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 summarize the experience of long-term survival of the patients under- going orthotopic heart transplantation.Methods Heart transplantation was performed on 2 cases of dilated cardiomyopathy and one case of Keshan disease.Before operation,pulmonary artery pressure was 42-53 mm Hg(5.60-7.07 kPa)and pulmonary vascular resistance 5.6-7.0 wood.The body weight difference between donors and receptors was 10%-15%.There were three same antigens in HLA zygosity experiment for all of three patients.Two cases were subjected to standard heart trans- plantation and one case to whole heart transplantation.All the atriums and big vessels were sutured by evting suture method.Cyclosporin A,azathioprine and corticosteroid were used to prevent patients from rejection.Results Survival time of 3 patients was 13 years and 10 months,12 years and 10 years and 3 months.Heart functions of three patients were NYHAⅠand all of 3 patients are living and working commonly.Six,3 and 1 rejection(s)occurred in 3 patients respectively and cured by appro- priate treatment.Electrocardiogram revealed that case 1 and case 2 had two P waves and case 3 sinus rhythm.Ultrasonic cardiogram showed that in case 1 and case 2,the left and right atriums were enlarged and tricuspid valve had slight backstreaming,and in case 3,all of the cardiac chambers were normal and had no backstreaming of tricuspid valve and mitral valve.No abnormal findings were found in 3 cases by 4-9 times of coronary arteriongraphy.Conclusion The important factors for the patients' long-term survival after heart transplantation include the choice of appropriate donors and acceptors, protection of donors' hearts,selection of appropriate operations and suture methods,rational use of im- munosuppressants and prevention of cardiac allograft vasculopathy.

Objective To analyze the effect of prevention and treatment with lamivudine in HBsAg positive renal allograft recipients with HBV recurrence.Methods In 28 patients with chronic renal failure whose HBsAg was positive,8 cases were positive for HBV-DNA positive.All these 28 cases had normal liver function without hepatic cirrhosis before renal transplantation.All donors were negative for HBsAg.Twenty patients received lamivudine prophylactic treatment:14 cases positive for HBsAg but negative for HBV-DNA before transplantation received lamivudine treatment immedia- tely after transplantation and 6 cases positive for both HBsAg and HBV-DNA received lamidudine treatment before transplantation.Eight patients were treated with lamivudine when their hepatic dys- function with recurrent HBV antigenemia were developed after transplantation.Lamivudine was orally taken and its initial dosage was 100 rag/day.Results The follow-up period of the 28 patients were 13- 54 months with the average of 23.6 months,and 2 died during this period.Mild hepatic dysfunction with recurrent HBV antigenemia developed in 3 of 20 hepatitis antigenemia patients received lamivu- dine prophylactic treatment with a mean duration of 9.3 months after transplantation.The highest average level of serum ALT was 87.5 U/L.The liver function returned to the normal with HBV-DNA negative after lamivudine treatment in 3 patients.The other 17 patients maintained normal liver func- tion with HBV-DNA negative during the follow-up period.Hepatic dysfunction with recurrent HBV antigenemia(or HBV-DNA titer increased significantly)developed with a mean duration of 4.6 months in all 8 patients without receiving lamivudine prophylactic treatment.The highest average level of serum ALT was 174.5 U/L.The liver function returned to the normal with HBV-DNA negative after larnivudine treatment in the 8 cases.HBV-DNA,however,reappeared in 5 eases without any dis- continuation of lamivudine.The creatinine level remained normal without any severe drug side effects in 28 patients during lamivudine treatment.Conclusion Lamivudine treatment before hepatic dysfunc- tion might be a safe and effective strategy for prevention of recurrence of hepatitis B viremia in HBsAg positive renal allograft recipients.

AIM: To study effects of hyperbaric oxygen (HBO) and cyclosporin A (CsA) on the contents of active oxygens and nitric oxide (NO) in spleens of skin transplanted mice. METHODS: The donor mice BALB/C and receptor mice Cs7BL/6 were tested for skin transplantation. The HBO group mice were,treated with 99.2 % oxygen under 0.25 MPa for 1.5 hours, while CsA group mice were treated with CsA 0.5 rmg· kg- t· d- 1 by abdomen injection. After 14 days, the spleen were extracted the contents of malondialdehyde (MDA) and NO and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH - PX), catalase (CAT) and NO synthases (NOS) were determined. RESULTS: (1) Compared with the control group, the transplantation group, HBO group and CsA group have markedly increased the content of MDA and the activities of GSH - PX and CAT; Compared with the transplantation group, the CsA group have markedly increased activity of SOD and reduced activities of GSH - PX and CAT; the HBO group have markedly reduced the activity of GSH - PX and increased the activities of CAT and SOD (P < 0.01 ). (2) Compared with the control group, the transplantation group have markedly increased the content of NO and the activity of NOS; Compared with the transplantation group, the HBO group have markedly increased the activity of NOS and reduced the content of NO ( P < 0.01 ); The content of NO and the activity of NOS in CsA group was not changed significantly. CONCLUSION: In the lymphocytes of the transplantation group, the peroxidation is intensified, and the content of NO and the activity of NOS increased. HBO and CsA may activate the systems of oxidation/antioxidation and NO/NOS in spleen, which may be related to their mechanism of inhibition rejection.

Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-kappaB and the expression of chemokine MOB-1, MCP-lgenes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTC in vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-kappaB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10(-3) mol/L, 10(-4) mol/L carbachol) could induce a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10(-3) mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-kappaB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10(-3) mol/L carbachol) in vitro for 30 min. Either M3 cholinergic receptor antagonist (10(-5) mol/L atropine) or NF-kappaB inhibitor (10(-2) mol/L PDTC) could obviously inhibit the activation of NF-kappaB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-lgenes in pancreatic acinar cells in vitro through the activation of NF-kappaB.

AIM: To investigate the effect of mydriasis optometry combined with amblyopia treatment on refraction and amblyopia changes in children with mixed astigmatism and amblyopia.METHODS: Totally 163 children (289 eyes) of mixed astigmatism and amblyopia from January 2010 to May 2011 were treated.All of the patients received mydriatic optometry and spectaculars with amblyopia therapy and were followed up for 5a to observe amblyopia efficacy and refractive status changes.RESULTS: With 5a, main diameter diopter at distant vision decreased year by year, average decline in the first year was 0.55DS, 0.56DS in the second year, 0.72DS in the third year, 0.95DS in the fourth year, 1.89DS in the fifth year.The spherical equivalent changed from 1.12DS at distant to 0.78DS at near.The corrected visual acuity of all the patients at first visit was 0.2-0.8 with varying degrees amblyopia.After a 5-year treatment, it was effective in 268 eyes (92.7%), in which 165 eyes (57.1%) improved, 103 eyes (35.6%) cured, the results was better as the period of spectaculars wearing was longer.There was 36 eyes (37.5%) improved and 60 eyes (62.5%) cured in 1.50-2.50DC group;118 eyes (74.2%) improved, 41 eyes (25.8%) cured in 2.50-3.50DC group;11 eyes (32.3%) improved, 2 eyes (5.9%) cured, 21 eyes (61.8%) useless in >3.50DC Group.The differences of efficacy among the groups were significant (all P<0.05).CONCLUSION: Appropriate spectaculars is the basic for amblyopia treatment.It is effective for most children with mixed astigmatism and amblyopia to take mydriasis optometry and amblyopia treatment.

OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.

METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.

RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.

CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.

Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection

Objective:To observe the effect of warm-unblocking acupuncture plus fluticasone propionate nasal spray on the pulmonary ventilation, level of interferon-γ (IFN-γ) and sleep quality in patients with allergic rhinitis (AR). Methods: A total of 112 AR patients were enrolled between January 2013 and August 2018 and were divided into an observation group and a control group by the random number table method, with 56 cases in each group. Patients in the observation group received warm-unblocking acupuncture plus fluticasone propionate nasal spray, and patients in the control group only received fluticasone propionate nasal spray. The nasal symptom score, pulmonary function indexes, the levels of IFN-γ and interleukin (IL)-4 in serum, and sleep quality in the two groups were compared. Results: After treatment, the total effective rate in the observation group was higher than that in the control group (P＜0.05). The nasal symptom score dropped in both groups after treatment (both P＜0.05), and the score in the observation group was lower than that in the control group (P＜0.05). The pulmonary ventilation indexes all increased significantly after treatment in the observation group (all P＜0.05); the forced expiratory volume in 1 second (FEV1) to forced vital capacity (FVC) ratio (FEV1/FVC) and the forced expiratory flow at 50%, 75% and 25%-75% of the vital capacity (FEF50%, FEF75%, FEF25%-75%) increased after treatment in the control group (all P＜0.05); the pulmonary ventilation indexes were higher in the observation group than those in the control group (all P＜0.05). The level of IFN-γ increased significantly after treatment in the two groups (both P＜0.05) and the level of IL-4 dropped significantly (both P＜0.05); the observation group had a higher IFN-γ level (P＜0.05) and a lower IL-4 level (P＜0.05) compared with the control group. Regarding the Pittsburgh sleep quality index (PSQI), the scores of subjective sleep quality, habitual sleep efficiency and sleep disturbances and the general PSQI score decreased significantly after treatment in both groups (all P＜0.05), and the scores in the observation group were significantly lower than those in the control group (all P＜0.05). Conclusion: Warm-unblocking acupuncture plus fluticasone propionate nasal spray can effectively control the clinical symptoms and improve pulmonary function in the treatment of AR; this approach can regulate the levels of IFN-γ and IL-4 towards the normal range in AR patients; it can also improve patient’s sleep quality. This method can produce more significant efficacy than fluticasone propionate nasal spray used alone.

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.

Twelve compounds were isolated from the venom of Bufo bufo gargarizans. On the basis of their physical and chemical properties and spectral data, their structures were identified as resibufagenin (1), bufotalin (2), desacetylcinobufagin (3), 19-oxodesacetylcinobufotalin (4), cinobufotalin (5), 1beta-hydroxylbufalin (6), 12alpha-hydroxybufalin (7), bufotalinin (8), Hellebrigenin (9), telocinobufagin (10), hellebrigenol (11) and cinobufagin-3-hemisuberate methyl ester (12), respectively. Compounds 7 and 12 are new natural products.
Animals ; Bufanolides ; chemistry ; Bufo bufo ; Medicine, Chinese Traditional ; Molecular Structure ; Venoms ; chemistry