To study the toxicokinetics of bakuchiol, hepatic and renal toxicity in rats after single oral administration of Psoraleae Fructus and combined with Glycyrrhizae Radix et Rhizoma, in order to provide scientific evidences for clinical safe medication use. A total of 35 SD rats were randomly divided into seven groups: vehicle (distilled water) control group, Glycyrrhizae Radix et Rhizoma group, positive control (aristolochic acid A) group, Psoraleae Fructus (40 g x kg(-1)) group( both male and female rats), Psoraleae Fructus and Glycyrrhizae Radix et Rhizoma (40 +20) g x kg(-1) group (both male and female rats). HPLC-UV method was used to determine the concentration of bakuchiol in rat plasma at different time points after single oral administration. Plasma alanine transaminase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), plasma creatinine (Cr), N-acetyl-β-D-glucosaminidase (NAG) and kidney injury molecule 1 (Kim-1) were measured after administration for 24 h. The main toxicokinetics parameters of bakuchiol in rats exert significantly gender difference. When Psoraleae Fructus combination with Glycyrrhizae Radix et Rhizoma, the total area under the plasma concentration-time curve( AUC), C(max), and plasma clearance (CL) of bakuchiol were increased, respectively; CL, half-life (t½) were decreased, and T(max) were prolonged. The biochemical indicators (including ALT, AST, BUN, Cr and KIM-1 level) in different dose of Psoraleae Fructus groups, were found no statistically significant difference when compared with vehicle control group. The level of NAG in both Psoraleae Fructus and compatibility with Glycyrrhizae Radix et Rhizoma groups were significant increased (P < 0.05). There are obvious effects on toxicokinetics of bakuchiol in rats when Psoraleae Fructus combined with Glycyrrhizae Radix et Rhizoma. Renal toxicity induced by Psoraleae Fructus at high dose was observed after single oral administration and no liver damage in rats was found.
Administration, Oral ; Animals ; Female ; Glycyrrhiza ; toxicity ; Kidney ; drug effects ; Liver ; drug effects ; Male ; Phenols ; pharmacokinetics ; toxicity ; Psoralea ; toxicity ; Rats ; Rats, Sprague-Dawley ; Rhizome ; toxicity ; Toxicokinetics

An antibiotic-producing bacterium, which was numbered as 20 #-5, was separated from the soil in Chongqing. It was identified as the member of pseudomonas. Gram positive bacteria are badly suppressed by it. The antibiotic secreted by 20 #-5 can endure 100℃ for half an hour, and it can also go through the ultrafiltration membrane with pores of 0.22?m.

OBJECTIVETo compare the differences in the clinical function and lumbar and abdominal myodynamia in patiants of lumbar disc herniation treated with moxibustion at Dazhui (GV 14) and Guanyuan (CV 4) and acupuncture.

METHODSForty cases were randomized into a moxibustion group and an acupuncture group, 20 cases in each group. In the moxibustion group, the warm moxibustion was applied alternatively at Dazhui (GV 14) and Guanyuan (CV 4) once every other day, 1 h each time, once every day. In the acupuncture group, acupuncture was applied to the corresponding acupoints based on the affected lumbar vertebras, such as Jiaji (EX-B 2), Shens-hu (BL 23), Dachangshu (BL 25) and Huantiao (GB 30), etc. , once evey day 30 min each time. The treatment for 3 weeks was taken as one session in each group. Totally, one session treatment was required. Surface electromyography (SEMG) of bilateral paraspinal muscle and rectus muscle, and JOA score of low back pain were observed in the two groups.

RESULTS(1) JOA score: the score of subjective symptoms, score of activity of daily living (ADL) and total score were improved obviously as compared with those before treatment in the two groups (P<0.01, P<0.05). The results of subjective symptoms score, score of ADL and total score in the acupuncture group were superior to those in the moxibustion group after treatment (6.95+/-0.94 vs 5.50 +/-0.89,10. 90+/-1.86 vs 8.90+/- 1. 92,22.50 +/- 2.82 vs 19.35 +/- 2. 70, all P<0. 05). (2) SEMG comparison: root-mean-square value (RMS) was all reduced in SEMG of the anteflexion, rear protraction, orthostatism, bilateral bending and neck and leg rear flexion for strengthening lumbar muscle as compared with those before treatment in the two groups (P< 0.05, P<0. 01). RMS of the anteflexion and bilateral bending in the acupunture group were reduced much obviously as compared with the moxibustion group. In terms of sitting position anteflexion, rear protraction, orthostatism, bilateral bending and neck and leg rear flexion for strengthening lumbar muscle, median frequency (MF) after treatment was all improved as compared with that before treatment in the two groups (P<0. 05, P<0. 01). In terms of anteflexion, the electrode MF after treatment was improved much obviously in the acupuncture group(P<0. 05).

CONCLUSIONMoxibustion at Dazhui (GV 14) and Guanyuan (CV 4) and conventional acupuncture all improve muscle function, relieve muscle fatigue, increase the ability of anti-muscle fatigue, strengthen lumbar vertebral stability, release subjective symptoms and improve ADL. But, the effects of moxibustion are slightly lower than those of acupuncture.

Acupuncture Therapy ; Adult ; Aged ; Electromyography ; Female ; Humans ; Intervertebral Disc Displacement ; physiopathology ; therapy ; Lumbar Vertebrae ; physiopathology ; Male ; Middle Aged ; Moxibustion ; Young Adult

OBJECTIVETo study the loss of heterozygosity (LOH) on chromosome 3p in breast cancers and precancerous lesion.

METHODSLOH at 11 microsatellite loci was detected in 41 cases of breast cancers and 12 cases of precancerous lesion by polymerase chain reaction and silver stain. The expressions of ER, PR, FHIT and hMLH1 were detected in breast cancer by immunohistochemistry.

RESULTSLOH on 3p was detected in 97% of breast cancers. D3S1295, D3S1029 and D3S1038 located at 3p14, 3p21-p22 and 3p25 were identified as the loci with most frequent LOH (53.1%, 43.6% and 52.5%). LOH of D3S1038 and expression of hMLH1 protein correlated with several clinicopathological features. LOH of D3S1295 had significant negative correlation with the expression of FHIT. In breast precancerous lesions, LOH on 3p was detected in 41.7% lesions. D3S1295 and D3S1029 were also identified as the most frequent LOH locus (27.3% and 16.7%). The smallest common LOH region seems likely lie between 3p14 and 3p25.

CONCLUSIONSThe smallest LOH region indicates the existence of breast tumor related genes and some of them affect gene expression.

Acid Anhydride Hydrolases ; metabolism ; Adult ; Aged ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Chromosomes, Human, Pair 3 ; genetics ; Female ; Humans ; Immunohistochemistry ; Loss of Heterozygosity ; Microsatellite Repeats ; genetics ; Middle Aged ; Neoplasm Proteins ; metabolism ; Polymerase Chain Reaction ; Precancerous Conditions ; genetics ; pathology ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism

Objective To establish a UPLC-MS/MS method for determination of 10 anti-rheumatic constituents illegally added in Chinese traditional medicine and health products preparation.Methods The column was ACQUITY UPLC BEHC18 (50 mm× 2.1 mm,1.7 μm),The mobile phase was acetonitrile-Ammonium acetate solution (containing 0.1％ Acetic acid) with gradient elution at a flow rate of 0.3 mL/min.The ion source was electrospray ionization (ESI),Multiple-reaction monitoring (MRM) was performed to identify and quantify 10 anti-rheumatic constituents.Results 10 linear calibration curves were obtained with r ≥ 0.996 1.The recoveries were determinated at three concentration and ranged from 92.5％ to 101.8％.The precision of the method was shown by RSD (n =5) ranged from 0.9％ to 3.1％.The ranges of limit of detection were from 0.001 5 to 0.018 μg,and quantitation were from 0.004 5 to 0.55 μg.The illegally added chemicals were detected with 10 batches of 27 batches of samples.Conclusion The method were simple,sensitivity,accurate,and can be used to detect Anti-rheumatic constituents illegally added in Chinese traditional medicine and health products.

Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3G (APOBEC3G) cDNA was amplified from total RNA prepared from nonpermissive H9 cells by RT-PCR. APOBEC3G cDNA is 1155nt long, encoding 384 amino acids. The APOBEC3G gene was then cloned into the eukaryotic expression vector pEGFP-C3. The generated pEGFP-3G construct was then transfected into CD4+ HeLa cell to determine the expression and the subcellular localization of GFP-APOBEC3G fusion protein. Under CLSM the localization of the expressed GFP-APOBEC3G in the cytoplasm of CD4+ HeLa cells was observed.
APOBEC-3G Deaminase ; Cell Line ; Cytidine Deaminase ; Cytoplasm ; metabolism ; DNA, Complementary ; genetics ; isolation & purification ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; HeLa Cells ; Humans ; Microscopy, Confocal ; methods ; Nucleoside Deaminases ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the prevalence of viral infections and putative association of viral infection with illness severity in young children with severe lower respiratory tract infection (LRTI) in Chongqing.

METHODRespiratory secretion specimens were collected from 119 hospitalized patients with severe pneumonia from December 2006 to March 2008.After being processed, the samples were detected for respiratory viruses including respiratory syncytial virus (RSV), adenovirus (ADV), human metapneumovirus (hMPV), human bocavirus (HBoV), parainfluenza virus 1, 2, 3 (PIV 1, 2, 3), influenza virus A and B (IVA and IVB) either by PCR or RT-PCR. Clinical data were analyzed along with virological data by using appropriate statistical methods.

RESULTViral pathogens were identified in specimens of 86 (72.3%) cases, among which RSV was detected in 49 (41.2%) patients. More than one virus was detected in 23 individual (26.7%) samples, of which 19 were dual positive for RSV and another virus. Bacterial cultures were performed for 69 patients. Both bacterial and viral pathogens were identified in 53 (76.8%) patients. Bacterial and viral coinfection was demonstrated in samples from 41 (59.4%) cases.

CONCLUSIONViral pathogens are the main etiology of severe pneumonia in young children in Chongqing area during the study period. RSV was the most frequent viral pathogens, followed by ADV and hMPV. Coinfection with respiratory common viruses was relatively common, though co-infection with viruses did not appear to aggravate the patients' condition.

Adenoviridae ; isolation & purification ; Child, Preschool ; China ; epidemiology ; Human bocavirus ; isolation & purification ; Humans ; Infant ; Infant, Newborn ; Influenza A virus ; isolation & purification ; Metapneumovirus ; isolation & purification ; Pneumonia, Bacterial ; microbiology ; virology ; Pneumonia, Viral ; microbiology ; virology ; Respiratory Syncytial Viruses ; isolation & purification ; Virus Diseases ; virology

OBJECTIVETo investigate the mechanism underlying the curcumin-induced apoptosis of nasopharyngeal carcinoma (NPC) cell line NCE cells.

METHODSThe characteristics of apoptosis were identified by observation acridine orange and ethidium bromide stains, ultrastructure assay, DNA fragmentation assay and TdT-mediated dUTP nick end labeling method (TUNEL). Mitochondrial membrane potential (delta psi m), activity of caspase-3, cytosol cytochrome C and expression of gene Fas were determined by flow cytometry (FCM), Western Blot and reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSSeveral evidences of apoptosis were obtained from curcumin-treated NCE cells by acridine orange and ethidium bromide stains, ultrastructure identification, DNA fragmentation assay and TUNEL staining. And the mean TUNEL-positive rates increased significantly at the 3 different time points (12 h, 24 h and 48 h; 25.6%, 40.3% and 54.5%, respectively). In the curcumin-treated-groups, delta psi m altered significantly and the positive rates increased in a time-dependent manner. At the 3 different time points, the mean positive rates were 26.8%, 42.3% and 68.2%, respectively. When caspase-3 activity was detected, 80.5% cells presented proteases activities after 12 h incubation with curcumin. Western Blot analysis showed that cytoplasmic cytochrome C increased significantly after incubation with curcumin. Flow cytometry and RT-PCR analysis showed that curcumin could up-regulate the Fas expression in time-depended manner , the positive rates of Fas protein increased from 33.6% to 89.9%.

CONCLUSIONSCurcumin induced apoptosis of NCE cells both through mitochondria-dependent pathway and death receptor pathway.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Curcumin ; pharmacology ; Humans ; Membrane Potential, Mitochondrial ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; fas Receptor ; metabolism

OBJECTIVETo developed a multiplex RT-PCR assay for simultaneous screening of type A, B and novel A (H1N1) influenza viruses.

METHODSTwo pairs of universal primers in were designed for amplifying the M gene and NS gene of type A and B influenza viruses, respectively. A pair of specific primers of HA gene was designed to detect novel A (H1N1) influenza virus. A one-step method was used to establish the multiplex RT-PCR system. A blinded experiment was carried out to validate the accuracy of this assay in comparison with the results of real-time fluorescence RT-PCR. The clinical practicability and efficacy of this assay was also evaluated.

RESULTSThe RT-PCR products were analyzed using agarose gel electrophoresis, which yielded distinct bands of the target fragments without non-specific reactions, suggesting the high efficiency and specificity of the multiplex RT-PCR. Blinded study of 50 samples demonstrated a concordance rate of 100%.

CONCLUSIONThis multiplex RT-PCR assay allows one-step simultaneous detection of type A, B and novel A (H1N1) influenza viruses rapidly and accurately, and provides a valuable low-cost screening technique for influenza epidemic monitoring and early diagnosis.

Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza B virus ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Time Factors ; Viral Matrix Proteins ; genetics ; Viral Nonstructural Proteins ; genetics

Objective To evaluate endoscopic ultrasonography (EUS) in diagnosis of gastric submucosal tumors (SMTs), and analyze the characteristics of gastric SMTs under EUS. Methods Clinical data of 614 patients with gastric SMTs, who were evaluated by EUS and underwent endoscopic submucosal dissection (ESD) from September 2008 to December 2016, were retrospectively analyzed. The golden standard for lession origins was the intraoperative diagnosis of ESD, and that for pathological types was the combination of postoperative pathological and immunohistochemical findings. The consistency of diagnosis of EUS was evaluated, and the characteristics of lesions under EUS were analyzed. Results The total consistency in diagnosing lesion origins was 91.25％ between EUS and intraoperative results of ESD, and the consistency in diagnosing lesion originated from the muscularis mucosa, submucosa and muscularis propria was 66.67％, 80.85％ and 94.50％, respectively. The total consistency in pathological types was 65.99％ between EUS and postoperative pathological results, and the consistency of gastrointestinal stromal tumor (GIST), leiomyoma, ectopic pancreas and lipoma was 91.85％, 18.56％, 79.76％ and 90.70％, respectively. Conclusion EUS can initially determine the origins and types of gastric SMTs, providing a more accurate basis for endoscopic treatment, but there were some limitations on the diagnosis of leiomyoma and some rare lesions such as hamartoma, inflammatory fibrous polyps, carcinoid, fibroma, etc. Thus, if necessary, the lesion should be removed so as to make a definite diagnosis and prevent malignant change.