Comparative study on appearance characters and internal structure of cultivated and wild Ganoderma lucidum in Huoshan / 中国中药杂志

Through the comparative study on the appearance characters and internal structure of cultivated and wild Ganoderma lucidum in Huoshan,this paper provides a reference for the further study of G. lucidum. In this study,the similarities and differences between cultivated G. lucidum " Huozhi No. 1" and wild G. lucidum in Huoshan were compared by means of character observation,optical microscopy and scanning electron microscope( SEM). The results showed that the pileus color of " Huozhi No. 1" was yellowish brown and thicker,while that of wild G. lucidum was mainly reddish brown,the context was thinner,and there were gravel and rotten wood at the bottom of the stipe. A clear skeletal hyphae and binding hyphae were observed in cultivated and wild G. lucidum,but there was no significant difference. The shell layer,context layer,mediostratum layer and spores of cultivated and wild G. lucidum were observed by SEM,and the results showed that there was no significant difference. It was found that the mediostratum of " Huozhi No. 1" was thin and irregular,while the mediostratum of wild G. lucidum was neat and compact. There were two types of spores in wild G. lucidum,one of which retained the outer wall of spore type Ⅰ,with tiny pores on the surface. The other is type Ⅱ spores with many spinous processes on the surface,which may be formed by type Ⅰ spores falling off the outwall. In this study,the appearance characters and internal structure of cultivated and wild G. lucidum in Huoshan were systematically observed and compared,which provided theoretical basis and reference for the identification and quality evaluation of cultivated and wild G. lucidum.

Verification of the theory of "Lieque (LU 7) for the disorders of the head and neck" based on infrared thermography / 中国针灸

OBJECTIVE@#To verify that whether or not through the effects of the externally and internally related meridians in treatment, Lieque (LU 7) is adopted specially for the disorders of the head and neck.@*METHODS@#A total of 36 healthy volunteers were collected from the students of Gansu University of CM and were divided into a Lieque group and a Jingqu group according to the random number table, 18 cases in each one. In the Lieque group, Lieque (LU 7) on the unilateral side was punctured in the subjects. In the Jingqu group, Jingqu (LU 8) was taken as the control because it was located close to Lieque (LU 7) and on the same meridian. Before and after acupuncture in the two groups, separately, the infrared thermography was adopted to determine the temperature changes at the acupoints of the lung meridian of hand-, i.e. Jingqu (LU 8), Lieque (LU 7), Kongzui (LU 6), Chize (LU 5) and Tianfu (LU 3) as well as the acupoints of the large intestine meridian of hand-, i.e., Wenliu (LI 7), Shousanli (LI 10), Quchi (LI 11), Shouwuli (LI 13) and Binao (LI 14).@*RESULTS@#After acupuncture stimulation at Lieque (LU 7), the temperature at the acupoints of the lung meridian of hand-, i.e. Jingqu (LU 8), Lieque (LU 7), Kongzui (LU 6), Chize (LU 5) and Tianfu (LU 3) and the acupoints of the large intestine meridian of hand-, i.e. Wenliu (LI 7), Shousanli (LI 10), Quchi (LI 11), Shouwuli (LI 13) and Binao (LI 14) was all higher obviously as compared with the temperature before acupuncture stimulation (all 0.05).@*CONCLUSION@#For the disorders of the head and neck, acupuncture at Lieque (LU 7) achieves the stimulation and communication of both the lung meridian and the large intestine meridians, so that it is applicable for the disorders of the externally and internally related meridians.

Progresses in therapeutic strategies for thymic rejuvenation / 生理学报

The thymus is a vital primary lymphoid organ that provides unique microenvironments for the proliferation, differentiation, and maturation of T cells. With advancing age, however, the thymus gradually undergoes age-related involution and reduction in immune function, which are characterized by decreases in tissue size, cellularity, and naïve T cell output. This dynamic process leads to the reduced efficacy of the immune system with age and contributes to the increased susceptibility to infection, autoimmune disease, and cancer. In addition, bone marrow transplantation, radio-chemotherapy and virus infection also impair the thymus and give rise to the decline in immune function. Therefore, understanding the molecular mechanisms involved in age-related thymic involution and development of novel therapeutic strategies for thymic rejuvenation have gained considerable interests in recent years. This review emphasizes thymic microenvironments and thymocyte-stromal cell interactions and summarizes our current knowledge about thymic rejuvenation in terms of sex steroid, cytokines, growth factors, hormones, transcription factors, cell graft, and microRNAs. At the end of each discussion, we also highlight unanswered issues and describe possible future research directions.

Microarray-based apoptosis gene screening technique in trichostatin A-induced drug-resisted lung cancer A549/CDDP cells / 解放军医学杂志

Objective: To detect the expression profile changes of apoptosis-related genes in trichostatin A (TSA)-induced drug-resisted lung cancer cells A549/CDDP by microarray, in order to screen the target genes in TSA treating cisplatin-resisted lung cancer. Methods: A549/CDDP cells were treated by TSA for 24 hours. Total RNA was extracted and reversely transcribed into cDNA. Gene expression levels were detected by the NimbleGen whole genome microarray. Differences of expression profiles between TSA-treated and control group were measured by NimbleScan 2.5 software and GO analysis. Apoptosis and proliferation related genes were screened from the expression changed genes. Results: Compared with the control group, 85 apoptosis-related genes were up-regulated and 43 growth or proliferation related genes were down-regulated in the TSA-treated group. GO analysis showed that the functions of these genes are mainly regulating apoptosis, cell resistance to chemical stimuli protein, as well as regulating cell growth, proliferation and the biological process of maintaining the cell biological quality. TSA-activated not only the mitochondrial apoptotic pathways, but also the death receptor related apoptosis pathway, and down-regulated the drug resistance related genes BAG3 and ABCC2. Conclusion: TSA may cause the expression changes of apoptotic and proliferation genes in A549/CDDP cells, these genes may play a role in TSA treating cisplatin-resisted lung cancer.

Cloning and prokaryotic expression analysis of DNA methyltransferas 1(MET1) in Ganoderma lucidum / 中国中药杂志

DNA methyltransferase is the key enzyme in the process of DNA methylation, playing an important role in regulation of gene expression in vivo. According to the Ganoderma lucidum transcriptome data, a full-length cDNA sequence of MET1 from G. lucidum was cloned for the first time, the GenBank registration number is KU239998, and we conducted a comprehensive bioinformatics analysis of the genetic characteristics and spatial structure. The prokaryotic expression analysis showed that E.coli[pET28a(+)-GlMET1] in BL21(DE3) could induce objective protein, shaking the culture at 16 ℃ until the host bacterium(A₆₀₀) was approximately 0.8, and added IPTG to finally concentration of 0.2 mmol•L⁻¹, and then the optimal expression of GlMET1 recombinant protein was accumulated for the induction time of 20 h. The real-time PCR results showed that the expression levels of GlMET1 had obvious differences among varieties of G. lucidum. During the maturity stage, the expression levels of GlMET1 were lower than that in juvenile stage, the results showed that with the growth of G. lucidum, the expression levels of GlMET1 were on the decline. The research provided an important basis for studying the mechanism of DNA methyltransferase thoroughly.

Caveolin-1 and glucose transporter 4 involved in the regulation of glucose-deprivation stress in PC12 cells / 生理学报

Recent evidence suggests that caveolin-1 (Cav-1), the major protein constituent of caveolae, plays a prominent role in neuronal nutritional availability with cellular fate regulation besides in several cellular processes such as cholesterol homeostasis, regulation of signal transduction, integrin signaling and cell growth. Here, we aimed to investigate the function of Cav-1 and glucose transporter 4 (GLUT4) upon glucose deprivation (GD) in PC12 cells. The results demonstrated firstly that both Cav-1 and GLUT4 were up-regulated by glucose withdrawal in PC12 cells by using Western blot and laser confocal technology. Also, we found that the cell death rate, mitochondrial membrane potential (MMP) and intracellular free Ca(2+) concentration ([Ca(2+)]i) were also respectively changed followed the GD stress tested by CCK8 and flow cytometry. After knocking down of Cav-1 in the cells by siRNA, the level of [Ca(2+)]i was increased, and MMP was reduced further in GD-treated PC12 cells. Knockdown of Cav-1 or methylated-β-Cyclodextrin (M-β-CD) treatment inhibited the expression of GLUT4 protein upon GD. Additionally, we found that GLUT4 could translocate from cytoplasm to cell membrane upon GD. These findings might suggest a neuroprotective role for Cav-1, through coordination of GLUT4 in GD.

Based on evidence to establish reporting guidelines for TCM-featured acupuncture systematic reviews/Meta-analysis: a methodology discussion / 中国针灸

Existing reporting guidelines for systematic reviews/Meta-analysis (SRs/MAs) cannot meet the requirements of clinical practice and scientific research, so based on evidence, methods and thoughts on establishing reporting guidelines for TCM-featured acupuncture systematic reviews/Meta-analysis were proposed. Through literature analysis, according to evidence-based principle, preliminary ideas on methodology of establishing reporting guidelines for TCM-featured acupuncture systematic reviews /Meta-analysis were proposed. With consensus as one main research method, it was proposed that in the consensus that was selected and established by experts, the pro- portion of Chinese scholars should be increased to fully declare the opinions of Chinese acupuncturists, and by verification of practice and application, the reporting guidelines for TCM-featured acupuncture systematic reviews/Meta-analysis can be finally made.

Cloning and bioinformatics analysis of chorismate mutase gene from Salvia miltiorrhiza / 中国中药杂志

Chorismate mutase catalyzes the conversion of chorismate to prephenate that is the first committed step in the biosynthesis of the aromatic amino acids phenylalanine and tyrosine. A chorismate mutase gene, designated SmCM1, was isolated from Salvia miltiorrhiza by using RT-PCR. The full length of SmCM1 cDNA consists of 948 nucleotides and has an open reading frame of 765 bp. The deduced amino acid sequence of SmCM1 has 255 amino acid residues which forms a 36.0 kD polypeptide with calculated pI of 6.41 as expected. The putative polypeptide contains a CM_2 super family function domain. Blast W results showed that SmCM1 had 70% of the similarity with Petunia x hybrid CM, 72% of the similarity with Arabidopsis thaliana CM, and 64% of similarity with Populus trichocarpa CM. The transcription level of SmCM1 in root, stem and leaf was analysed by realtime quantitative PCR. The results showed the expression level of the SmCM1 in leaf was highest, and lowest in root. Yeast extract and silver ion joint induction could markedly stimulate the increase of mRNA expression of SmCM1 and its upstream 3-deoxy-7- phosphoheptulonate synthase (DAHPS) and chorismate synthase (CS). It was 7.9, 5.5 and 9.8 times of control on 8 h after induction, respectively.

Cloning and bioinformatics analysis of SmNAC1 from Salvia miltiorrhiza hairy root / 中国中药杂志

In order to study function of NAC transcription in development, hormone regulation and the stress response of Salvia miltiorrhiza, the NAC transcription was cloned and analyzed. By retrieving cDNA database of S. miltiorrhiza hairy root one NAC unigene was found, then a full length of cDNA was cloned by designing specific primers and PCR amplifying. Using ORF finder it was found that the cDNA containing a NAC-AB conserved domain in N-terminal, so the cDNA was a NAC transcription factor, named as SmNAC1 (kF006346). Bioinformatics analysis showed that SmNAC1 had an open reading frame (ORF) of 591 bp encoding 196 amino acids. The calculated protein had isoelectric point (pI) of 4.36 with molecular weight about 21.66 kDa. The transcription level of SmNAC1 after dealing with yeast extract (YE) and silver ion (Ag+) in S. miltiorrhiza hairy root was markedly stimulated up regulating. It was 1.4 fold compared with the control after induction 2 h, and maintained 2.0 fold on 4-12 h after induction. SmNAC1 may participate in regulation of stress response of YE + Ag+.

Pulmonary surfactant protein gene mutation associated with pediatric interstitial lung disease: a case study and the review of related literature / 中华儿科杂志

<p><b>OBJECTIVE</b>To report a case of pulmonary surfactant protein (SP) gene mutation associated with pediatric interstitial lung disease, and study the clinical diagnosis process and review of related literature, to understand the relationship between interstitial lung disease and SP gene mutation in infants and children.</p><p><b>METHOD</b>The clinical, radiological, histological, and genetic testing information of a case of SP gene mutation related pediatric interstitial lung disease were analyzed and related literature was reviewed.</p><p><b>RESULT</b>A 2-year-old girl without a history of serious illness was hospitalized because of the shortness of breath, cough, excessive sputum, and the progressive dyspnea. Physical examination on admission revealed tachypnea, slight cyanosis, and the retraction signs were positive, respiratory rate of 60 times/minute, fine crackles could be heard through the lower lobe of both lungs; heart rate was 132 beats/minute. No other abnormalities were noted, no clubbing was found. Laboratory test results: pathologic examination was negative, multiple blood gas analysis suggested hypoxemia. Chest CT showed ground-glass like opacity, diffused patchy infiltration. Bronchoalveolar lavage fluid had a large number of neutrophils, and a few tissue cells. Eosinophil staining: negative. Fluconazole and methylprednisolone were given after admission, pulmonary symptoms and signs did not improve, reexamination showed no change in chest CT. Then lung biopsy was carried out through thoracoscopy. Histopathology suggested chronic interstitial pneumonia with fibrosis. The heterozygous mutation of R219W in the SFPTA1 and the S186N in SFTPC were identified by SP-related gene sequencing. The review of related literature showed that polymorphisms at the 219th amino acid in SP-A1 allele were found in adults with idiopathic pulmonary fibrosis (IPF), but there is no related literature in pediatric cases. The patient in this report had a mutation at the SP-A1 allele consistent with related literature. Data of 17 young children with mutation in SP-C gene showed that all the 17 cases had dyspnea and tachypnea, chest CT revealed diffuse opacities in lungs, the pathology of lungs was NSIP and CPI. There were 17 kinds of mutation and the common mutation was I73T. The mutation of S186N in SFTPC in our case has never been shown in previously published literature.</p><p><b>CONCLUSION</b>A case of interstitial lung disease with S186N gene mutation in SFTPC was preliminarily diagnosed in an infant. The SP-C gene mutations and polymorphisms are associated with pediatric interstitial lung disease.</p>

Hemorrhoid sclerotherapy with the complication of abdominal compartment syndrome: report of a case / 中华医学杂志(英文版)

The complications of injection sclerotherapy for hemorrhoid are always local. Herein, we report a case in which a female patient with abdominal compartment syndrome developed after receiving a local injection of a sclerosing agent for hemorrhoid.

Roles of Newcastle disease virus in human acute monocytic leukemia in vitro and in vivo / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Some research has shown that Newcastle disease virus (NDV) is effective in the treatment of various tumors, including transferred melanoma and well differentiated renal cell carcinoma. This study aimed to evaluate the effect of NDV against human acute monocytic leukemia SHI-1 cells in vitro and in vivo.</p><p><b>METHODS</b>In vitro, the density and morphologic changes between wild SHI-1 cells (control) and NDV-infected SHI-1 cells were observed. MTT assay was utilized to observe the effect of NDV on the proliferation of SHI-1 cells. In vivo, the effect of NDV on the tumor inhibition was assessed using SHI-1 xenografts subcutaneously established in CD-1 nude mice. NDV was given by intra-tumor injections, and the tumor inhibition rate and toxic effects were evaluated.</p><p><b>RESULTS</b>In the control group, the SHI-1 cells were observed using an inverted microscope to be regular in morphology and intensive in distribution. In the NDV-infected group, the SHI-1 cells were irregular and sparsate, and the aggregate and fused cells were common. MTT assay showed that the proliferation of SHI-1 cells were significantly inhibited by NDV at different concentrations (P<0.01) and in a time- and concentration-dependent manner. The tumor inhibition rate in the NDV group was 84.7%, which was significantly higher than that in the control group (P<0.01). No toxic effects were observed in the nude mice.</p><p><b>CONCLUSIONS</b>NDV can suppress the proliferation of human acute monocytic leukemic cells both in vitro and in vivo. The safety of NDV is reliable.</p>

Strontium fructose 1,6-diphosphate (FDP-Sr) alleviates oligozoospermia induced by multiglycosides of tripterygium wilfordii in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of strontium fructose 1,6-diphosphate (FDP-Sr) on the multiglycosides of tripterygium wilfordii Hook. f. (GTW)-induced oligozoospermia in male rats.</p><p><b>METHODS</b>Forty SD male rats were randomly divided into 3 groups: model (n=10), FDP-Sr (n=10), and control (n=10). The model rats received intragastric administration of GTW at 30 mg/(kg x d) for 40 days to induce oligozoospermia, the rats of the FDP-Sr group orally administered FDP-Sr at 200 mg/(kg x d) for 30 days after GTW induction, while the control rats given but distilled water. Then we obtained the gonad indexes involving the testis, epididymis, preputial gland and seminal vesicle, and determined the count and motility of epididymal sperm, testicular pathomorphology, serum testosterone level and activities of succinodehydrogenase (SDH), lactate dehydrogenase (LDH) and acid phosphatase (ACP) in the testis.</p><p><b>RESULTS</b>The indexes of the testis and seminal vesicle in the control, model and FDP-Sr groups were (0.71 +/- 0.04) and (0.29 +/- 0.04)%, (0.37 +/- 0.04) and (0.25 +/- 0.05)%, and (0.45 +/- 0.07) and (0.31 +/- 0.06)%, respectively; the epididymal sperm counts were (59.87 +/- 11.28), (11.06 +/- 2.53) and (20.95 +/- 4.98) x 10(6)/ml; the serum testosterone levels were (85.31 +/- 7.41), (65.33 +/- 2.90) and (75.32 +/- 5.34) ng/L; and the activities of ACP, LDH and SDH were (95.64 +/- 19.27), (9574.73 +/- 3 578.06) and (6.39 +/- 1.93) U/g prot, (58.42 +/- 12.38), (4820.77 +/- 1 535.22) and (3.48 +/- 0.91) U/gprot, and (83.74 +/- 21.30), (7649.01 +/- 3 123.02) and (5.59 +/- 1.75) U/g prot. All the parameters above were significantly increased in the FDP-Sr group as compared with the GTW models (P < 0.05). Besides, FDP-Sr treatment significantly alleviated the injury of the testicular seminiferous epithelium.</p><p><b>CONCLUSION</b>FDP-Sr can alleviate GTW-induced oligozoospermia, which is closely related with its improvement of testicular function.</p>

Gaussia luciferase reporter assay for assessment of gene delivery systems in vivo / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To develop an alternative method for assessment of gene delivery systems in vivo.</p><p><b>METHODS</b>Mouse primary spleen lymphocytes were genetically modified in vitro by a retroviral vector harboring a Gaussia luciferase (Gluc) expression cassette. After implantation of these cells into recipient mice, the expression of Gluc was detected in whole blood or plasma collected.</p><p><b>RESULTS</b>As little as 10 muL whole blood drawn from the recipient mice could guarantee prompt reading of Gluc activity with a luminometer. And the reading was found in good correlation with the number of genetically modified spleen lymphocytes implanted to the mice.</p><p><b>CONCLUSIONS</b>Gluc may be useful as an in vivo reporter for gene therapy researches, and Gluc blood assay could provide an alternative method for assessment of gene delivery systems in vivo.</p>

Explanatory clinical trials and pragmatic clinical trials on clinical effective evaluation of acupuncture / 中国针灸

The concepts and relations of explanatory clinical trials and pragmatic clinical trials on randomized controlled trials of acupuncture are stated. The explanatory clinical trials and pragmatic clinical trials are different in the purpose of study, the controlled method, the treatment method and the solution problems. The explanatory clinical trials may enhance the understanding of the mechanism of a treatment; the findings of pragmatic clinical trials may be applied immediately in clinical practice. This paper explains that these clinical trials are of great importance to the methodology and practices of clinical effective evaluation of acupuncture based on TCM theory.

Comparison between primary anastomosis after intraoperative colonic defecation and Hartmann procedure in patients with obstructive left colon cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To compare primary anastomosis after intraoperative colonic defecation and Hartmann procedure for obstructive left colon cancer.</p><p><b>METHODS</b>Clinical data of 68 patients who underwent emergent laparotomy for left colon cancer with acute bowel obstruction between January 2000 and January 2008 were analyzed retrospectively.</p><p><b>RESULTS</b>Primary resection and anastomosis with intraoperative defecation was performed in 43 patients and Hartmann's procedure in 25 cases. Patients in both groups were comparable in terms of age, gender, nutritional status, underlying diseases, tumor location and stage, etc. The morbidity and mortality in the two groups were 25.6% vs 28.0% (P=0.761) and 2.3% vs 4.0% (P=0.369), respectively, and the differences were not statistically significant. The length of hospital stay (including first resection operation and second admission for colostomy closure) was (16.6+/-7.8) d in the primary anastomosis group and (24.6+/-9.4) d in the Hartmann procedure group, and the difference was statistically significant (P=0.002). The costs of hospitalization in the two groups were CNY 50,192.8+/-39,727.4 and CNY 58,382.1+/-30,304.9 (P=0.020).</p><p><b>CONCLUSION</b>Primary resection with intraoperative colonic defecation is safe and effective, and should be considered as an alternative to Hartmann procedure for obstructive left colon cancer in selected patients.</p>

Radiofrequency ablation combined with transarterial chemoembolization for unresectable primary liver cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Radiofrequency ablation (RFA) followed by transarterial chemoembolization (TACE) for unresectable primary liver cancer (PLC) has not been widely discussed. In this study, the outcome of the combination of RFA with TACE was retrospectively evaluated.</p><p><b>METHODS</b>From May 2003 to March 2008, 127 consecutive PLC patients with a median age of 56.4 +/- 8.8 years underwent RFA plus TACE. All patients were deemed to have unresectable PLC based on their tumor characteristics. The maximal diameter of the tumor was between 1.5 cm and 10.0 cm. Twenty-six cases with small (<or= 3.0 cm), 33 with medium (3.1 - 5.0 cm), and 68 with large (> 5.0 cm) tumors were included in this study. RFA was performed using a RITA Medical Systems expandable electrode device, which was followed by first-time TACE administration one to two months later.</p><p><b>RESULTS</b>Technical success of RFA was achieved in all 127 patients with no severe treatment-related complications. RFA was performed percutaneously in 16 (13.5%) cases, by laparoscopic approach in 19 (15.7%), and through laparotomy in the remaining 92 (72.4%). RFA response was classified as complete ablation in 48 cases, nearly complete ablation in 28, and partial ablation in 51. The total 1-, 2-, and 3-year survival rates after RFA were 83.1%, 55.7%, and 43.7%, respectively. The survival rates at 3 years were 78.6%, 28.1%, and 0 for complete ablation, nearly complete ablation, or partial ablation groups, respectively. Three-year disease-free survival rates for the complete ablation and nearly complete ablation groups were 50.3% and 21.3%, respectively. RFA response and liver function were significant variables influencing survival time as analyzed using the Cox regression model.</p><p><b>CONCLUSION</b>RFA could be the first-line exterminate treatment for unresectable PLC, and TACE following RFA may assist in eradicating the peripheral viable tissue and micro-metastasis.</p>

Study on key problems in the positive control design methods of acupuncture and moxibustion / 中国针灸

To probe into the design method of the clinical studies with positive control of acupuncture and moxibustion, the several key problems are analyzed, which include the definition of positive control, form of the control, statistical and design types, proper application of positive drugs, and properly use placebo control method to counteract weaknesses of studies, reasonably select observation indexes so as to show characteristics of Chinese medicine. The authors indicate that only fully thinking these key problems, can provide clinical evidence of acupuncture and moxibustion with high quality and convincingness.

Preparation of two antigens--Ag85b and HspX of Mycobacterium tuberculosis H37Rv and the effects of their co-administration with adjuvants in mice / 中国医学科学院学报

<p><b>OBJECTIVE</b>To synthesize two antigens-Ag85b and HspX of Mycobacterium tuberculosis H37Rv with molecular biological methods and to observe their biologic activity after co-administration of adjuvants (aluminum and/or CpG) in mice.</p><p><b>METHODS</b>Recombinant expression plasmids pET30a-Ag85b and pET30a-HspX were constructed. The objective DNA fragments was characterized with restriction enzyme. Then the recombinant plasmids were transformed into E. coli BL-21, and two proteins were expressed by induction of isopropyl beta-D-1-thiogalactopyranoside. After purification with anion exchange column Source30, QHP, and hydrophobic chromatography column, two proteins were identified by amino acid sequencing. After the successful preparation of these two antigens, they were co-administered in mice with adjuvants of aluminum and/or CpG (Ag85b, Ag85b + Al, Ag85b + CpG, Ag85b + Al + CpG; HspX, HspX + Al, HspX + CpG, HspX + Al + CpG); one group received normal saline and served as the control. Splenic lymphocytes were isolated for enzyme-linked immunosorbent spot assay to detect the secreted specific interferon-gamma (IFN-gamma); in addition, lymphocytes proliferation test was performed to observe lymphocytes proliferation after in vitro stimulated with two antigens.</p><p><b>RESULTS</b>The purity of two proteins reached 95% after purification. The N-terminal amino acid sequence (15 aa) of the purified proteins was same as the target sequence. For Ag85b, the secreted specific IFN-gamma from isolated splenic lymphocytes after having been stimulated in vitro with Ag85b (80 microg/ml) remarkably increased in Ag85b + CpG group, Ag85b + Al group, and Ag85b + CpG + Al group; the changes were significantly different between these three groups and control group (P < 0.05). For HspX, the changes were significantly different between HspX + Al + CpG group and normal sodium group, although remarked increase of IFN-gamma was also observed in HspX group, HspX + Al group, and HspX + CpG group.</p><p><b>CONCLUSIONS</b>Ag85b and HspX were successfully expressed and purified. A cell-mediated immunity may be induced when the antigens are co-administered with adjuvants of aluminum and/or CpG in mice, indicating that the recombinant proteins are bioactive.</p>

Characterization of chemical components of essential oil from cupitulum of Chrysanthemum morifolium cultivated in Tongxiang city / 中国中药杂志

<p><b>OBJECTIVE</b>To analyze the chemical components of the essential oil extracted from five cultivars' Hangjuhua (Zaoxiaoyangju, Chixiaoyangju, Dayangju, Yizhongdabaiju and Xiaotangju), which were breed came from the Chrysanthemum morifolium Ramat, and provide scientific basis for quality control.</p><p><b>METHOD</b>The essential oil of five cultivars' Hangjuhua were extracted by water-steam distillation and separated by GC capillary column chromatography. The components were quantitatively determined with normalization method, and identified by GC-MS.</p><p><b>RESULT</b>From the Zaoxiaoyangju, Chixiaoyangju, Dayangju, Yizhongdabaiju and Xiaotangju, 179, 167, 209, 211 and 270 chromatographic peaks were detected, among them 54, 56, 60, 55 and 65 components were identified, which were composed of 73.40%, 64.14%, 65.19%, 53.55% and 62.89% of the total essential oil, respectively.</p><p><b>CONCLUSION</b>The common chemical constituents were juniper camphor, 1, 3, 3-trimethylcyclohex-1-ene-4-carboxaldehyde, borneol, alpha-curcumene, alpha-bisabolol, cis-caryophyllene, benzyl benzoate, methyl beta, beta-dimethylbenzenepropanoic acid ester, 2, 4-decadienal and heneicosane in the essential oil of the five cultivars' Hangjuhua. Among them, juniper camphor is most which were composed of 11.96%, 10.51%, 10.95%, 13.28% and 10.77% of the total essential oil, and methyl beta, beta-dimethylbenzenepropanoic acid ester is second, which were composed of 1.51%, 1.86%, 2.52%, 4.89% and 2.51% of the total essential oil, respectively. There are marked differences in the compounds contained in the five samples. The juniper camphor is a characteristic constituent in the essential oil of Hangjuhua. The resultant data provided the evidences to establish Hangjuhua' cultivars and scientific basis for quality control of Hangjuhua.</p>