Objective To construct an electrochemical immunosensor for ultrasensitive determination of myeloperoxidase (MPO) .Methods The electrochemical immunosensor for M PO was prepared by modifying the electrode using Au-graphitized me-soporous carbon nanoparticles(AuNPs@ GMCs) hybrid and immobilizing MPO antibodies onto the glass carbon electrode surface . The effect of experimental parameters on the immunosensor and results comparison with ELISA were investigated .Results The immunosensor was sensitive to M PO with a linear relationship between 2 .000 and 300 .000 ng/mL and a correlation coefficient of 0 .999 ;the detection limit was 0 .5 ng/mL .The correlation coefficient of two methods was 0 .983 .Conclusion The immunosensor can be used for ultrasensitive detection of MPO .

Objective: To investigate the effect of moxibustion at Shenque (CV 8) on fatigue in rats with chronic exercise-induced exhaustion. Methods: Thirty male Sprague-Dawley (SD) rats were randomly divided into a blank group, a model group and a moxibustion group, 10 rats in each group. Except rats in the blank group, the remaining rats were subjected to create long-term exhaustion models by repeated swimming. After successful modeling, rats in the moxibustion group received mild moxibustion at Shenque (CV 8) for 15 min, once every other day with a total of 10 times. Rats in the model group and the blank group did not receive moxibustion. At the end of the treatment, the exhausted times, and the body weight of rats before and after the experiment were compared among groups. The levels of blood malondialdehyde (MDA) and urea nitrogen (BUN), as well as the activities of aspartate transarninase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) were also measured by the automatic biochemical analyzer, 24 h after the exhausting excise. Results: The 10th swimming time was significantly longer in the moxibustion group than that in the model group (P<0.01). The increase rate of the body weight was lower in the rats of the moxibustion group than that in the model group before the 7th and the 10th exhausting excise (P<0.05, P<0.01). The levels of serum MDA and BUN, as well as the activities of AST, ALT and LDH in the model group were higher than those in the blank group (all P<0.01). The levels of serum MDA and BUN, as well as the activities of AST, ALT and LDH in the moxibustion group were lower than those in the model group (P<0.01). Conclusion: Moxibustion at Shenque (CV 8) can decrease the serum levels of MDA and BUN, as well as activities of AST, ALT and LDH in the long-term fatigue rats, thus to improve the symptoms of fatigue.

OBJECTIVETo establish scientific and efficient quality control standard of Rhizoma et Radix Notopterygii through two quantitative parameters.

METHODThe contents of isoimperatorin and volatile oils in 29 commercial samples from 22 localities of Rhizoma et Radix Notopterygii were determined by HPLC and the method in the pharmacopoeia respectively. The data were statistically analyzed.

RESULTOnly 3 of 29 samples met the required standard in the Chinese Pharmacopoeia (2005 edition), 87% of samples newly produced in 2005 not met the standard. Statistics showed that the contents of volatile oils and isoimperatorin should reach 1.63% and 0.17% respectively when the samples meet requirement of criterion with more than 80% probability. The contents of volatile oil and isoimperatorin were not significantly different (P < 0.05) in the samples of 2005 and 2003, while the contents of isoimperatorin were significantly different between Notopterygium incisum and N. forbesii samples.

CONCLUSIONSince the criterion of volatile oils in the Chinese Pharmacopoeia (2005 version) for Rhizoma et Radix Notopterygii is too high to satisfy for most Rhizoma et Radix Notopterygii samples, it is suggested to reduce it to 1.6% (mL x g(-1)). The content of isoimperatorin is proposed to add to the quality standard of Rhizoma et Radix Notopterygii as above 0.2% (mg x g(-1)).

Apiaceae ; chemistry ; classification ; China ; Chromatography, High Pressure Liquid ; Furocoumarins ; analysis ; Oils, Volatile ; analysis ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Rhizome ; chemistry

OBJECTIVETo compare the effects of the point-to-point needling and perpendicular needling of auricular acupuncture for rapid analgesia on the patients with lumbar disc hernia, and find which method is better for this disease.

METHODSNinety two patients were paired according to the three factors as sex, age and degree of pain, and then randomly divided into the point-to-point needling group and perpendicular needling group. The methods of point-to-point needling through the skin and perpendicular needling towardS the skin were respectively applied at bilateral lumbar areas as auricular acupuncture, and added acupoints following the change of symptom. All patients accorded with the demands of McGill pain questionnaire (MPQ), before treatment, 5 and 30 min after treatment, the accumulated score of pain was recorded respectively for observing the difference of the effects in both groups.

RESULTSAfter 5 min treatment, the effect of analgesia on the male patients in the point-to-point needling group was better than that of in the perpendicular needling group (P<0.05), but there was no difference on the female patients in both groups (P>0.05). After 30 min treatment, the effect of analgesia in the point-to-point needling group was better than that of the perpendicular needling group (P<0.05). Besides the patients with light degree pain, there was no significant difference between the two groups after 5 min treatment as score of pain (P>0.05), paired t-test showed that the effects of analgesia on the patients with lumbar disc hernia in the point-to-point needling group was better than that of perpendicular needling group (P<0.05).

CONCLUSIONThe point-to-point needling method can rapidly relieve patients' lumbar disc hernia of pain, which is better than the perpendicular needling method.

Acupuncture Analgesia ; Acupuncture, Ear ; Adult ; Aged ; Female ; Humans ; Intervertebral Disc Displacement ; therapy ; Male ; Middle Aged ; Pain Management ; Pain Measurement ; Treatment Outcome ; Young Adult

Objective To establish HPLC analysis methods and fingerprint of different proportions of Angelicae Pubescentis Radix in Angelicae Sinensis Radix; To use HPLC fingerprint method to identify the adulteration of Angelicae Pubescentis Radix in Angelicae Sinensis Radix. Methods Agilent TC-C18 column (5 μm, 4.6 mm × 250 mm) was adopted. Acetonitrile (A) - 0.05% phosphoric acid (B) was the mobile phase, and the gradient elution (0–20 min, 95%–75%B; 20–60 min, 75%–65%B; 60–80 min, 65%–5%B) was used; The flowrate was 1 mL/min; Detection wavelength was 254 nm; Column temperature was 30 ℃. The HPLC spectra of Angelicae Sinensis Radix, Angelicae Pubescentis Radix and adulterated samples were compared and analyzed by using TCM chromatogram fingerprint similarity evaluation system. Results Angelicae Sinensis Radix and Angelicae Pubescentis Radix showed good chromatographic peaks in chromatogram separation. 11 peaks in HPLC fingerprint could be used as the mark, including 7th peak for ferulic acid, 32th peak for osthole, and 34th peak for columbianadin. Conclusion This method is rapid, simple and feasible, can fully reflect the quality of Angelicae Sinensis Radix and clearly identify whether Angelicae Pubescentis Radix is adulterated in Angelicae Sinensis Radix.

ObjectiveTo analyze Echinococcus infection in definitive and intermediate hosts in different zones of Qinghai plateau,Qinghai southern plateau,Qilian mountain-Hehuang valley and Chaidamu basin,and to provideascientificbasisfor developing controlstrategiesagainstEchinococcosisinfection. Methods Echinococcosis infection in definitive hosts,dogs and foxes,was identified by morphological observation; in domesticated and wild intermediate host animals was identified by anatomy and pathology; some of the suspected samples were further identified by molecular biological methods.ResultsStray dogs in different zones of Qinghai plateau were infected with Echinococcus granulosus,the infection rates were 38.71％(300/775),49.60％(124/250),and 9.76％(4/41 ) in Qinghai southem plateau,Qilian mountain-Hehuang valley and Chaidamu basin,respectively,and the difference was statistically significant(x2 =25.72,P ＜ 0.01 ).in addition,only Qinghai southern plateau dogs were infected with Echinococcus multiloularis,and the infection rate was 16.04％(98/611).The infection rates of fox with Echinococcus multilocularis were 22.89％(38/166) and 30.77％(12/39) in Qinghai southern plateau and Qilian mountain-Hehuang valley,respectively,and wolves were also found to be infected with Echinococcus granulosus in the same areas.The infection rates of domesticated sheep,yaks,goats and pigs with Echinococcosis were significantly different statistically in those different areas(x2 =82.70,41.82,212.63,194.58,all P ＜ 0.01 ).The infection rates of sheep and yaks were higher[43.43％(5664/13 042),49.47％(2917/5896),52.99％ (887/1674),42.18％ (779/1847),50.70％ (1049/2069),52.90％ (685/1295) ] in three areas.The infection rates of goats and pigs [3.26％ (7/215),0.00％ (0/108)] in Qinghai southern plateau were lower than that of other two areas[ 19.51％(119/610),26.91％(43/1598),47.91％(343/716),21.91％(71/324)].The infection rates of Ochotona curzoniae with Echinococcosis were 6.21％ (243/3910),1.80％ (3/167) and 0.00％ (0/199) in Qinghai southern plateau,Qilian mountain-Hehuang valley and Chaidamu basin,respectively,and the difference was statistically significant (x2 =18.50,P ＜ 0.01 ).Moreover,wild intermediate hosts of Echinococcosis,such as Microtus fuscus,Lepus oiostolus,Pseudois nayaur,Procapra picticaudata,and Prodorcas gutturosa were found to be infected only in Qinghai southern plateau.ConclusionsHuman is faced with a threat of Echinococcosis infection from various definitive hosts in different zones of Qinghai plateau.And stray dogs are the most crucial factor.The life-cycles of Echinococcus are very complicated in Qinghai plateau.Qinghai plateau is a key area in prevention and control of Echinococcosis infection in China.

OBJECTIVETo investigate the effect of Chinese recipe, Wuye Decoction (WYD), on immune function in patients with non-small cell lung cancer (NSCLC).

METHODSEighty-two patients of NSCLC with pathologically confirmed diagnosis, who had received operative treatment and completed the post-operational chemotherapy, were randomly assigned into 2 groups. Group A (42 cases) received WYD and Group B (40 cases) received no specific medicine. Positive rate of various peripheral lymphocyte subsets, including CD3, CD4, CD8, CD16, CD19 and CD25, in both groups was observed immediately after chemotherapy (T(0)) and 3 months later (T(1)), the same indexes of 20 healthy volunteers allocated in Group C were also determined at T(0) for control.

RESULTSThe positive rates of CD4, CD4/CD8, CD16, CD19 and CD25 were significantly lower (P < 0.05) while that of CD8 was significantly higher (P < 0.05) in Group A and B at T(0) than those in Group C; at T(1), these indexes, except CD25, got significantly restored in Group A with the level approaching normal range (P > 0.05), and showed significant difference from those in Group B (P < 0.05), since these indexes in that group remained unchanged at the corresponding period. As for CD25, it was insignificantly changed in Group A after WYD treatment, and thus, at T(1), it was still lower than that in Group C (P < 0.05) and showed insignificant difference as compared with that in Group B (P > 0.05). Comparison of CD3 among the 3 groups showed no significant difference (P > 0.05).

CONCLUSIONWYD could activate the immune function of NSCLC patients, and so it is recommended to be used in the treatment of NSCLC in clinical practice.

Adjuvants, Immunologic ; pharmacology ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; immunology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Lung Neoplasms ; drug therapy ; immunology ; Lymphocyte Subsets ; immunology ; Male ; Middle Aged ; Phenotype

OBJECTIVETo investigate the prevention and treatment of hepatitis B virus (HBV) reinfection and recurrence after orthotopic liver transplantation (OLT) for HBV related end-stage liver disease.

METHODSThe clinical data of 316 patients undergoing allograft orthotopic liver transplantation who lived more than 6 months and had HBV infection preoperative with complete data from March 2001 to March 2007 at the First Affiliated Hospital of Sun Yat-sen University. According to the HBV prevention strategy, these patients were divided into two groups: group with pure lamivudine (LAM) (n = 106) and group with lamivudine plus intramuscular injection of low dose anti-hepatis B immunoglobulin (HBIG) (n = 210).

RESULTSMean follow-up was 33.6 months. The rate of HBsAg negative conversion 1 week after OLT of the LAM group was 82.1% (87/106), significantly lower than that of LAM + HBIG group [91.0% (191/210), P < 0.05]. The rates of HBV reinfection, HBV recurrence, and YMDD mutation of the lamivudine group was 17.0% (18/106), 11.3% (12/106) and 8.5% (9/106) respectively, all significantly higher than those of LAM + HBIG group [6.2% (13/210), 3.8% (8/210) and 2.4% (5/210) respectively, P < 0.05 respectively]. All patients with HBV reinfection or HBV recurrence were treated with Adefovir, Entecavir or increased dose of HBIG and achieved better curative effect.

CONCLUSIONSThe therapy with high dose of HBIG combining with adefovir or entecavir is better for patients who have HBV reinfection. Patients with HBV recurrence after OLT should be administrated reasonable liver aid, immunity regulation and anti-hepatic fibrosis to obtain better transplant liver histological results and normal transplant liver function.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Hepatitis B ; drug therapy ; prevention & control ; Hepatitis B virus ; immunology ; Humans ; Immunoglobulins ; administration & dosage ; therapeutic use ; Lamivudine ; administration & dosage ; therapeutic use ; Liver Transplantation ; Male ; Middle Aged ; Postoperative Period ; Retrospective Studies ; Secondary Prevention ; Young Adult

OBJECTIVETo investigate the methods of culturing and identifying mouse myeloid semimature dendritic cell (smDC) in vitro.

METHODSMyeloid monocytes derived from 6-week-old C57 BL/6 mice were cultured in RPMI-1640 medium containing 10% fetal bovine serum, 2 ng/ml recombinant murine granulocyte macrophage-colony stimulating factor (GM-CSF), and 20 ng/ml recombinant murine interleukin (IL)-4 for 9 days. Then cells were incubated with 40 ng/ml tumor necrosis factor-alpha (TNF-alpha) for 24 hours to obtain smDC. Meanwhile, smDC was differentiated into mature dendritic cell (mDC) or immature dendritic cell (iDC) by treatment with 1 micro/m1 lipopolysaccharide (LPS) or without LPS. The morphological features of smDC were assayed by inverted microscopy and scanning electron microscopy. Surface markers such as CD11c, CD4O, CD8O, CD86, and MHC-II were tested by flow cytometry. IL-1beta, IL-6, IL-12, and IL-10 in the supernatant were tested by ELISA. The activation of allogene lymphocyte (BALB/c mice) stimulated by C57BL/6 myeloid smDC in mixed lymphocyte reaction was examined by Cell Counting Kit-8 in vitro.

RESULTSThe shape of smDC was round or oval-shaped, and the diameter of smDC was about 15 microm. The length of smDC dendrite was between 5 to 10 microm. smDC, iDC, and mDC all expressed high level of CD11 c. The expressions of MHC-II, CD40, CD80, and CD86 on smDC were higher than those of iDC and lower than those of mDC. IL-1beta, IL-6, and IL-12 secretion of smDC was significantly lower than that of mDC (P < 0.01), and IL-12 was significantly lower than that of iDC (P < 0.05), while no significant difference of IL-1beta and IL-6 secretion was found between smDC and iDC (P > 0.05). Furthermore, IL-10 secretion was not significantly different among these three kinds of DCs (P > 0.05). The effect of allogene lymphocytes activation on smDC was significantly lower than that of mDC and positive control (P < 0.01), but had no significant difference when compared with that of iDC and negative control (P > 0.05).

CONCLUSIONSsmDC may be a relatively independent dendritic cell sub-population in terms of function and morphology. It is a feasible way to induce myeloid monocytes to differentiate into smDC using GM-CSF, IL-4, and TNF-alpha in vitro.

Animals ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Cytokines ; immunology ; Dendritic Cells ; cytology ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Monocytes ; cytology ; immunology

OBJECTIVETo develop a LC-MS method for the determination of senkyunolide I (SI) in rat plasma, in order to observe whether there is significant change in the pharmacokinetics parameters of complex prescriptions of Huoluoxiaolingdan (HLXL) and single herbal extracts from Ligusticum chuanxiong Hort. in rats, and assess the effect of other components in HLXL on the pharmacokinetics of SI.

METHODTwelve male Sprague-Dawley (SD) rats were randomly divided into two groups, and orally administered with extract from HLXL and L. chuanxiong (both equal to SI 4.53 mg x kg(-1)). Their blood was collected at different time points for LC-MS, in order to detect the plasma concentration of SI. The pharmacokinetic parameters of SI were calculated by DAS 2.0 software. SPSS 16.0 software was used for independent-sample T-test and Nonparametric T-test.

RESULTA linear relationship of SI ranged from 6.750 to 675.0 microg x L(-1), and with the lowest limit of detection being 6.750 microg L(-1). Both of the plasma concentration-time curves of SI were fitted with the two-compartment model for extract of HLXL and L. chuanxiong. The detected AUC and Cmax of SI showed significant difference, with no significant difference in other parameters.

CONCLUSIONThe LC-MS determination method established in this experiment was so exclusive, accurate and sensitive that it is suitable for pharmacokinetic studies on extracts of HLXL and SI from L. chuanxion. The experiment results show that other ingredients of HLXL have noticeable effect on the absorption of SI in rat plasma.

Administration, Oral ; Animals ; Area Under Curve ; Benzofurans ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Male ; Metabolic Clearance Rate ; Random Allocation ; Rats ; Rats, Sprague-Dawley