This research uses six Agrobacterium rhizogenes R1601, R15384, R1000, A4, R1025 and R1 to infect silymarin explants to induce hairy roots and silibin. All of the six A. rhizogenes can induce Silybum marianum to generate hairy roots and the A. rhizogene A4 shows comparatively high infection on the plant. This research determines the condition to induce silymarin hairy roots by the factors of infection time, pre-culturing, co-culturing and pH value. The fact that MS liquid medium fits the proliferation of silymarin hairy roots is determined. Through PCR molecular identification, it can be seen that the DNA plasmids in the A. rhizogenes are successfully integrated into the genome of transformed roots. Using liquid chromatography, it is determined that the silibin content in silymarin hairy roots is 2.5 times that in the plant In this research, the silymarin hairy roots culturing system is established, which lays a foundation for the study of culturing silymarin hairy roots and producing silibin.
Agrobacterium ; genetics ; physiology ; Cell Culture Techniques ; methods ; Milk Thistle ; chemistry ; genetics ; growth & development ; microbiology ; Plant Roots ; chemistry ; genetics ; growth & development ; microbiology ; Silymarin ; analysis ; Transformation, Genetic

Objective:To observe the effect of rapid point pressure on clinical effect and pulmonary function of patients with chronic persistent bronchial asthma.
Methods:A total of 60 confirmed chronic persistent bronchial asthma cases were randomly allocated into two groups, 30 in each group. Cases in the treatment group were treated with rapid point pressure, 1 h for the initial treatment, and 40 min for the ensuing treatments. The treatment was done once a day for 40 d. Cases in the control group were treated with Compound Methoxyphenamine Hydrochloride Capsules, 2 capsules for each dose, 3 times a day. The treatment lasted for 7 consecutive days. A 1-year follow-up was made for both groups.
Results:After treatment, the scores of each symptom and total symptom scores for the traditional Chinese medicine (TCM) symptoms in the treatment group were significantly higher than those in the control group (P<0.05);the total effective rates in the treatment group were higher than those in the control group in each time frame; and there was a between-group statistical significance in total effective rate after 7 d of treatment (P<0.01). After 40-day treatments, there were statistical significances in the forced expiratory volume in 1 s percentage of predicted value (FEV 1%) and inspiratory capacity (IC%) in the treatment group (P<0.05); however, there were no statistical significances (P>0.05) in forced expiratory volume in 1 second to forced vital capacity ratio (FEV1/FVC), peak expiratory flow rate (PEF%) and maximum midexpiratory flow rate (MMEF%); and there were no statistical significances in pulmonary function parameters in the control group. In addition, as for pulmonary function parameters, there were no intra-group statistical significances in differences before and after treatment (P>0.05).
Conclusion:Rapid point pressure can alleviate patients’ TCM symptoms and improve their FEV%and IC%.

The effect of initial substrate concentration on the growth,metabolic activities of Photosynthetic bacteria (PSB) in the process of hydrogen production is studied.The empirical relation of the initial substrate concentration to the specific growth rate,specific substrate consumption rate and specific hydrogen production rate of PSB are developed based on the modified Monod model.It is found that the results given by the model are well agreed with the experimental data in growth phase and stationary phase of PSB.Meanwhile,the optimal substrate concentration in the process of growth,metabolism and hydrogen production is 50 mmol/L.Furthermore,when the initial substrate concentration deviates 50 mmol/L,the activation of PSB is limited or inhibited,and the inhibiting effect is more prominent than the limiting effect.In addition,it is demonstrated that initial substrate concentration has little effect on the specific substrate consumption rate.

Objective To investigate the expression of caspase-10 in differentiated thyroid carcinoma and association with its development and metastasis. Methods Thyroid samples from 37 patients in a period from January 2006 to December 2007, with differentiated thyroid carcinoma were retrospectively analyzed for caspase-10 by immunohistocbemistry(streptavidin-perosidase, S-P), compared to control group of 46 cases with nodtdar goiter. The relationship between the expression of caspase-10 and the clinical pathologic characteristics of thyroid carcinoma were also explored simultaneously. Results caspase-10 were observed as brown or yellow particles located in the cytoplasm or cell membrane of nodular goiter but there were no significant evidence for its positive expression in thyroid carcinoma, caspase-10 expression was markedly down-regulated in differentiated thyroid carcinoma(29.73%,11/37) compared with benign nodules(71.74%,33/46, χ2=14.528, P<0.01). The positive expression in 18 cases with lymph node metastasis(11.11%,2/18) was significantly lower than those in 19 patients without lymph node metastasis(47.37%,9/19; χ2=4.210, P<0.01). There was no significant correlation(P> 0.05) between the expression of caspase-10 and the clinical pathologic characteristics including male, age, TNM stage and pathologic type. Conclusion Down-regulation of caspase-10 may play a critical role in carcinogenesis and development of differentiated thyroid carcinoma.

OBJECTIVETo observe interventional effects of anti-viral therapy and Compound Qin-gre Granule (CQG) on host cellular immune functions of acute virus infection patients.

METHODSThirty acute virus infection patients were recruited to detect peripheral lymphocyte subsets. They were randomly assigned to two groups, the Western medicine treatment group (treated with anti-virus Western medicine) and the integrative medicine treatment group (treated with anti-virus Western medicine plus CQG). T-cell subsets were re-examined 7 days later. Changes between before and after treatment were observed. Effect on host cellular immune functions and efficacy were compared between the Western medicine treatment and the integrative medicine treatment.

RESULTSCompared with the normal control group, the percentage of peripheral T cells increased, and the percentage of B/NK cells decreased in acute virus infection patients (P < 0.01). Meanwhile, in T cell subsets, the percentage of CD8+ T cells and CD8+ CD38+ T cells increased (P < 0.05, P < 0.01); and percentages of CD4+ T cells, CD4+ CD28 + T cells, and CD8+ CD28+ T cells decreased (P < 0.05, P < 0.01). After one-week treatment, percentages of CD4+ T cells, CD4+ CD28+ T cells, and CD8+ CD28+ T cells increased (P < 0.05, P < 0.01), while the percentage of CD8+ CD38+ T cells decreased (P < 0.01). More significantly, these changes were greater in the integrative medicine treatment group than in the Western medicine treatment group (P < 0.05).

CONCLUSIONSDisarranged cellular immune functions existed in acute virus infection patients. CQG could significantly improve viral infection induced immunologic derangement and immunologic injury.

Objective:To evaluate the accuracy and feasibility of sentinel lymph node biopsy (SLNB) marked by 99Tcm-IT-Ritux-imab and to discuss the clinical value of the method in diagnosis and treatment of cutaneous melanoma. Methods:A total of 67 patients with cutaneous malignant melanoma received 99Tcm-IT-Rituximab-tagged SLNB from March 2008 to March 2012. Lymphoscintigra-phy was conducted 30 min to 1 h after intra-dermal injection of 99Tcm-IT-Rituximab. Subsequently, the surgery of SLNB was carried out using gamma probe. The detection and positive rates of SLNB were counted. The relationship between the status and the clinical features of the sentinel lymph node (SLN) was analyzed, such as the T stage, ulceration, age, gender, and location. The influence of SLN status on overall survival (OS) and disease-free survival (DFS) was evaluated. Results:SLNs were detected in all the 67 patients by SPECT and gamma detector, with detection rate of 100%. Fifteen patients had SLN metastasis, and the positive rate was 22.4%. Chi-square indicates that SLN metastasis is associated with age, T stage, and ulceration (P<0.05). A total of 63 patients were followed up for 24-69 months, and the median follow up time was 43 months. Kaplan-Meier survival analysis shows that both OS and DFS in the SLN-negative group are better than those in the SLN-positive group (OS:93.9%vs. 57.1%, P<0.01;DFS:79.6%versus 28.6%, P<0.01). Cox-regression multiple factors analysis suggests that both SLN status and T stage are independent factors that affect the DFS of malignant melanoma. Conclusion:SLNB assisted by 99Tcm-IT-Rituximab can well reflect the state of lymph node metastasis and is es-sential for accurate staging, prognosis judging, and treatment guiding. Its operation procedure is simple with high accuracy, and the im-aging status is stable. Therefore, it is convenient and feasible as a means of SLNB.

Objective To investigate the cause,prevention methods and curative effect on two different drainage methods of subcutaneous effusion after breast cancer surgery.Methods The clinical data of 68 cases subcutaneous effusion among 2 100 cases underwent breast cancer modified radical surgery in our hospitial were analyzed retrospectively from January 2010 to December 2016.The vacuum sealing drainage was performed in 1 032 cases and wound high negative pressure drainage system was performed in 1068 casess.Both the elastic bandage compresstion dressing didn't used.Results In 2 100 cases,68 cases of subcutaneous effusion occured (3.2％),including longitudinal incision 48 cases and transverse incision 20 cases.Among 1 068 cases of high negative pressure drainagesystem,subcutaneous effusion occured in 36 cases including 28 cases of incisional infection and 8 cases of skin flap necrosis.The extuhation time was 8-36 days,average 12 days.Among 1 032 cases of vacuum sealing drainage,subcutaneous effusion occured in 32 cases including 22 cases of incisional infection,10 cases of skin flap necrosis.The extubation time was 6-12 days,average 8 days.All cases were followed up for 3 months,no subcutaneous effusion and axillary effusion occurred.Conclusions Adequate drainage is the key to prevent subcutaneous effusion.Both vacuum sealing drainage and wound high negative pressure drainage can effectively reduce the occurrence of subcutaneous effusion,however,vacuum sealing drainage is more suitable for wounds with more exudation,larger lacuna and deeper incisions,especially the infected wound and abscess.

Objective:To explore the outcome of soft tissue sarcoma (STS) on patients with soft tissue metastasis. Methods:We ana-lyzed 25 STS patients with soft tissue metastasis primarily localized on extremity and trunk. The study was conducted from June 2010 to June 2016 by retrospective analysis of the clinical and pathological characteristics of the patients. The assessed endpoints were overall survival. Results:Six patients (24%) had synchronous soft tissue metastasis, and 19 patients (76%) had metachronous metasta-sis. The average time for primary tumor recession of metastatic lesions was 45.3 months. Metastases were most common in parts of the trunk in 18 patients (72%), followed by the head and neck in 5 patients (20%). Eleven patients (44%) with lung metastasis had poor prognosis. Conclusion:STS occurred more rarely in soft tissue metastasis than in pulmonary metastasis. Neoadjuvant chemotherapy and surgical treatment were the major therapies employed. Targeted therapy as a new treatment rendered good results.

BACKGROUND: In the past, the incidence rates of infectious diseases,natural epidemic diseases and endemic diseases in three gorges areas were always high. Because the flooded areas of Kaixian was enormous, the population of Kaixian was intensive density, the pollution may harm to the population health after the dam has been established.OBJECTIVE: To predict the influence of the Three-gorges dam on population health in Kaixian County of Chongqing and put forward to scientific evidence of preventive measures through analyzing the main health indicators and influence factors in Kaixian.DESIGN: Cross-sectional study.SETTING: Public Health College of Chongqing Medical University and Disease Controlled Center in Kaixian County of Chongqing.PARTICIPANTS: The survey was conducted in Kai County of Chongqing,from February to May 2004. All disease surveillance data of people in this county was investigated and other people lived in three-gorges area or around area were selected as the controls.METHODS: Collect and sort relevant indicators of social development,population health and public health system of Kaixian and related area to go deep into the representative townships to do some filed investigation,collect water samples of local drainage areas to carry out laboratory quantitative analysis; carry out comparative analysis based on the relevant materials; carry out comprehensive predicting and evaluation to the potential impact on population health of the three gorges engineering by combining the quantitative and qualitative analysis methods.general people in Kai county, Chongqing city and all over the nations durKai county, Chongqing city and all over the nations during 1999 and 2003;ty, Chongqing city and all over the nations during 1999 and 2001: Mortality rates of infants and under-five children were also higher than that average levels of whole country and Chongqing respectively; Multi-indicators showed that the general population health status of Kaixian was great bad,which stood a distinct gap among average levels of the whole country and Chongqing maintained about 250/100 000 or so, which was higher than the national level (190/10 000), however, the incidence rate of infectious diseases of Kaixian was rising year after year, even that of the recent two years has been reached up to more than 1.5 times higher than that of the whole of the water-based infectious diseases, especially the virus hepatitis, was severe. Another potential impact on population health of three-gorge dam is the emergence of natural epidemic diseases and insect-based infectious diseases. Malignant tumor such as hepatocarcinoma.CONCLUSION: We should strengthen current public health system, and cognition to the possible threat on disease epidemic and public health.With the construction of three-gorge dam, it is necessary to speed up establishment of effective measures to reduce predictable influence on population health.

Objective To investigate the biological function of M122 in pathogenesis of MCMV in developmental brain disorders and brain damage, screening for mouse brain cDNA library interacting with M122 was performed by a yeast two-hybrid system. Methods The reconstructed bait plasmid pGBKT7-M122 was transformed into yeast cells AH109 and screened on the nutrient deficiency medium SD/-Trp. After express of the bait protein in AH109 yeast strains was detected by Western blot analysis, yeast-two hybrid screening was performed by mating AH109 with Y187 containing mouse brain cDNA library plasmid. The diploid yeast cells were plated on the nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade. The second screening was performed with SD/-Trp/-Leu/-His/-Ade containing X-α-gal. The plasmids in positive colonies were extracted and transformed into E. coli JM109 cells. After plasmid DNA in JM109 cells were extracted form positive colonies and sequenced, the results were analyzed by bioinformatic methods. The interactions between M122 protein and the protein obtained from positive colonies were further confirmed by repeating yeast-two hybrid. Then, autoactivations of the proteins obtained from positive colonies were detected.Results The reconstructed bait plasmid was transformed into yeast cells AH109 successfully. The bait protein expressed in the yeast cells AH109 stably. 24 proteins interacting with MCMV M122 were screened, including syntaxin 8 ( Stx8 ), phosphoglucomutase 2 ( Pgm2 ), potassium voltage-gated channel, shaker-related subfamily, beta member 1 ( Kcnab1 ), collagen, type ⅪⅩ, alpha 1 ( Col19a1 ), archain 1 ( Arcn1 ), cytidylate kinase( Cmpk), DnaJ(Hsp40) homolog, subfamily A, member 1 (Dnaja1), ATPase, Na+/K + transporting, beta 3 polypeptide( Atp1b3 ), SH3-domain GRB2-like ( endophilin ) interacting protein 1 ( Sgip1 ),ankyrin repeat domain 17 (Ankrd17), Smg-7 homolog, nonsense mediated mRNA decay factor(Smg7),sperm associated antigen 9 ( Spag9 ), FK506 binding protein 1a ( Fkbp1a), MYST histone acetyltransferase monocytic leukemia 4 ( Myst4), hyaluronan and proteoglycan link protein 1 ( Hapln1), autophagy-related 3 (Atg3), splicing factor, arginine/serine-rich 5 ( Sfrs5 ), zinc finger, C3HC-type containing 1 ( Zc3hc1 ),thioredoxin-related transmembrane protein 1 ( Txndc1 ), adaptor protein complex AP-1, gamma 1 subunit (Ap1g1), Cullin 1 ( Cul1 ), and so on. Three of them were formerly unknown proteins. M122 protein could interact with the proteins obtained from positive colonies in the yeast cells AH109. Ap1g1 and Cul1 were proved to have autoactivation. Conclusion A class of proteins in brain interacting with M122 has been obtained. It is presumed that these proteins are correlated with neuropathogenesis of the brain disorders caused by CMV, but the candidates still need further confirmation for the interaction.