Aim To investigate the inhibitory effect of paeonol (Pae) on the human esophageal cancer cell line Eca-109 in vitro and in vivo and its effect on apoptosis.Methods Cytotoxic effect of Pae on Eca-109 cells cultured in vitro was measured by MTT assay.Anti-tumor activity was performed on BALB/c nude mice xenografts model.The morphologic changes of tumor tissue were observed under light microscope and transmission electron microscope.The apoptosis index was assessed by TUNEL.Results Pae had significant in- hibitory effect on the proliferation of Eca-109 cells,and the IC50 value was 0.342 mmol?L-1.In the model of human esophageal cancer xenografts in BALB/c nude mice,the inhibitory rates of Pae group (25、50、100、200 mg?kg-1) were 10.67%、23.54%、27.91% and 34.46% respectively.In vivo administration of Pae 100 mg?kg-1 combined with cisplatin 5 mg?kg-1 resulted in a significant inhibition of Eca-109 tumor growth with the inhibitory rate of 77.91%,compared with cisplatin used alone (58.71%).The more apoptotic tumor cells could be seen under light microscope in every theraperutic groups than those in control.Changes of ultrastructure of tumor cells including concentration and side accumulation of the nuclear chromatin,and the fragmentation of the nuclear was observed under transmission electronic microscope.Apoptosis body was also found.The apoptosis indexes of every theraperutic groups were significantly different from the control.Conclusion Pae can inhibit the cell growth and induce apoptosis in human esophageal cancer cell line Eca-109 in vitro and in vivo.

Objective To study the effects of permissive hypercapnia (PHC) on inflammatory factors of pulmonary tuberculosis patients with one-lung ventilation (OLV).Methods Sixty pulmonary tuberculosis patients with selective thoracic operation who were hospitalized at Hangzhou Red-cross Hospital from October 2010 to October 2013 were randomized into three groups by means of random digits table.Twenty patients were enrolled as control group whose arterial partial pressure of carbon dioxide (PaCO2) was maintained at 35-45 mmHg (1 mmHg=0.133 kPa),20 patients as lower PHC group whose PaCO2 were maintained at 50-55 mmHg,and 20 patients as higher PHC group whose PaCO2 were maintained at 56-60 mmHg.The concentrations of serum interleukin (IL)-6 and IL-8 were measured at the time when the patients were in supine position by two-lung ventilation after induction of anesthesia (T1),15 minutes (T2) and 30 minutes after OLV (T3) in lateral position,two hours (T4) and 6 hours after operation (T5).Quantitative data were analyzed by t-test.Results IL-6 at T3 time point in the three groups increased significantly compared with T1 time point (control group:[4.94± 1.60] pg/mL vs [3.32±1.34] pg/mL,t=3.47,P＜0.01; lower PHC group:[5.38±1.42] pg/mL vs [3.69±1.35] pg/mL,t=3.86,P＜0.01; higher PHC group:[5.57±± 1.34] pg/mL vs [3.50 ± 1.22] pg/mL,t=5.11,P＜0.01).IL-8 at T4 time point increased significantly compared with T1 time point (control group:[342.09±56.12] pg/mL vs [194.58 ± 30.07] pg/mL,t =10.36,P＜0.01; lower PHC group:[349.48±43.49] pg/mL vs [193.16±26.27] pg/mL,t=13.80,P＜0.01; higher PHC group:[351.55±38.13] pg/mL vs [186.02±21.08] pg/mL,t=16.99,P＜0.01).IL-6 at the T4 and T5 time points in both lower PHC group and higher PHC group decreased significantly compared with control groupatthe same time points (T4..[38.49± 9.70] and [43.41± 9.01] pg/mL vs [62.35±7.83] pg/mL,t=8.56 and 7.10,both P＜0.01; T5:[56.39±7.47] and [54.77±7.12] pg/mL vs [107.32± 13.97] pg/mL,t=-14.38 and-14.99,both P＜0.01].Conclusions The serum IL-6 and IL-8 levels in pulmonary tuberculosis patients with OLV increase after selective thoracic operation.PHC through low tidal volume could decrease the release of IL-6 and attenuate the pulmonary inflammatory injury in the tuberculosis patients with operation.

0.05). After dividing the patients into early-onset and late-onset subgroups, there were significant differences of DRD4 genotype and allele frequency between early-onset patients and controls (P0.05). Conclusion The results suggested that the polymorphism of DRD4 receptor gene may be associated with early-onset OCD. The 3/4 genetype may be the risk factor of early-onset OCD. Early-onset and late-onset OCD may have different etiology.

Objective To investigate the efficacy of using a calibrator with values assigned with the reference method for improving the comparability of serum gamma-glutamyltransferase (GGT) measurements.Methods The IFCC reference method for GGT was established and the performance was verified by testing a certified reference material (CRM).A calibrator was prepared and its value for GGT was assigned with the reference method.Forty serum samples were measured on different (including HITACHI 7600,7060,7170,7180 and BECKMAN LX20,OLYMPUS AU 400) chemistry analyzers with Zhongsheng GGT reagent kits calibrated with the calibrator.The samples were also measured on the same analyzers using a theoretical factor.Biases of results obtained with the calibration and with the theoretical factor based calculation were compared.Results The reference method resuhs on the CRM agreed the certified value within the stated uncertainty.Serum results calculated from the theoretical factor showed various biases and inter-analyzer variations.When the analyzers were calibrated with the calibrator,the number of results with biases less than 10% became significantly higher and those with biases more than 20% significantly lower.The variation of the results on 5 serum samples was reduced from 11.0%～14.0% to less than 5% by using the calibrator.Conclusion Accuracy and comparability of GGT measurements with of ZhongSheng GGT kits can be improved by using a calibrator that has a value assigned with the reference method.

OBJECTIVETo describe the characteristic of tongue images of patients with lung cancer of different Chinese medicine (CM) syndromes and to reveal the elemental rule on the changes of the tongue images.

METHODSA total of 207 patients with lung cancer were divided into four syndrome groups according to the theory of CM: Fei and Shen deficiency syndrome (Group A, 72 cases), Pi deficiency and phlegm deficiency and phlegm) and Shen (deficiency and phlegm) deficiency syndrome (Group A, 72 cases), Pi (deficiency and phlegm) deficiency and phlegm dampness syndrome (Group B, 57 cases), phlegm-heat retention in Fei (Group C, 36 cases) and yin asthenia generating intrinsic heat syndrome (Group D, 42 cases). The tongue parameters were detected by tongue image digital analysis instrument, and the tongue images were described with qualitative, tongue color and quantitative analysis, respectively. The International Commission on Illumination (CIE) L·a·b (CIELAB) color model was used for the quantitative classification.

RESULTSThere was a significant statistical difference between different syndrome groups of lung cancer on tongue color, coating color, and thickness of tongue coating (P<0.01), and there was significant statistical difference between the four syndrome groups on Lab values of the tongue and coating (P<0.05). The correct identification rate of discriminant function on the raw data was 65.7%, including 72.2% for Group C, 69.4% for Group A, 69.0% for Group D and 54.4% for Group B.

CONCLUSIONSA tongue image digital analysis instrument can objectively describe the tongue features of patients with different syndromes of lung cancer. The tongue diagnosis is very important to syndrome differentiation in CM. Tongue diagnosis should be combined with some important characteristics of syndromes in the future to establish a "combination of four examination methods, including inspection, auscultation, interrogation, and pulse-feeling and palpation" in the tongue diagnostic system.

Color ; Discriminant Analysis ; Humans ; Lung Neoplasms ; pathology ; Medicine, Chinese Traditional ; Tongue

OBJECTIVETo construct a subtractive cDNA library of genes transactivated by PS1TP1 protein with suppression subtractive hybridization (SSH) technique.

METHODSSuppression subtractive hybridization technique and bioinformatics technique were used, the mRNA from HepG2 cells transfected with pcDNA3.1 (-) -PS1TP1 and pcDNA3. 1 (-) empty vector was isolated, respectively; cDNA underwent two times of nested PCR, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library.

RESULTSThe subtractive library of genes transactivated by PS1TP1 was constructed successfully. Sequence analysis was performed in 43 clones randomly, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 12 coding sequences were gotten, which consisted of 10 known and 2 unknown ones.

CONCLUSIONThe obtained sequences may be target genes transactivated by PS1TP1 protein among which some genes coding proteins involved in cell cycle regulation, metabolism, immunity and cell apoptosis. This finding brought some clues for studying the biological functions of PS1T1.

Hepatitis B ; genetics ; Humans ; Nucleic Acid Hybridization ; methods ; Transcriptional Activation ; genetics ; Viral Proteins ; genetics

To clarify the important functional residues in the active site of N-myristoyltransferase (NMT), a novel antifungal drug target, and to guide the design of specific inhibitors, multiple sequence alignments were performed on the NMT family and thus evolutionary trace was constructed. The important functional residues in myristoyl CoA binding site, catalytic center and inhibitor binding site of NMT family were identified by ET analysis. The trace residues were mapped onto the active site of CaNMT. Trpl26, Asn175 and Thr211 are highly conserved trace residues and do not interact with current NMT inhibitors, which are potential novel drug binding sites for the novel inhibitor design. Pro338, Leu350, Ile352 and Ala353 are class-specific trace residues, which are important for the optimization of current NMT inhibitors. The trace residues identified by ET analysis are of great importance to study the structure-function relationship and also to guide the design of specific inhibitors.
Acyl Coenzyme A ; metabolism ; Acyltransferases ; chemistry ; genetics ; metabolism ; Amino Acid Sequence ; Animals ; Binding Sites ; Conserved Sequence ; Enzyme Inhibitors ; chemistry ; pharmacology ; Evolution, Molecular ; Humans ; Imidazoles ; chemistry ; pharmacology ; Models, Molecular ; Molecular Sequence Data ; Oligopeptides ; chemistry ; pharmacology ; Phylogeny ; Protein Structure, Tertiary ; Sequence Homology, Amino Acid

Objective To retrospectively analyze treatment status quo of giant cell tumor of bone around the knee in several institutes,and to investigate risk factors affecting selection of surgical manners,tumor recurrence,and functional outcomes.Methods A total of 222 patients with giant cell tumor of bone around the knee confirmed by pathology,who had undergone surgical treatment in 5 institutes from March 2000 to May 2012,were enrolled in this study.There were 120 males and 102 females,with an average age of 35.5 years.The epidemiology,clinical and radiographic features and risk factors affecting selection of surgical manners,tumor recurrence,and functional outcomes were analyzed.Results Intralesional curettage was performed in 128 patients(57.6％),intralesional curettage combined with partial resection in 13 patients (5.8％),and marginal excision in 79 patients (35.5％).A total of 159 patients were followed up.The local recurrence rate was 19.0％ for patients treated with intralesional curettage,8.9％ for marginal excision and 0％ for intralesional curettage combined with partial resection,and the total local recurrence rate was 14.5％(23/159).The mean duration from primary operation to recurrence was 23.9 months.Univariate analysis indicated that surgical manner was the only factor affecting local recurrence.Pathological fracture,Campanacci grades,Enneking system as well as soft tissue mass had a significant influence on the selection of surgical manners.The mean MSTS score after intralesional curettage was obviously higher than that after marginal excision.Conclusion The surgical manner is the only factor affecting local recurrence and limb function.

OBJECTIVETo investigate the antimicrobial resistance and penicillin resistance-associated genes (TEM and pbp2B) of Streptococcus pneumoniae (S. pneumoniae) isolated from sputum specimens of Guangzhou children with respiratory tract infection.

METHODSE-test and Kirby-Bauer methods were applied to detect the antibiotic susceptibility of 44 strains of S. pneumoniae. PCR was used to detect resistance genes pbp2B and TEM, followed by DNA sequence analysis of pbp2B gene. The sequence results were compared to those of penicillin-susceptible S. pneumoniae R6.

RESULTSOf the 44 isolates of S. pneumoniae, only 5 (11.4%) were susceptible to penicillin. All strains were resistant to erythromycin but susceptible to ofloxacin and vancomycin. The resistance rate of the isolates to clindamycin and trimoxazole was more than 90%. The S. pneumoniae isolates showed a high susceptibility to amoxicillin, imipenem and ceftriaxone, with a resistance rate of 0, 2.6% and 3.9%, respectively. The sequence analysis showed that more than 99% nucleotide sequence of pbp2B gene of five penicillin-susceptible isolates was the same as penicillin-susceptible S. pneumoniae R6, without any amino acid replacement. Site mutation was found in the remaining 39 penicillin-nonsusceptible isolates with a nucleotide mutation rate ranging from 13.2% to 23.1% and amino acid replacement rate from 6.5% to 10.9%. The 39 penicillin-nonsusceptible isolates were classified into 4 types according to the mutation site between Ser391 and Thr492 of pbp2B: type I (n=30), type II (n=7), type III (n=1) and type IV (n=1). No TEM gene was detected in all the 44 S. pneumoniae isolates.

CONCLUSIONSThe S.pneumoniae isolates from Guangzhou children with respiratory tract infection are resistant to penicillin and erythromycin. Amoxicillin and the third generation cephalosporin may be recommended for treating S. pneumoniae infection. The mutation of pbp2B gene plays an important role in the development of S. pneumoniae resistance to penicillin.

Aminoacyltransferases ; genetics ; Child, Preschool ; Drug Resistance, Bacterial ; genetics ; Female ; Humans ; Infant ; Male ; Microbial Sensitivity Tests ; Penicillin Resistance ; genetics ; Penicillin-Binding Proteins ; genetics ; Respiratory Tract Infections ; microbiology ; Streptococcus pneumoniae ; drug effects ; genetics ; beta-Lactamases ; genetics

Objective To investigate the accuracy and comparability of ?-glutamyltransferase (?- GT) measurement results on human serum samples and controI materials before and after calibration with a common human serum calibrator.Methods A human serum calibrator was prepared by pooling fresh serum aliquots and assigning a value for ?-GT with the IFCC reference method.The calibrator together with 5 human serum samples and 10 control samples were sent to 15 clinical laboratories and the sermn and control samples were measured with different analytical systems before and after a calibration with the calibrator.The results were analyzed for biases and interlaboratory variations.Results For the serum samples,the calibration resulted in reductions in biases from -9.0%～-14.2% to -0.8%～-7.9%,and in interlaboratory variations from 6.9%～11.6% to 2.8%～4.4%.No improvement was observed on the control samples.Conclusions Accuracy and comparability of serum ?-GT measurements can be improved by using a common human serum calibrator.Some control materials may not be commutable for human serum in ?-GT measurements.