To establish a method for determining the contents of six alkaloids (jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, palmatine hydrochloride, berberine hydrochloride) in six types of Coptidis Rhizoma pieces (crude pieces, ginger juice stir-fried pieces, vinegar stir-fried pieces, wine steamed pieces, wine stir-fried pieces, evodiae juice stir-fried pieces) by RP-HPLC, and explore the relationship with the curative effect of traditional Chinese medicine (TCM) and pharmacodynamics results. The chromatographic column was Welch XtimateTM C₁₈ (4.6 mm×250 mm, 5 μm), with 0.1% triethylamine solution (adjust pH at 10 with ammonium bicarbonate and ammonia) as mobile phase A and acetonitrile as mobile phase B for gradient elution (0-15 min, 10%-25%B; 15-25 min, 25%-30%B; 25-40 min, 30%-45%B) at a rate of 1.0 mL•min⁻¹. The column temperature was set at 30 ℃, and the wavelength was set at 270 nm. The six alkaloids showed a good linear relationship within the range of 0.85-16.96 mg•L⁻¹ (r=0.999 7), 1.25-24.96 mg•L⁻¹ (r=0.999 9), 2.05-40.96 mg•L⁻¹ (r=0.999 9), 3.65-72.96 mg•L⁻¹ (r=0.999 9), 2.88-57.60 mg•L⁻¹ (r=0.999 8), and 13.25-264.96 mg•L⁻¹ (r=0.999 6) respectively. The average recoveries (n=9) of the six alkaloids were 102.4% (RSD 1.2%), 101.8% (RSD 1.3%), 100.3% (RSD 1.8%), 100.7%(RSD 1.8%), 101.2% (RSD 1.5%) and 97.90% (RSD 2.0%) respectively, and their average contents were 3.55, 4.49, 9.12, 19.17, 15.69, 62.56 mg•g⁻¹, respectively. This determination method was accurate and repeatable, which could be used for the content determination in six types of Coptidis Rhizoma pieces. Data analysis on contents determination and preliminary pharmacodynamics results was conducted by using principal component analysis (PCA) and hierarchical clustering analysis (HCA). The analysis results showed that three types of Coptidis Rhizoma pieces (wine steamed pieces, wine stir-fried pieces, and evodiae juice stir-fried pieces) had significant differences with crude pieces, and the wine steamed Coptidis Rhizoma pieces showed most difference with crude pieces especially, mainly related to triglyceride (TG) and fasting blood glucose levels (FBG) in serum. In addition, columbamine hydrochloride was most affected among the six alkaloids. Those three types of Coptidis Rhizoma pieces (wine steamed pieces, wine stir-fried pieces, and evodiae juice stir-fried pieces), had more advantages for "anti-diabetes" in TCM clinical application, especially in the treatment of diabetic hyperlipidemia.

Objective: To evaluate the efficacy of allicin combined with cyclophosphamide on neuroblastoma (NB)-bearing mice and explore the immunological mechanism in it. Methods: A total of 30 NB-bearing mice were equally randomized into model group, cyclophosphamide group and combined therapy group, 10 nudemice were set as normal saline (NS) group. Cyclophosphamide group and combined therapy group were weekly injected with 60 mg/kg cyclophosphamide for four weeks; besides, combined therapy group was given with allicin (10 mg/kg/d) by gastric perfusion for 4 weeks; model group and NS group were given with the same volume of NS. Serum VEGF content was detected by ELISA pre-treating (0 d) and on the 3rd d, 14th d and 28th d; on 29th d, all mice were sacrificed and the tumor, liver, spleen and thymic tissues were weighted. Tumors were made into paraffin section for detecting tumor cell apoptosis and proliferation by TUNEL and BrdU method, respectively. Survival curves were drawn by Kaplan-Meier method. Results: After treatment, both treatment groups relieved on viscera indexes, VEGF level, T cell subsets distribution and tumor growth and each index of combined therapy group was better than cyclophosphamide group (P<0.05 or 0.01); only combined therapy group could significantly increase the lifetime of NB-bearing mice (μ ²=5.667, P=0.017). Conclusions: Allicin can improve T cell subsets distribution and inhibit VEGF expression through its immunomodulatory activity, thereby improve the efficiency on NB in coordination with cyclophosphamide.

Objective To study pharmacokinetic behaviors of the Chinese healthy volunteers and to evaluate the bioequivalence of two kinds of levetiracetam tablets.Methods Twenty healthy male volunteers were randomly divided into two groups, each group 10 cases, they were orally respectively administrated with single dose of treatment and reference preparations of levetiracetam ( each 0.5 g).The cleaning peri-od was one week.The concentration of levetiracetam in human plasma was measured by LC/MS/MS.Main pharmacokinetic parameters were calculated by using WinNonlin 6.3 software.Results Main pharmaco-kinetic parameters of test and reference preparation were as follow:t1/2 were(8.01 ±1.16), (8.02 ±0.93) h, tmax were (0.61 ±0.41), (0.76 ±0.61 ) h, Cmax were ( 13.65 ±2.63 ), ( 13.15 ±2.30 )μg? mL-1 , AUC0-t were ( 115.84 ±12.75 ) , ( 119.29 ±15.29 )μg? mL-1? h,AUC0-∞ were ( 121.44 ±15.17 ) , ( 124.89 ±16.73 )μg? mL-1? h,respectively.The relative bioavailability of test prepara-tion was ( 97.55 ±7.06 )%.Conclusion Two kinds of levetiracetam tablets are bioequivalent.

Objective To study pharmacokinetic behaviors of moxifloxa-cin in Chinese healthy volunteers and to evaluate the bioequivalence of 2 kinds of moxifloxacin hydrochloride tablets.Methods In randomized crossover study, 22 healthy male volunteers were given single oral dose of test and reference preparation of moxifloxacin tablet ( each 0.4 g ) .The concentration of moxifloxacin in human plasma was measured by LC/MS/MS.Main pharmacokinetic parameters were calculated by using WinNonlin 6.3 software.Results Main pharmacokinetic parameters of test and reference preparation were as follow: t1/2 were (14.91 ±2.56) and(14.68 ±2.02)h, tmax were (1.26 ±0.89)and (1.09 ±0.73) h, Cmax were (1914.84 ±461.20) and (1877.52 ±559.80) ng? mL-1, AUC0-t were ( 27406.96 ±7060.97 ) and ( 27414.48 ±6745.13 ) ng? mL-1? h, AUC0-∞ were ( 28361.29 ± 2409.57 ) and (28353.74 ±6883.95)ng? mL-1? h, respectively. The relative bioavailability of test preparation was(100.34 ±8.69)%.Conclusion Two kinds of moxifloxacin hydrochloride tablets are bioequivalent.

OBJECTIVETo explore an approach to rapidly and accurately identify the compounds as biomarkers of Chinese medicine (CM) syndromes.

METHODThe Fourier transform infrared (FT-IR) spectrometry was applied to investigate the characteristic components of a mice model of Kidney (Shen)-yang deficiency syndrome (KDS), and the remedial effect of a typical CM formula Shenqi Pill (). Thirty-six females and 18 males of Balb/c mice were randomly divided into KDS, Shenqi or control group. The females and males of the same group freely were mated for 96 h, and the males were taken out and only the female mice were raised. Females of the KDS group were threatened by a ferocious cat every other day for 14 d. After delivery, the KDS, or gestational threatened, offspring were raised at standard condition for 11 weeks. Then 10 male offspring were randomly selected, anaesthetized and their representative organs, i.e. testes, kidneys, lungs and feet were collected, for the FT-IR scan. Mice of the Shenqi group were intragastric administered Shenqi Pill; while mice in the KDS and control groups were given the same volume of saline.

RESULTSThe attenuated birth outcomes of the KDS group were displayed. The remarkable FT-IR differences of all organs between KDS mice and healthy control were mainly at 1,735-1,745 cm(-1) (indicating the increased levels of lipids) and at 1,640-1,647 cm(-1) and 1,539-1,544 cm(-1) (displaying the decreased proteins). No statistic FT-IR difference between Shenqi and control mice was observed.

CONCLUSIONIn accordance with major traits of KDS, prenatal stress extensively impaired the building up of proteins and resulting in the excessive lipid storage, and FT-IR could effectively identify the biomarkers of KDS.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Kidney Diseases ; drug therapy ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Spectroscopy, Fourier Transform Infrared ; methods ; Yang Deficiency ; drug therapy ; pathology

OBJECTIVETo investigate the changes in peripheral blood Th17 and CD4(+)CD25(+) regulatory T (Treg) cells and their significance among children with hand, foot and mouth disease (HFMD).

METHODSEighty-nine children with HFMD, including 55 cases of common HFMD and 34 cases of severe HFMD, were included in the study; and 30 healthy children were selected as the control group. The percentages of Th17 and CD4(+)CD25(+) Treg cells in CD4(+) T cells in peripheral blood were determined by flow cytometry. The expression levels of interleukin (IL)-10, transforming growth factor-β (TGF-β), and IL-17 were measured by enzyme-linked immunosorbent assay.

RESULTSCompared with the control group, the cases of common HFMD and severe HFMD had significantly increased levels of Th17 cells and IL-17 (P<0.05) but significantly decreased levels of CD4(+)CD25(+) Treg cells, IL-10, and TGF-β (P<0.05). The severity of the HFMD was positively correlated with the levels of Th17 cells and IL-17 in peripheral blood but negatively correlated with the levels of CD4(+)CD25(+) Treg cells, IL-10, and TGF-β.

CONCLUSIONSChildren with HFMD have increased response of Th17 cells but decreased response of CD4(+)CD25(+) Treg cells in peripheral blood. Th17/CD4(+)CD25(+) Treg cell imbalance may play an important role in the pathogenesis of HFMD.

Child ; Child, Preschool ; Hand, Foot and Mouth Disease ; immunology ; Humans ; Infant ; Interleukin-10 ; blood ; Interleukin-17 ; blood ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology ; Transforming Growth Factor beta ; blood

Objective To investigate the effects on ribbon synapse of inner hair cells by superpulsed CO2 laser-assisted cochleostomy in SD rats.Methods Eighteen SD rats were randomly divided into the laser-assisted surgery groups (2 W group and 5 W group),the sham-operated group and the control group.Ten of those were performed a cochleostomy using superpulsed CO2 laser with corresponding power.Auditory brainstem responses (ABR) were measured pre-and postoperatively.The ribbon synapses at apical and middle cochlear turns were observed under laser scanning confocal microscope and then were quantified with 3ds Max software.Results The postoperative ABR thresholds of the 2 W and 5 W groups were larger than the preoperative cases (t =-5.65,P ＜ 0.01 ; t =-4.97,P ＜ 0.01).The synapse numbers at the middle turn decreased significantly in the 5 W group (F =17.15,P ＜ 0.01),while no significant changes were noted at the apical turn(P ＞0.05).There was no statistical difference in the 2 W group(P ＞0.05).Conclusions The superpulsed CO2 laser-assisted cochleostomy with high-power is accompanied by synaptic injury,while no obvious effects by the low-power laser surgery,which might be the safe strategy to preform cochleostomy.

OBJECTIVETo observe the effects of different concentrations of PPAR gamma agonist rosiglitazone on hypoxia/reoxygenation-induced oxidative stress, cell viability and apoptosis in rat cardiac myocytes.

METHODSCultured rat cardiac myocytes were divided into 5 groups, namely group I (normal group), group II (20 micromo/L ROS group), group III (I/R group), group IV (I/R+20 micromo/L ROS group), and group V (I/R+80 micromo/L ROS group). Group IV and group V were treated with rosiglitazone 12 h before hypoxia/reoxygenation. The changes in cell morphology were observed under optical and transmission electron microscopy, and levels of malondialdehyde (MDA), superoxide dismutase (SOD) activity, and lactate dehydrogenase (LDH) content were determined after the treatment. MTT assay was performed to assess the cell viability and flow cytometry was used to analyze the cell apoptosis.

RESULTSHypoxia/reoxygenation resulted in significantly increased MDA and LDH contents and apoptosis of the cardiac myocytes (P<0.05), but lowered SOD activity and the cell viability (P<0.05). The MDA and LDH contents and apoptotic rate were significantly lower but SOD content and cell vitality significantly higher in groups IV and V than in group III (P<0.05). Group V showed significantly lower MDA and LDH contents and apoptotic rate but higher but SOD content and cell vitality than group IV (P<0.05). Electron microscopy revealed obvious apoptotic changes in group III, and only mild changes were found in group V.

CONCLUSIONRosiglitazone can significantly reduce hypoxia/reoxygenation-induced oxidative stress in cardiac myocytes, improve the cell viability and dose-dependently reduce the apoptotic rate of the cardiac myocytes.

Animals ; Apoptosis ; drug effects ; Cell Hypoxia ; Cell Survival ; drug effects ; Immunohistochemistry ; L-Lactate Dehydrogenase ; metabolism ; Malondialdehyde ; metabolism ; Microscopy, Electron, Transmission ; Myocytes, Cardiac ; cytology ; drug effects ; metabolism ; ultrastructure ; Oxidative Stress ; drug effects ; Oxygen ; metabolism ; PPAR gamma ; agonists ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Thiazolidinediones ; pharmacology

OBJECTIVETo investigate the curative effect and mechanism of transcutaneous laser disc decompression for the treatment of cervical vertigo.

METHODSFrom October 2000 to October 2004, 42 patients with cervical vertigo were treated with percutaneous laser disc decompression by applying a Nd: YAG laser (wavelength is 1064 nm). The postoperative follow-up period was more than 2 months, the change of patients' vertigo were observed.

RESULTSAll the patients were followed up. The mean follow-up period was 7.5 months (from 2 to 36 months). After 2 months of postoperative, 28 patients' vertigo disappeared (67%), 6 patients' vertigo improved (14%), 8 patients' vertigo did not improve. The effective rate was 81%, there was no complication (infection and nerve injury).

CONCLUSIONSCervical intervertebral disc protrusion and cervical spine instability irrigate the neck sympathetic nerve, result in the spasm of vertebral artery, which is the main cause of cervical vertigo. Percutaneous laser disc decompression can decrease intradiscal pressure, increase local temperature, remove the spasm of the vertebral artery. The therapeutic effect for the treatment of cervical vertigo was remarkable.

Adult ; Cervical Vertebrae ; surgery ; Decompression, Surgical ; methods ; Diskectomy, Percutaneous ; methods ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc ; surgery ; Intervertebral Disc Displacement ; complications ; surgery ; Laser Coagulation ; methods ; Male ; Middle Aged ; Treatment Outcome ; Vertigo ; etiology ; surgery

OBJECTIVETo review the treatment of repair gastroschisis with autogenous umbilical cord in 13-year and evaluate its effect.

METHODSTwenty-two newborns who underwent the repair gastroschisis with autogenous umbilical cord between 1992 and 2005. The physical growth, intelligence measuring, area of operation in abdomen in the survived 18 cases were observed and followed-up.

RESULTSEighteen patients recovered uneventfully, survival rate is 82%, their growth is well. They all developed incisional hernia near the operation, 9 cases recovered himself, 2 cases was operated to repair the abdominal hernia, 7 cases is under observed.

CONCLUSIONSThe material is adopted easily in the operating, autogenous umbilical cord is elastic tissue and no toxicity could relax the abdominal press effectively after the operation, the survival rate is high.

Female ; Follow-Up Studies ; Gastroschisis ; surgery ; Hernia, Ventral ; etiology ; Humans ; Infant, Newborn ; Male ; Postoperative Complications ; Umbilical Cord ; transplantation