1.Lymph node metastatic models of VX2 tumor in New Zealand white rabbits and their biological characteristics
Yiqun SUN ; Tong TONG ; Jian MAO ; Fangfang ZHONG ; Yajia GU
China Oncology 2016;26(10):840-847
Background and purpose:The lymph node metastatic model of rectal tumor is a useful tool for the research on tumor occurrence, development, metastasis and antineoplastic therapy. There are few reports about establishment of larger animal model. This study aimed to establish feasible and reproducible lymph node metastatic models of VX2 tumor in rabbits.Methods:The VX2 tumor tissue was put into the puncture needle. The VX2 tumor tissue in the needle was orthotopically transplanted into the rectal wall of the New Zealand white rabbits successfully. Twenty New Zealand white rabbits were transplanted. Two experimental rabbits were scanned by MR weekly. Tumor growth curve and lymph node numbers were observed on MR. Experimental rabbit tumor volumes were measured by MR post-processing software. The rectal tumor and surrounding lymph nodes were resected, and the specimens were ifxed. The sections were stained with HE. We explored the relationship between tumor volume and growth time, the number of metastatic lymph nodes and tumor volume, respectively.Results:Thirteen models were successfully established with a rate of 65%. Tumors limited in the rectal wall were observed on the fourth week. Tumor size increased over time. There was significant difference in the tumor volume between different periods (growth cycle number) (F=52.865,P<0.05). There was a signiifcantly positive correlation between tumor volume and the growth cycle number (r=0.910,P<0.05). The metastatic lymph nodes could be observed when VT>9 cm3. The number of metastatic lymph node increased obviously from the ninth week. The more tumor volume, the greater the number of metastatic lymph nodes was observed (F=92.531,P<0.05). There was a signiifcantly positive correlation between the number of metastatic lymph nodes and the tumor volume (r=0.945,P<0.05).Conclusion:Metastatic lymph node models of VX2 tumor in New Zealand white rabbits were established successfully. This model has some value in the research on local growth, invasion mechanism, lymph node metastasis and biological characteristics of rectal cancer.
2.Medium Optimization for Antitumor Agent Mycoepoxydiene by Marine Lignicolous Fungi Diaporthe sp.
Ruo-Yu WANG ; Yao-Jian HUANG ; Zhong-Hui ZHENG ; Wen-Jin SU ; Yue-Mao SHEN ;
Microbiology 1992;0(05):-
Mycoepoxydiene is a novel antitumor agent extracted from marine lignicolous fungi HLY-2, which is Diaporthe phaseolorum by molecule identification. The medium optimization for mycoepoxydiene by orthogonal design and the comparison of submerged fermentation and solid state fermentation were studied. The rusult is that the maximal yield of the compound is 543mg/L, which is 43 times compared to the customary half-seawater PD medium and 15 times to the best submerged condition. This optimum culture medium included potato 250g/L, seawater 300mL/L, glucose 30g/L, lactose 50g/L, KH_ 2 PO_ 4 0.65mmol/L and (NH_ 4 )_ 2 SO_ 4 1g/L in the solid state condition. Differentiation analysis between submerged and solid state fermentation, and antitumor activity of these ferment products were also studied. The antitumor activity of products of the optimum medium approached the pure compound.
3.Genetic Engineering Reconstruction of Klebsiella pneumoniae Producing 1,3-propanediol by the Gene yqhD Encoding 1,3-propanediol Oxidoreductase Isoenzyme
Bin ZHUGE ; Yong WANG ; Hui-Ying FANG ; Zhong-Gui MAO ; Jian ZHUGE ;
China Biotechnology 2006;0(11):-
In the reductive branch, glycerol is first dehydrated to 3-hydroxypropionaldehyde that is then reduced to 1,3-PD under the consumption of reducing power (NADH). If over-expression of the gene dhaB encoding glycerol dehydrase is achieved,the reducing power will be scarce and 3-hydroxypropionaldehyde will be accumulated,which is disadvantage to produce 1,3-propanediol.The structure gene yqhD from E.coli encoding 1,3-propanediol oxidoreductase isoenzyme[under the consumption of reducing power (NADPH)]and the gene dhaB encoding glycerol dehydrase from Klebsiella pneumoniae was amplified using PCR method.The two gene were transferred into expression vector pEtac to construct a novel recombinant Klebsiella pneumoniae (pEtac-dhaB-tac-yqhD).Over-expression of yqhD and dhaB in Klebsiella pneumoniae was achieved with pEtac-dhaB-tac-yqhD.The fermentation result on aerobic conversion showed the increase of 20% of 1,3-propanediol yield by Klebsiella pneumoniae(pEtac-dhaB-tac-yqhD) was obtained compared with Klebsiella pneumoniae.The main by-products,acetic acid and butanediol decreased estrogen receptors 30% obviously.
4.Development of a Multiplex PCR-Microarray Method for Detection of Important Enteropathogen
Yuan-Hai YOU ; Xun ZENG ; Wei GUO ; Yan YIN ; Mao-Jun ZHANG ; Jian-Zhong ZHANG ;
China Biotechnology 2006;0(12):-
Objective: To establish a multiplex PCR-microarray method for detecting important enteropahogens.Methods: Uniplex and multiplex PCR were performed to obtain the best primer sets for identifying the target bacteria at species and multi-species level.Fluorescent dyes were mixed into PCR reaction to determine whether it can affect the efficiency of amplification.To improve the efficiency of microarray,a 35 pairs primer-labeling system was optimized based on the hybridization results to find the best combination to avoid false negative results.Results: Specific PCR products were all obtained using species-specific primer sets.More preferential amplification may happen when more primer pairs were added to the reaction.The hybridization results showed a positive association between the efficiency of multiplex-PCR and signal intensity.Conventional PCR yielded more products than fluorescent dyes labeled PCR.Thirty-five primers were divided into three different combinations to label target respectively,hybridization results showed a high specificity.Conclusion: Mixing fluorescent dyes into PCR may reduce the efficiency of amplification and hybridization,but may have no effect on the analysis of hybridization results.The hybridization efficiency of microarray depends on the amplification efficiency of multiplex PCR.For microarray target labeling,three primer sets could be used to avoid negative hybridization led by preferential amplification of multiplex-PCR.It indicates that the multiplex PCR-microarray method is an attractive diagnosis tool for the high-throughput identification of enteropathogenic organisms especially for multiple causative agents and epidemiological investigations.
5.Puerarin inhibits DNA damage of HaCaT cells induced by UVB via ceramide pathway.
Mian-Qing HUANG ; Zhao-Xin YANG ; Mao-Zhong YAO ; Shu-Hong TIAN ; Jian FU
Acta Pharmaceutica Sinica 2012;47(5):609-613
This study is to investigate the effect and mechanism of puerarin on DNA damage of HaCaT cells induced by UVB. Puerarin pre-treated cells were irradiated with UVB at 30 mJ x cm(-2). Twenty four hours after irradiation, DNA damage was detected by comet assay, ceramide was measured by thin layer chromatography and gas chromatography, intracellular free calcium ion was analyzed by flow cytometry, the phosphorylation level of p38 protein was examined by Western blotting method. Levels of DNA damage, ceramide, free calcium ion and p-p38 protein were elevated in UVB model cells. Contrary to the model group, all indicators above were reduced in all groups pre-treated by puerarin. Puerarin restrains the ceramide accumulation to block downstream p38 MAPK pathway and calcium ion rising, therefore reduces DNA damage in HaCaT cells induced by UVB.
Calcium
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metabolism
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Cell Line
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Ceramides
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metabolism
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DNA Damage
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drug effects
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radiation effects
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Down-Regulation
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Humans
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Isoflavones
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pharmacology
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Keratinocytes
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cytology
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metabolism
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Phosphorylation
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Signal Transduction
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drug effects
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Ultraviolet Rays
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adverse effects
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p38 Mitogen-Activated Protein Kinases
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metabolism
7.Microbial degradation of aniline by bacterial consortium.
Jian-Long WANG ; Ze-Yu MAO ; Wei-Zhong WU
Biomedical and Environmental Sciences 2003;16(4):398-404
OBJECTIVETo investigate the characteristics of microbial degradation of aniline by a stable bacterial consortium.
METHODSThe bacterial consortium was isolated from activated sludge treating chemical wastewater using aniline as the sole source of carbon and nitrogen by enrichment and isolation technique. The biomass was measured as optical density (OD) at 510 nm using a spectrophotometer. Aniline concentrations were determined by spectrophotometer. The intermediates of aniline degradation were identified by GC/MS method.
RESULTSThe bacterial consortium could grow at a range of aniline concentrations between 50 and 500 mg/L. The optimal pH and temperature for aniline degradation were determined to be 7.0 and 30, respectively. The presence of NH4NO3 as an additional nitrogen source (100-500 mg/L) had no adverse effect on bacterial growth and aniline degradation. The presence of heavy metal ions, such as Co2+, Zn2+, Ni2+, Mn2+ and Cu2+ had an inhibitory effect on aniline degradation.
CONCLUSIONSThe isolated bacterial consortium can degrade aniline up to 500 mg/L effectively and tolerate some heavy metal ions that commonly exist in chemical wastewater. It has a potential to be applied in the practical treatment of aniline-containing wastewater.
Aniline Compounds ; metabolism ; Bacteria ; Biomass ; Bioreactors ; Carcinogens ; metabolism ; Chemical Industry ; Hydrogen-Ion Concentration ; Metals, Heavy ; analysis ; Waste Disposal, Fluid ; methods ; Water Pollutants ; metabolism
8.Treatment of infantile parotid hemangioma with propranolol.
Xue-jian LIU ; Zhong-ping QIN ; Mao-zhong TAI ; Ke-lei LI ; Chun-xiao GE
Chinese Journal of Stomatology 2010;45(5):292-294
OBJECTIVETo evaluate the short-term results and safety of propranolol for the treatment of infantile parotid hemangioma.
METHODSOral propranolol was administered to 17 infants with parotid hemangioma at a dose of 1.0-1.5 mg per kilogram of body weight per day. The patients were revisited once a week. The changes of the tumor size, texture and colour were monitored and recorded at a regular interval. The adverse effects after medication were observed and managed accordingly. The short-term results were evaluated using a 4 scales system.
RESULTSAmong the 17 patients treated, the follow-up time was 5 to 10 months. The overall response was scale I in 0 patient, scale II in 0 patients, scale III in 5 patients, and scale IV in 12 patients. No serious adverse effects were encountered.
CONCLUSIONSOral propranolol at a lower dose is a safe and effective method for the treatment of infantile parotid hemangioma. The short-term results were excellent and the side effects minimal.
Administration, Oral ; Adrenergic beta-Antagonists ; administration & dosage ; therapeutic use ; Female ; Follow-Up Studies ; Hemangioma ; drug therapy ; Humans ; Infant ; Male ; Parotid Neoplasms ; drug therapy ; Propranolol ; administration & dosage ; therapeutic use ; Treatment Outcome
9.Treatment of infantile vascular malformations associated with airway obstruction.
Xue-jian LIU ; Zhong-ping QIN ; Mao-zhong TAI ; Ke-lei LI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2010;45(1):32-37
OBJECTIVETo summarize the clinical features of vascular malformations complicated with airway obstruction and to evaluate the therapeutic methods of these disease.
METHODSForty-seven children with airway obstruction and dyspnea (25 males, 22 females) were treated from Jun 1985 to Dec 2007, and their clinical data were retrospectively analyzed. Among 47 patients, there were 27 cases of venous malformations, 17 cases of macrocystic lymphatic malformations, and 3 cases of microcystic lymphatic malformations. Injection with absolute alcohol were performed in 20 patients with venous malformations, whereas both surgery and injection were performed in 7 patients with extensive or multiple lesions. Seventeen patients with macrocystic lymphatic malformations were treated with pingyangmycin injection. While surgery combined with pingyangmycin injection were used in other 3 patients with microcystic lymphatic malformations. According to the degree of airway obstruction and therapeutic conditions, tracheal intubation was performed in 27 patients, urgent preoperative tracheotomy was performed in 3 patients, prophylactic tracheotomy was performed in 2 patients, and postoperative tracheotomy was performed in 1 patient.
RESULTSTracheal intubation was remained for 24 to 48 hours in 30 patients, whose intubation was removed successfully in 29 patients except 1 patient who occurred dyspnea after removal of tracheal intubation resulting in tracheotomy. Tracheal cannula was successfully removed in all 6 patients 3 weeks to 4 months after the tracheotomy. There were 9 patients treated once, whereas injections were repeated 2 to 5 times in 38 patients. Necrosis of mucosa occurred in 2 cases after the injection with absolute alcohol, while temporary hemoglobinuria one occurred in 1. There were 5 cases of light or mediate fever after the pingyangmycin injection who recovered well after the symptomatic treatment. Follow-up lasted 1 to 23 years, 38 patients cured, 9 patients valid, and no patient invalid.
CONCLUSIONSIt is suggested that sclerotherapy should be the first choice in the treatment of vascular malformations complicated with airway obstruction, in which absolute alcohol should be used in venous malformations compared to pingyangmycin in lymphatic malformations. Combined therapy should be carried out in patients with extensive lesions in order to shorten the course of treatment and to get good therapeutic result.
Adolescent ; Airway Obstruction ; complications ; Child ; Child, Preschool ; Combined Modality Therapy ; Female ; Humans ; Infant ; Male ; Treatment Outcome ; Vascular Malformations ; complications ; therapy
10.A survey of epidemiological factors of coal-burning-borne endemic fluorosis in northeast of Chongqing
Ju, YAN ; Zhao-hui, ZHONG ; Ying-xiong, WANG ; Li, WANG ; Kai-wen, YONG ; Mao-zhong, ZHANG ; Xing-jian, LUO ; Wei, YAN
Chinese Journal of Endemiology 2012;31(6):653-656
Objective To explore the risk factors of coal-burning-borne endemic fluorosis (endemic fluorosis) and to provide a scientific basis for making prevention and control strategy.Methods One hundred children with dental fluorosis and 30 adults with skeletal fluorosis who lived in Wushan county and Fengjie county,the endemic fluorosis regions northeast of Chongqing were randomly selected as the case group in 2010.One hundred healthy children and 30 healthy adults were selected from the population where the cases were selected from as the control group and 30 healthy external adults control were selected from Yubei district,a non-endemic fluorosis area.Fasting venous blood was draw from all subjects and the content of zinc (Zn),copper(Cu),calcium (Ca),magnesium(Mg),and ferrum(Fe) in venous blood was measured by atomic spectrophotometric.Instant urine was collected from all subjects and urinary fluorine was measured by fluorine selective electrode.We also did questionnaire survey to 100 dental fluorosis children and 100 control children about their situation of fluorine pollution and knowledge related to endemic fluorosis.Results Children's blood Zn of the ease group[(70.88 ±9.28) μmol/L] was lower than that of the control group [(75.53 ± 10.78)μmol/L],and the difference was statistically significant(P < 0.05).But the average blood Cu content[(30.28 ± 2.58)μ mol/L] and the average urinary fluorine of the case group [(0.74 ± 0.36)mg/L] were significantly higher than that [Cu (28.45 ± 4.05)μmol/L and urinary fluorine (0.48 ± 0.21)mg/L] of the control group in children,respectively,and the differences was statistically significant(P < 0.05).The average blood Zn content of adults in the case group[(91.13 ± 10.29)μ mol/L] was lower than that of the control group[(99.57 ± 11.73)μmol/L],and the difference was statistically significant (P < 0.05).The average content of Mg[(1.57 ± 0.19)mmol/L],Fe[(8.17 ± 1.01) mmol/L] and urinary fluorine[(2.37 ± 1.01)mg/L] in the case group were higher than that [Mg(1.46 ± 0.16) mmol/L,Fe(7.72 ± 0.96) mmol/L and urinary fluorine(0.92 ± 0.85)mg/L] of the control group,respectively,and the differences were statistically significant(P <0.05).In the questionnaire survey,we found that the following were important related factors (OR values were 2.7335,0.3339,2.8428,0.4633,0.5439,0.4009,0.4805 and 0.3994,P < 0.05) between the case group and the control group:using local coal for heating,furnaces and stoves improvement,eating local coal fire baked food,knowing the harm of endemic fluorosis to the health of human body,knowing endemic fluorosis can be prevented,knowing consuming baked food can cause endemic fluorosis,knowing drinking milk can prevent endemic fluorosis and having the habit of eating calcium tablets.Conclusions In addition to the main pathogenic element of fluorine,body Zn content is closely related to the process of coal-burning-borne endemic fluorosis in the two counties,fluorosis patients have lower blood Zn levels,Zn deficiency is an auxiliary factor in the etiology of fluorosis.Reduce the use of local coal for heating,do not consume the baked food,improve stoves,learn more knowledge about endemic fluorosis and ingest anti-fluoride elements can reduce the risk of endemic fluorosis.