Objective To establish a micellar electrokinetic capillary chromatography method for content determination of geniposide in Biyuanshu Oral Liquid. Methods Acetaminophen was used as an internal standard, and the separation was performed on an uncoated fused silica capillary of 52 cm × 50 μm ID (42 cm effective length) with the separation voltage of 25.0 kV. The running buffer contained 50 mmol/L borax, 100 mmol/L sodium dodecyl sulfate and 15％ acetonitrile (pH=10). The sample was injected by pressure (10 s, 0.5 psi) and detected at 238 nm. Results Geniposide was in good linearity range of 15.02–320.48 μg/mL (r=0.9995). The repeatability (low, medium and high concentration of samples) and intermediate precision assays gave satisfactory RSD values of less than 1.77％and 2.01％, respectively. The average recovery of geniposide in Biyuanshu Oral Liquid was 97.50％ and the RSD was 4.43％. The contents of geniposide determined by micellar electrokinetic capillary chromatography were in accordance with the results of HPLC analysis. Conclusion The method is simple, fast, accurate and precise, which can be used for the content determination of geniposide in Biyuanshu Oral Liquid.

OBJECTIVETo determine the correlation between the expression of CARMA1 mRNA and MUM1 protein, as well as its effects on clinicopathological features and prognosis of diffuse large B cell lymphoma (DLBCL).

METHODSThe immunophenotype (CD20, CD79a, CD10, MUM1, Bcl6) and proliferation index of DLBCL cells were examined by immunohistochemistry (IHC). CARMA1 mRNA was detected by RT-PCR.

RESULTSCARMA1 mRNA was detected in 76 of 89 (85.40%) cases with DLBCL. The level of CARMA1 mRNA was higher in MUM1-postive group than in MUM1-negative group. No correlation was found in the expression intensity between the two molecules (P = 0.084). Ki67 positive rate was higher in MUM1(+) cases than in MUM1(-) ones (P = 0.030). There was no difference between MUM1(+) and MUM1(-) cases in sex, median age, staging, primary site and other clinicopathological features. In 58 CARMA1 mRNA positive cases, low expression cases showed more in earlier stage and more males. No difference in survival status was identified between cases with and without MUM1 expression, over- and low-expression of CARMA1 mRNA, as well as over- and low-expression of CARMA1 mRNA among 58 cases with MUM1 expression.

CONCLUSIONThe expression of CARMA1 mRNA is likely associated with the expression of MUM1 and shows male predominance in DLBCL. The expression of CARMA1 may be involved with pathogenesis and progression of ABC-like DLBCL. The two molecules correlated somewhat with some clinicopathological features, but not with survival of DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; CARD Signaling Adaptor Proteins ; genetics ; metabolism ; Chromosomal Proteins, Non-Histone ; genetics ; metabolism ; Female ; Guanylate Cyclase ; genetics ; metabolism ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; metabolism ; pathology ; Neoplasm Staging ; Young Adult

To produce antibodies capable of neutralizing botulinum neurotoxin type B(BoNT/B),We cloned the carboxy-terminal end of Hc containing the major determinants responsible for specific toxin,induced and purifed.The heavy-chain and kappa light-chain variable region gene repertoire of immunoglobulin were amplified individually from the spleen cell mRNA by RT-PCR and joined as a single-chain Fv(scFv)DNA fragment.These fragment were cloned into the phagemid pCANTAB5E and the phage display library was constructed.Results showed that the high affinity scFv was obtained after 4 rounds of panning,with its DNA sequence conforming to that of mouse antibody.

OBJECTIVETo create a stable and reliable model of skin avulsion in rats.

METHODS30 male, SD rats were randomly divided into axial pattern skin flap (9 cm x 3 cm) group and random pattern skin flap group (6 cm x 4 cm), each having the control groups and avulsion groups. Flaps were subjected to avulsion force of 6 kg in axial pattern skin flaps or 8 kg in random pattern skin flaps, and the lasting time was 8 s or 12 s, respectively. Retraction of wounds and necrosis of skin flaps were observed.

RESULTSThere was more significant wound retraction in avulsion groups than that in control groups on post-operation day 7 (P < 0.05). The proportion of the wound retraction increased by 1 fold in avulsion groups on post-operation day 14 as compared to post-operation day 7 (P < 0.01). Interestingly, necrosis of partial or most of skin flaps was observed in all animals of avulsion groups, while slight necrosis happened in one of six in control animals. The necrosis area of flaps was 38% - 77% when avulsed for 8 s, and was 40% - 80% when avulsed for 12 s in axial pattern skin flaps. However, the necrosis area in random pattern skin flaps was smaller than that in axial pattern skin flaps, from 17% - 40% when avulsed for 8 s to 24% - 43% when avulsed for 12 s.

CONCLUSIONSIt might be possible to create animal model of skin avulsion injuries with rats.

Animals ; Disease Models, Animal ; Lacerations ; Male ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; Surgical Flaps

To study the pharmacokinetics of cantide, an antisense oligonucleotide, and its metabolites after iv gtt administration in rhesus monkeys, a dual solid phase extraction pretreatment method coupling with non-gel sieving capillary electrophoresis analysis method was used for determination of cantide and its metabolites in plasma and their pharmacokinetic parameters were calculated. The pharmacokinetic behavior of cantide and its metabolites (M1 and M2) after iv gtt administration (8, 16 and 24 mg kg(-1)) in rhesus monkeys were investigated. After iv gtt administration of cantide to rhesus monkeys, cantide in plasma was eliminated rapidly and the terminal elimination half-life (t1/2) was 57.91-77.97 min, the correlation coefficients (r) to the dose of Cmax AUC(o-inf) and AUC(0-t) of the prototype was 0.9918, 0.9568 and 0.9773, respectively. The metabolites of cantide reached the Cmax following cantide immediately and the Cmax of metabolites were lower than that of the prototype. The CL(S) of cantide and its metabolites (M1 and M2) were 1.60-2.19, 5.92-8.58 and 6.07-8.78 mL min(-1) kg(-1), respectively. So, it is concluded that the Cmax of cantide and its metabolites increased with the dose, which is the same as their AUC(0-inf) and AUC(0-t). The CL(S) of metabolites were higher than that of the prototype. The MRT and t1/2 of metabolites in the high dose group increased obviously.
Animals ; Area Under Curve ; Electrophoresis, Capillary ; methods ; Female ; Half-Life ; Infusions, Intravenous ; Macaca mulatta ; Male ; Oligonucleotides, Antisense ; blood ; metabolism ; pharmacokinetics ; Phosphorothioate Oligonucleotides ; blood ; metabolism ; pharmacokinetics ; Solid Phase Extraction

OBJECTIVETo observe the peripheral neurotoxicity of 1-bromopropane (1-BP) by developing an animal model of peripheral neuropathy through oral administration of 1-BP.

METHODSForty male Wistar rats were randomly and equally divided into low-dose group (200 mg/kg), medium-dose group (400 mg/kg), high-dose group (800 mg/kg), and control group. The rats in the low-dose, medium-dose, and high-dose groups were orally given 1-BP (dissolved in corn oil), while the rats in the control group were orally given an equal volume of corn oil. The oral administration (0.2 ml/100 g BW) was performed once per day, 5 days per week, for 16 consecutive weeks. Neurobehavioral indices including gait score, hindlimb grip strength, and hindlimb landing foot splay were recorded periodically. Hematological and biochemical parameters were also measured during and after 1-BP exposure.

RESULTSThe gait scores were significantly higher in the high-dose group (after 8 ∼ 16 weeks of 1-BP exposure), medium-dose group (after 14 ∼ 16 weeks of 1-BP exposure), and low-dose group (after 15 ∼ 16 weeks of 1-BP exposure) than in the control group (P < 0.05, P < 0.01). Compared with the control group, the high-dose group showed significantly decreased hindlimb grip strength after 9, 12, and 14 weeks of 1-BP exposure (P < 0.05, P < 0.01), with the hindlimbs paralyzed after 16 weeks of 1-BP exposure. After 16 weeks of 1-BP exposure, the hindlimb grip strengths of rats in the medium-dose and low-dose groups were decreased to 72.6% and 91.2% of the control value (P < 0.01, P < 0.05). Compared with the control group, the high-dose group showed significantly increased hindlimb landing foot splay after 12, 14, and 16 weeks of 1-BP exposure, and the medium-dose group showed significantly increased hindlimb landing foot splay after 14 and 16 weeks of 1-BP exposure (P < 0.05, P < 0.01). The high-dose and medium-dose groups showed significantly higher serum alanine aminotransferase (ALT) activity than the control group after 8 weeks of 1-BP exposure, and so did the low-dose group after 16 weeks of 1-BP exposure (P < 0.01).

CONCLUSIONThe nervous system is sensitive to the toxic effect of 1-BP, and 1-BP exposure can induce peripheral neuropathy in rats.

Animals ; Disease Models, Animal ; Hydrocarbons, Brominated ; administration & dosage ; toxicity ; Male ; Peripheral Nervous System Diseases ; chemically induced ; physiopathology ; Rats ; Rats, Wistar

The aim of this study was to investigate the expression of histone demethylase lysine specific demethylase1 (LSD1) in patients with acute leukemia (AL) and its clinical significance. LSD1 protein expression level was detected by semi-quantitative Western blot in HL-60 and SHI-1 leukemia cell line, in bone marrow mononuclear cells of acute AL patients with different condition [new diagnosis, complete remission (CR) and relapse] and in patients with non malignant hematopathy (control). Clinical data of AL patient followed up was collected. The relationship of LSD1 expression level with clinical prognosis was analyzed. The results showed that in HL-60 and SHI-1 leukemia cell line, LSD1 expression was strong positive, relative amount (LSD1/β-actin gray level rate) was 4.647 ± 3.840 and 1.628 ± 0.185 (n = 4) respectively. In 72 AL patients, LSD1 expression levels were quite different. LSD1 positive rate was 56.9% (41/72), average relative amount was 1.053 ± 1.976. In 17 controls, LSD1 positive rate was 0%, relative amount was 0.004 ± 0.012. The LSD1 positive rate in newly diagnosed AML or ALL group (90.4%, 77.8%) and refractory/relapse AML or ALL group (100%, 100%) was higher than that in AML or ALL CR group (4.7%, 0%) (p = 0.000), relative amount of LSD1 showed no statistically difference between newly diagnosed AML and ALL groups (1.177 ± 1.646, 1.275 ± 1.845) or refractory/relapse group (2.050 ± 2.470, 4.107 ± 3.676) and CR group (0.029 ± 0.033, 0.019 ± 0.024) (p > 0.05). In all AL patients, LSD1 positive rate in newly diagnosed (84.6%) and refractory/relapse groups (100%) was higher than that in CR group (3.8%). LSD1 relative amount in newly diagnosed group (1.274 ± 1.760), refractory/relapse group (3.359 ± 3.319) and CR group (0.027 ± 0.031) was higher than that in control group (p < 0.01), and in refractory/relapse group was higher than that in newly diagnosed group and CR group (p < 0.01), in newly diagnosed group was higher than that in CR group (p < 0.01). It is concluded that overexpression of LSD1 is correlated with refractory or relapse in AL. LSD1 expression level can reflect disease status of AL patients and may be a predictive biomarker for unfavourable prognosis of AL.
HL-60 Cells ; Histone Demethylases ; metabolism ; Humans ; Leukemia ; metabolism ; Leukemia, Myeloid, Acute ; metabolism ; Recurrence

Adult ; Hepatic Encephalopathy ; therapy ; Hepatitis ; complications ; therapy ; Humans ; Liver Failure, Acute ; therapy ; Liver, Artificial ; Plasma Exchange

OBJECTIVETo clarify the gene type of Orientia tsutsugamushi (Ot) from Shandong province.

METHODSNested-polymerase chain reaction (nPCR) was used to identify the gene type of 23 isolated Ot strains, 2 pools of homogenized leptotrombidium (L.) scutellare, 10 blood specimens of scrub typhus patients, and at the same time to compare with the international reference strains Gilliam, Karp, Kato. Sequencing analysis of the Sta56 gene was also used to further identify the precise gene types.

RESULTSOf the 35 samples, 33 had the same products in the amplification of template Ot-DNA. They all belonged to Kawasaki strains endemic in Japan while 2 (FXS4 and LHGM2 strain) belonged to Karp strains. The Sta56 gene sequence homologies to Japan Kawasaki strain of the 2 representative strains (B-16 and FXS2 strain) of the 33 samples were 94.22%, 95.21% respectively, but they were less than 75.87% to other prototype strains; The homologies to Karp strain of FXS4 and LHGM2 strain were 83.03%, 96.45% respectively. B-16 and FXS2 strain were designated as of types strain Japan Kawasaki, FXS4 and LHGM2 as Karp strain.

CONCLUSIONThe results indicated that the dominant Ot strains in Shandong Province were similar to Kawasaki strains, but Karp strains also existed.

Animals ; DNA, Bacterial ; genetics ; Genotype ; Humans ; Mice ; Orientia tsutsugamushi ; genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Scrub Typhus ; epidemiology ; microbiology ; Sequence Homology ; Serotyping

OBJECTIVETo explore the clinical value of 64-slice spiral 3-phase CT enhanced scanning for preoperative TNM staging assessment of gastric carcinoma.

METHODSA retrospective study was performed to review the 64-slice spiral 3-phase CT enhanced scanning of 120 patients with gastric cancer diagnosed by biopsy prior to operation and postoperative pathological reports. All the findings were reviewed by two senior radiologic diagnosticians separately and compared with pathological findings.

RESULTSThe accuracy of 64-slice spiral CT enhanced scan was 79.2%(95/120) for T staging, 66.7%(10/15) for T1, 66.7%(14/21) for T2, 84.0%(42/50) for T3, and 85.3%(29/34) for T4. For gastric wall with single layer and multiple layers, the accuracy of CT enhanced scanning was 59.4%(19/32) and 81.8%(72/88) for T staging, and the difference was statistically significant(P<0.05). The accuracy of 64-slice spiral CT enhanced scan was 73.9%(85/115) for N staging, 75.5%(37/49) for N0, 70.3%(26/37) for N1, 75.9%(22/29) for N2. The accuracy of 64-slice spiral CT enhanced scanning was 89.2% for M staging.

CONCLUSION64-slice spiral CT 3-phase enhanced scanning can monitor the invasion, lymphatic metastasis, and distant metastasis of gastric cancer dynamically, which may become an important examination item for the preoperative evaluation of gastric cancer.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Retrospective Studies ; Stomach Neoplasms ; diagnostic imaging ; pathology ; Tomography, Spiral Computed ; methods