. After 24 hours, we collected the supernatants and then detected the concentrations of IFN-α, IL-6 and TNF-α using ELISA. After 5 days, the cultured cells were collected and CD11c, CD80 and CD86 expressions of myeloid dendritic cells (mDCs) were analyzed by flow cytometry, the morphology of mDC was observed by light microscope and electron microscope.

RESULTSAfter incubation with triptolide at 10 µg/L, the levels of IFN-α, IL-6 and TNF-α in group 1 \[(451.32 ± 85.77) ng/L, (105.68 ± 23.85) ng/L and (135.78 ± 30.62) ng/L\] and group 2 \[(391.71 ± 72.49) ng/L, (84.73 ± 17.77) ng/L and (108.16 ± 23.21) ng/L\] were significantly higher than those in group 3 \[(335.51 ± 67.54) ng/L, (73.62 ± 21.82) ng/L and (95.58 ± 32.85) ng/L\] (all P < 0.05); the levels of IFN-α, IL-6 and TNF-α in group 1 were significantly higher than those in group 2 (all P < 0.05) in a dose-dependent manner (P < 0.05). CD11c, CD80 and CD86 expressions of mDC in group1 and group 2 were significantly higher than those in group 3 (all P < 0.05); CD11c, CD80 and CD86 expressions of mDC in group 1 were significantly higher than those in group 2 (all P < 0.05) also in a dose-dependent manner (all P < 0.05). Triptolide could inhibit pDCs from differentiation into mDCs, the latter displayed more immature morphology than untreated-pDCs.

CONCLUSIONTriptolide could decrease the immune function of pDCs from ITP, inhibit pDCs from differentiation and maturation.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Cell Differentiation ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; drug effects ; Diterpenes ; pharmacology ; Epoxy Compounds ; pharmacology ; Female ; Humans ; Male ; Middle Aged ; Phenanthrenes ; pharmacology ; Thrombocytopenia ; etiology ; immunology ; Young Adult