OBJECTIVETo establish a full-quantified fingerprint method for analyzing Chinese herbal additives by HPLC.

METHODA HPLC in combination with PDA detector was applied with a phenomenex luna C18 (4.6 mm x 250 mm, 5 microm) column by gradient elution using acetonitrile-0.1% formic acid solution as the mobile phase. The flow rate was 1 mL x min(-1) and the detection wavelength was set at full spectrum scan.

RESULTAccording to the selected chromatographic conditions, the full-quantified fingerprint of the Chinese herbal additive has good precision, reproducibility, stability and recovery.

CONCLUSIONThe HPLC method developed for simultaneous determination of seven compounds is simple and valid. It can be used for quality evaluation and quality control of Chinese herbal additive and its processing products.

Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis

OBJECTIVETo explore the causes of nonspecific chronic cough in children and relationship between transient receptor potential vanilloid 1 (TRPV1) gene polymorphisms and nonspecific chronic cough.

METHODSA total of 195 children with chronic cough were followed up half a month, one month and three months after their first visit to hospital. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to examine polymorphisms of the TRPV1 gene in the children. A total of 205 healthy or surgical children without chronic cough served as the control group.

RESULTSThe etiologic distribution of the 195 children with chronic cough was as follows: 96 (49.2%) cases of cough variant asthma (CVA), 48 (24.6%) cases of CVA complicated by upper airway cough syndrome (UACS), 34 (17.4%) cases of post-infectious cough, and 17 (8.7%) cases of UACS. Three genotypes were identified in both groups at positions rs222747 (CC, GC and GG), rs222748 (CC, TC and TT) and rs8065080 (CC, TC and TT). The frequencies of genotype and allele at position rs222747 did not accord with the law of Hardy-Weinberg. There was no significant difference in frequencies of genotype and allele at positions rs222748 and rs8065080 between the two groups.

CONCLUSIONSCVA, UACS and post-infectious cough are common causes of nonspecific chronic cough in children. TRPV1 gene polymorphisms at positions rs222748 and rs8065080 may be unrelated to nonspecific chronic cough in children.

Adolescent ; Alleles ; Child ; Child, Preschool ; Chronic Disease ; Cough ; etiology ; genetics ; Female ; Genotype ; Humans ; Infant ; Male ; Polymorphism, Genetic ; TRPV Cation Channels ; genetics

OBJECTIVETo investigate the proportion of incidence of children with non-specific chronic cough in Chongqing and analyze the characteristics of etiology during the follow-up.

METHODDiagnostic criteria were defined for children with non-specific chronic cough according to the Guidelines of diagnosis and therapy for children with chronic cough that were formulated by the Subspecialty Group, Society of Pediatrics, Chinese Medical Association and Chinese Journal of Pediatrics in 2008. Totally 266 patients in whom cough was the main or the only symptom,lasting > 4 weeks, presenting to Asthma Center of Children's Hospital, Chongqing Medical University between June 2008 and April 2009 were recruited into this study. Based on the Guidelines, diagnosis was made after taking history, physical examination and assistant examination. After etiological treatment, the patients were followed up during the second week, the fourth week and the twelfth week. Etiological diagnosis was confirmed if cough was resolved after specific therapy. If cough was not resolved,the diagnosis was rechecked and a new therapy was applied.

RESULTTotally 125 (47.0%) patients received final diagnoses of cough variant asthma (CVA), 58 (21.8%) was CVA and upper airway cough syndrome (UACS), 44 (16.5%) was diagnosed postinfection cough, 35 (13.2%) of UACS. In different age groups, the proportion of incidence of etiological agents is statistically distinct. In the ≤ 3 years old group, 35 patients (70.0%) were diagnosed CVA, 10 (20.0%) was postinfection cough; in 3 - 6 years group, 71 patients (50.7%) had CVA; the incidence of UACS was significantly higher in ≥ 6 years group.

CONCLUSIONIt is concluded that CVA, CVA and UACS, post infection cough, and simple UACS were identified as the three top reasons for children with chronic cough in Chongqing. Children with chronic cough of different age groups had different etiology of cough. The characteristic of each etiology need further study.

Adolescent ; Asthma ; epidemiology ; Child ; Child, Preschool ; China ; epidemiology ; Chronic Disease ; Cough ; epidemiology ; etiology ; microbiology ; Follow-Up Studies ; Humans ; Incidence ; Infant ; Infection ; epidemiology

OBJECTIVETo study the value of 3D visualization technique in breast-preserving surgery for breast cancer with immediate breast reconstruction using laparoscopically harvested pedicled latissimus dorsi muscle flap.

METHODSFrom January, 2015 to May, 2016, 30 patients with breast cancer underwent breast-preserving surgery with immediate breast reconstruction using pedicled latissimus dorsi muscle flap. The CT data of the arterial phase and venous phase were collected preoperatively and imported into the self-developed medical image 3D visualization system for image segmentation and 3D reconstruction. The 3D models were imported into the simulation surgery platform for virtual surgery to prepare for subsequent surgeries. The cosmetic outcomes of the patients were evaluated 6 months after the surgery. Another 18 patients with breast cancer who underwent laparoscopic latissimus dorsi muscle breast reconstruction without using 3D visualization technique from January to December, 2014 served as the control group. The data of the operative time, intraoperative blood loss and postoperative appearance of the breasts were analyzed.

RESULTSThe reconstructed 3D model clearly displayed the anatomical structures of the breast, armpit, latissimus dorsi muscle and vessels and their anatomical relationship in all the 30 cases. Immediate breast reconstruction was performed successfully in all the cases with median operation time of 226 min (range, 210 to 420 min), a median blood loss of 95 mL (range, 73 to 132 mL). Evaluation of the appearance of the breast showed excellent results in 22 cases, good appearance in 6 cases and acceptable appearance in 2 cases. In the control group, the median operation time was 283 min (range, 256 to 313 min) and the median blood loss was 107 mL (range, 79 to 147 mL) with excellent appearance of the breasts in 10 cases, good appearance in 4 cases and acceptable appearance in 4 cases.

CONCLUSION3D reconstruction technique can clearly display the morphology of the latissimus dorsi and the thoracic dorsal artery, allows calculation of the volume of the breast and the latissimus dorsi, and helps in defining the scope of resection of the latissimus dorsi to avoid injuries of the pedicled vessels. This technique also helps to shorten the operation time, reduce intraoperative bleeding, and improve the appearance of the reconstructed breast using pedicled latissimus dorsi muscle flap.

OBJECTIVETo explore the impact of Foxp3 expression and CD(4)(+)CD(25)(+) regulatory T cells on pathogenesis of childhood asthma.

METHODSTotally 15 patients with acute asthma exacerbation, 15 children with asthma remission and 10 children who were hospitalized for skeleton deformity without atopic disorders or history of allergic diseases or respiratory infections within a month as controls were recruited in this study from Sep. 2004 to Mar. 2005. The percentage of CD(4)(+)CD(25)(+) T cells were detected by 2-color flow cytometry. The levels of interleukin (IL)-4, IL-10, interferon (IFN)-gamma, transforming growth factor (TGF)-beta in plasma and supernatant were assayed by ELISA. Both the asthmatic children and the control children were selected to induce sputum by hypertonic saline. Sputum was processed for detecting the expression of Foxp3-mRNA. The expression of Foxp3-mRNA in both sputum and PBMC was detected by semi-quantitative RT-PCR with beta-actin as internal control.

RESULTSThe percentage of CD(4)(+)CD(25)(+) regulatory T cells in exacerbation and remission asthmatic children was significantly lower than that of the control children both prestimulation [(10.1 +/- 2.1)% vs. (15.5 +/- 2.7)%, (11.7 +/- 2.5)% vs. (15.5 +/- 2.7)%, P < 0.05] and poststimulation with PHA [(12.4 +/- 2.3)% vs. (26.9 +/- 3.8)%, (17.3 +/- 3.2)% vs. (26.9 +/- 3.8)%, P < 0.05]. The percentage of CD(4)(+)CD(25)(+) regulatory T cells was significantly higher after PHA stimulation in normal children [(15.5 +/- 2.7)% vs. (26.9 +/- 3.8)%, P < 0.01]. The expression of Foxp3-mRNA (Foxp3/beta-actin) in asthmatic children was significantly lower than that in the control children in both PBMC and induced sputum. The expression of Foxp3-mRNA in PBMC was significantly higher after PHA stimulation in the control children (0.77 +/- 0.22 vs. 1.07 +/- 0.21, P < 0.05). However, there was no significant difference in Foxp3-mRNA expression in asthmatic children pre and post PHA stimulation. A significant positive correlation between the Foxp3-mRNA expression and the percentage of CD(4)(+)CD(25)(+) regulatory T cells was detected. The levels of IFN-gamma and TGF-beta were significantly lower in asthmatic children than those in the control children, and the levels of IFN-gamma and TGF-beta correlated positively with Foxp3-mRNA expression and the percentage of CD(4)(+)CD(25)(+) regulatory T cells. The level of IL-4 both in plasma and supernatant was higher in asthmatic children. The levels of IL-10 was higher only in exacerbation than in control children, the levels of IL-4 and IL-10 had no correlation with Foxp3-mRNA expression and the percentage of CD(4)(+)CD(25)(+) regulatory T cells.

CONCLUSIONInsufficient secretion of TGF-beta, decreased Foxp3 expression, insufficient number of CD(4)(+)CD(25)(+) regulatory T cells and the defective ability of converting CD(4)(+)CD(25)(-) T cells to CD(4)(+)CD(25)(+) regulatory T cells might play an important role in pathogenesis of asthma.

Asthma ; physiopathology ; Case-Control Studies ; Child ; Cytokines ; analysis ; blood ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Forkhead Transcription Factors ; genetics ; metabolism ; Humans ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sputum ; chemistry ; T-Lymphocytes, Regulatory ; physiology

OBJECTIVETo investigate the relationship between fetal chromosomal karyotype and early spontaneous abortion, and the effect of the environmental factors on spontaneous abortion.

METHODSChoronic villi from 252 cases of missed abortion were sampled as patient group and 50 normal pregnancies as control group. Chorionic villi were cultured and karyotype analysis was performed by G-banding. Clinical information was collected.

RESULTSThe rate of chorion chromosome abnormality in the patient group was 58.09%, significantly higher than that in the control group (4.17%) (P<0.01). Among the 140 cases of karyotype abnormalities, 81 were trisomy, 29 were monosomy X and 17 were polyploidy, accounting for 57.86%, 20.71% and 12.14% of total abnormalities, respectively. Long time and low dose radiation exposure of the pregnant female seemed to be related with spontaneous abortion(P<0.01).

CONCLUSIONChorion chromosome abnormality is a major reason of early spontaneous abortion and karyotype analysis of chorionic villus is of clinical importance. For pregnant women, long-term exposure to computers and television seems a risk factor for missed abortion.

Abortion, Missed ; genetics ; Adolescent ; Adult ; Case-Control Studies ; Chorionic Villi ; metabolism ; Chromosome Aberrations ; Female ; Humans ; Karyotyping ; Middle Aged ; Pregnancy ; Risk Factors ; Young Adult

OBJECTIVETo examine vibrio cholera (V.C) in aquatic products of littoral area, Zhejiang Province and to provide scientific evidence for administration of aquatic products and cholera epidemic control.

METHODSAll 990 samples of aquatic products collected from local markets, eateries and aquafarms in three chosen areas. Samples were proliferated in alkaline liquid medium, and purified in NO: 4 medium, the isolations were identified biochemically, and phenotype of strains were defined by phagocyte and coagulation with V.C. diagnostic serum. Three virulence genes (ctx, ace, zct) of the isolated strains were detected by polymerase chain reaction (PCR).

RESULTSThere were 1.41% samples caught by V.C., having a carrying rate highest in turtles of 8.9%. 14 strains were defined as three serogroups, and the numbers of Inaba, Ogawa, and Hikojima types were 2, 2, 10 respectively. Virulence genes had detected in 9 of 12 stains. All genes were detected in 5 strains, only ZOT genes in 3 strains, and both CTX and ACE genes in 1 strain.

CONCLUSIONSAquatic products from inshore in Zhejiang Province caught with V.C. strains might be divided into three serogroups. Most of them should be virulence genes. Cholera epidemic outbreak might be caused by those contaminated products.

China ; Food Microbiology ; Genes, Bacterial ; Seafood ; microbiology ; Vibrio cholerae ; genetics ; isolation & purification ; Virulence Factors ; genetics

Aim To discuss the neuroprotection of an-tioxidant α-phenyl-N-tert-butyl nitrone ( PBN) in Par-kinson's disease ( PD) model. Methods The neuro-protective effects of PBN was explored on 1-methyl-4-phenylpyridinium/1-methyl-4-phenyl-1,2,3,6-tetrahy-dropyridine ( MPP+/MPTP ) induced PD models through a chemical releasing of free radicals in neurons and in vivo. Results PBN significantly scavenged chemical derived free radicals, reduced MPP+induced SH-SY5Y injury and enhanced neurons' viability. In MPTP induced PD model, PBN significantly enhanced the number of tyrosine hydroxylase ( TH)-positive do-pamine ( DA) neurons in the substantia nigra, restored the expression of Nrf2 and HO-1, and raised striatal contents of DA and its metabolites. Conclusion PBN has a potent neuroprotective effect against MPP+/MPTP induced PD models.

OBJECTIVETo investigate the role of α-enolase (ENO1) in regulating glucose metabolism and cell growth in human glioma cells.

METHODSGlucose uptake and lactate generation were assessed to evaluate the changes in glucose metabolism in human glioma U251 cells with small interfering RNA (siRNA)-mediated ENO1 knockdown. MTT assay and 5-ethynyl-2'-deoxyuridine (EdU) staining were used to examine the cell growth and cell cycle changes following siRNA transfection of the cells.

RESULTSTransfection of U251 cells with siRNA-ENO1 markedly reduced glucose uptake (P=0.023) and lactate generation (P=0.007) in the cells and resulted in significant suppression of cell proliferation (*P<0.05) since the second day following the transfection. Transfection with siRNA-ENO1 also obviously suppressed cell cycle G1/S transition in the cells (P=0.0425). The expressions of HK2 and LDHA, the marker genes for glucose metabolism, were significantly down-regulated in the cells with siRNA-mediated ENO1 knockdown.

CONCLUSIONENO1 as a potential oncogene promotes glioma cell growth by positively modulating glucose metabolism.

BACKGROUNDIncomplete right bundle branch block (ICRBBB) is commonly associated with atrial septal defect (ASD), but lacks sufficient diagnostic test characteristics. An abnormal T wave is also often observed in ASD, with horizontal or inverted displacement of the proximal T wave limb in the right precordial leads, termed "defective T wave" (DTW).

METHODSWe examined the diagnostic test characteristics of combining ICRBBB with DTW as a new index to diagnose ASD. A total of 132 consecutive patients with ASD and 132 cases of age/gender-matched controls without ASD were enrolled.

RESULTSSensitivities of DTW, ICRBBB, and both were 87.1% - 87.9%. Specificities were 97.0%, 96.2%, and 100%, respectively. Positive predictive values were 1.3%, 1.1%, and 100.0% respectively, while negative predictive values were 99.9% for each.

CONCLUSIONCombining ICRBBB with DTW in electrocardiogram (ECG) as a new index significantly increased the specificity and positive predictive values while maintaining a high sensitivity in diagnosing ASD.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bundle-Branch Block ; diagnosis ; physiopathology ; Child ; Child, Preschool ; Electrocardiography ; Female ; Heart Septal Defects, Atrial ; diagnosis ; Humans ; Male ; Middle Aged ; Predictive Value of Tests ; Sensitivity and Specificity