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1.
Article in English | WPRIM | ID: wpr-880572

ABSTRACT

OBJECTIVE@#To evaluate the effect of the pulse width of electroacupuncture (EA) in the treatment of denervation-induced skeletal muscle atrophy in rats and examine the role of insulin-like growth factor 1 (IGF-1)/phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway during EA.@*METHODS@#Sciatic nerve functional index (SFI), muscle wet weight and the cross-sectional area (CSA) of the gastrocnemius muscle were analyzed after treatment in model rats with EA of various pulse widths (0.5, 50, 100 and 200 ms). The apoptosis index (AI) and paired box (PAX)3 and PAX7 protein expression were also determined. Further, the mRNA and protein expressions of components of IGF-1/PI3K/Akt pathway and their downstream targets were determined, along with the inhibiting effect of the pathway with a PI3-specific inhibitor.@*RESULTS@#EA with a pulse width of 200 ms was found to have the best effect with regard to increasing SFI, CSA and muscle weight, decreasing AI, and increasing the expression of PAX3 and PAX7. The IGF-1/PI3K/Akt pathway was found to be activated by denervation, although the downstream forkhead box O (FoxO) pathway was not suppressed by its activation. The PI3K/Akt pathway and its downstream molecule mammalian target of rapamycin (mTOR) were up-regulated further by EA to promote muscle protein synthesis. Meanwhile, the expressions of downstream FoxO and F-box protein 32 (ATROGIN-1) were down-regulated to reduce protein degradation.@*CONCLUSIONS@#EA with 200-ms pulse width was found to have a more significant effect than 0.5-ms EA. The positive effects of EA disappeared after inhibition of the PI3K/Akt pathway.

2.
Article in Chinese | WPRIM | ID: wpr-912396

ABSTRACT

Objective:To evaluate the efficacy of intravitreal injection of anti-vascular endothelial growth factor (VEGF) drugs combined with retinal laser photocoagulation and anti-VEGF drugs alone in the treatment of retinal vein occlusion (RVO) combined with macular edema (ME).Methods:A evidence-based medicine study. Retinal vein occlusion (obstruction), macular edema, anti-vascular endothelial growth factor, bevacizumab, ranibizumab, conbercept, aflibercept, and retinal laser photocoagulation were the Chinese and English search terms. Related literature was searched in China National Knowledge Infrastructure, Wanfang, Weipu, PubMed, Embase, Cochrane Library. RVO combined with ME was selected as the research object, and the treatment plan was a clinical randomized controlled study comparing intravitreal injection of anti-VEGF drugs combined with laser photocoagulation and anti-VEGF drugs alone. The search time range was from January 2011 to February 2021. Repeated, incomplete or irrelevant literature, case reports and review literature were excluded. Review Manager 5.4 statistical software was used to conduct a meta-analysis of the literature. The weighted mean difference (WMD) and 95% confidence interval ( CI) were selected as the estimated value of effect size, and the fixed effect model was used for analysis. The evaluation indicators were best corrected visual acuity (BCVA), center macular thickness (CMT), and the number of injections. Results:According to the search strategy, 461 articles were initially retrieved, and 21 articles were finally included for meta-analysis. A total of 1156 patients were enrolled, of which 576 were treated with anti-VEGF drugs combined with laser photocoagulation (combined treatment group), and 580 were treated with anti-VEGF drugs alone (drug treatment group). Meta-analysis results showed that there was no statistically significant difference in BCVA and CMT between the drug treatment group and the combination treatment group at 6, 9, and 12 months after treatment (BCVA: WMD =-0.82, 95% CI -2.38-0.74, P=0.30; CMT: WMD=-3.12, 95% CI -17.25-11.01, P=0.67). For patients with branch retinal vein occlusion and ME, combined therapy can reduce the number of injections more effectively than drug therapy alone ( WMD=-0.80, 95% CI -1.18--0.42, Z=4.10, P <0.000 1). Conclusion:Compared with pure intravitreal injection of anti-VEGF drugs, combined retinal laser photocoagulation can not better improve BCVA and CMT in patients with RVO and ME. For patients with branch retinal vein occlusion and ME, combined retinal laser photocoagulation can effectively reduce the resistance. The number of VEGF injections.

3.
Article in English | WPRIM | ID: wpr-827090

ABSTRACT

OBJECTIVES@#To investigate the effect on essential hypertension of the topical application of TAT-Cu, Zn-superoxide dismutase (TAT-SOD) at left acupoint Zusanli (ST 36), and to observe whether the change of electrical potential difference (EPD) can be related to the change of blood pressure.@*METHODS@#Sixteen patients with essential hypertension and 16 healthy subjects were included in the study. EPD between the left acupoints of Yanglingquan (GB 34) and Qiuxu (GB 40) was firstly screened out for the EPD detection. An intracellular superoxide quenching enzyme, TAT-SOD, was topically applied to the acupoint ST 36 within an area of 1 cm once a day, and the influence on EPD was investigated. The dosage applied to TAT-SOD group (n=8) was 0.2 mL of 3000 U/mL TAT-SOD cream prepared by adding purified TAT-SOD to a vehicle cream, while placebo group (n=8) used the vehicle cream instead. The left acupoints of Yanglingquan (GB 34) and Qiuxu (GB 40) were selected for EPD measurement after comparing EPD readings between 5 acupoints on each of all 12 meridians.@*RESULTS@#EPDs between the left acupoints of GB 34 and GB 40 for 16 patients of essential hypertension and 16 healthy subjects were 44.9±6.4 and 5.6±0.9 mV, respectively. Daily application of TAT-SOD for 15 days at ST 36 of essential hypertension patients significantly decreased systolic blood pressure (SBP) and diastolic blood pressure (DBP) of 179.6 and 81.5 mm Hg to 153.1 and 74.1 mm Hg, respectively. Responding to the change in blood pressure, EPD between the left acupoints of GB 34 and GB 40 also declined from 44.4 to 22.8 mV with the same trend. No change was observed with SBP, DBP and EPD between the left acupoints of GB 34 and GB 40 with the daily application of the placebo cream.@*CONCLUSION@#Enzymatic scavenging of the intracellular superoxide at ST 36 proved to be effective in decreasing SBP and DBP. The results reconfirm the involvement of superoxide anions and its transportation along the meridians, and demonstrate that EPD between acupoints may be an indicator to reflect its functioning status. Moreover, preliminary results suggest a close correlation between EPD and blood pressure readings, implying a possibility of using EPD as a sensitive parameter for blood pressure and to monitor the effect of antihypertensive treatment.

4.
Article in Chinese | WPRIM | ID: wpr-775857

ABSTRACT

OBJECTIVE@#To explore the specificity of the relevant points along the governor vessel in chronic gastritis through the synchronous observation of the tenderness reaction and tenderness threshold on the back along the governor vessel in the volunteers with chronic gastritis and make the comparison with healthy volunteers.@*METHODS@#A chronic gastritis group and a healthy volunteer group were prepared, 30 cases in each one. Using synchronous comparison, the tenderness reaction at the point inferior to each of the spinous processes from T to L along the governor vessel was observed. The mechanical pressure pain threshold was determined.@*RESULTS@#Regarding the tenderness reaction at the point inferior to each spinous process on the back along the governor vessel, the occurrence rate was different significantly between the chronic gastritis group and the healthy volunteer group (0.05).@*CONCLUSION@#The point inferior to the spoinous process of T on the governor vessel, Shendao (GV 11, T) and Lingtai (GV 10, T) are the points with the high specificity in chronic gastritis, which provides the reference to clinical treatment with acupuncture and moxibustion. The occurrence rate of high tenderness reaction and the low tenderness threshold may be the important manifestation of the point specificity on the back along the governor vessel in chronic gastritis.


Subject(s)
Acupuncture Points , Acupuncture Therapy , Gastritis , Therapeutics , Humans , Moxibustion , Pressure , Sensitivity and Specificity
5.
Chinese Journal of Geriatrics ; (12): 893-896, 2019.
Article in Chinese | WPRIM | ID: wpr-755437

ABSTRACT

Objective To evaluate the value of kidney-sparing surgery in the treatment of upper urinary tract urothelial tumors by retrospectively analyzing and comparing the clinical data of elderly patients with upper urinary tract urothelial tumors between patients undergoing kidney-sparing surgery and nephroureterectomy in our hospital during the same period.Methods Thirty elderly patients with upper urinary tract urothelial tumors were treated with kidney-sparing surgery(ureteroscopy,segmental ureteral resection)and followed up from April 2004 to July 2017.One hundred and fortynine patients who underwent nephroureterectomy during the same period were selected as the control group.The survival rate and local recurrence rate were compared between the two groups.Results In the kidney-sparing group,23 cases were pathologically diagnosed as tumors,3 cases as inflammatory polyps and 4 cases without pathological findings.There were no statistically significant differences between kidney-sparing surgery and nephroureterectomy in bladder recurrence[4 cases(13.3%)vs.24 cases(16.1%),x2 =0.011,P =0.915],pelvic orthotopic recurrence [3 cases (10.0%) vs.6 cases (4.0%),x2 =0.825,P=0.364],cancer-specific death rate[2 cases(6.7%)vs.16 cases(10.7%),x2=0.118,P=0.731]and overall survival rate[4 cases(13.3%)vs.22 cases(14.8%),x2 =0.410,P =0.550].There were no significant differences between kidney-sparing surgery versus nephroureterectomy among postoperative 1-year,5-year and 10-year in the overall survival rates (100.0 %vs.95.1%、85.9 % vs.84.1%、80.5 % vs.60.8 %,P =0.156),tumor-specific survival rates (100.0 % vs.95.1%,85.7 % vs.87.6 %,85.7 % vs.76.8 %,P =0.380) and progression-free survival rates(100.0%vs.100.0%,90.9%vs.96.0%,90.9% vs.79.7%,P =0.680).Conclusions Elderly patients have poor physical conditions and poor tolerance to radical surgery with a significant damage on postoperative renal function.Kidney-sparing surgery(ureteroscopy,segmental ureteral resection) as the treatment of upper urinary tract tumors can minimize the risk of surgery and optimize the quality of life.The individual assessment based on individual conditions,postoperative complications,and the recovery of renal function should be conducted and the operation with the greatest benefit should be adopted.

6.
Chinese Journal of Geriatrics ; (12): 1273-1277, 2019.
Article in Chinese | WPRIM | ID: wpr-824551

ABSTRACT

Objective To evaluate the effectiveness and safety of sacral neuromodulation(SNM) in the treatment of non-neurogenic and non-obstructive dysuria in elderly patients.Methods Six elderly patients with non-neurogenic and non-obstructive(2 males and 4 females)treated with SNM were recruited from multiple medical centers across the country in this retrospective study from January 2012 to December 2016.All patients received two or more conservative treatments including behavior therapy,oral therapy,etc.,with no good effect.Data of the average residual urine volume,average urine volume,maximum urine volume,average urinary frequency and quality of life score before operation,after implantation of stage Ⅰ tined lead,and at the end of follow-up after implantation of stage Ⅱ implanted pulse generator(IPG) were compared.Results The average age in the 6 patients was 69.5 (65 ~ 79) years.The results of SNM treatment showed that the objective findings and/or subjective symptoms in all patients were significantly improved in more than 50% of indexes before treatment.All patients chose embedding of IPG at the end of stage Ⅰ,with 100.0% of the conversion rate from stage Ⅰ to stage Ⅱ.The curative effect was stable in general.At stages of baseline,personal experience and end of follow-up respectively,the average residual urine volumes were (161.7±110.2)ml,(70.0±52.2) ml,and(50.0±44.7)ml,the average urine volume were(105.0± 74.0)ml,(163.3 ± 93.1) ml,and (155.0 ± 92.3) ml,and the maximum urine volume were (146.7 ± 81.2) ml,(216.7 ± 93.1) ml,and (206.7 ± 112.7) ml,which showed continuously significant improvements with patients' great satisfaction.The mean duration of fellow-up in 6 cases was 17.5 months(7-57 months).No severe adverse reactions such as wound infection,electrode dislocation and hematoma were observed.Conclusions SNM treatment is safe and effective for elderly patients with non-neurogenic and non-obstructive dysuria who are refractory to other treatments.SNM has a stable medium-term curative effect and a high-transfer-rate from stage Ⅰ to stage Ⅱ,which is appropriate for non-neurogenic and non-obstructive dysuria.The duration of SNM may be positively correlated with the improvement of residual urine volume and urination frequency.

7.
Chinese Journal of Geriatrics ; (12): 1273-1277, 2019.
Article in Chinese | WPRIM | ID: wpr-801263

ABSTRACT

Objective@#To evaluate the effectiveness and safety of sacral neuromodulation(SNM)in the treatment of non-neurogenic and non-obstructive dysuria in elderly patients.@*Methods@#Six elderly patients with non-neurogenic and non-obstructive(2 males and 4 females)treated with SNM were recruited from multiple medical centers across the country in this retrospective study from January 2012 to December 2016.All patients received two or more conservative treatments including behavior therapy, oral therapy, etc., with no good effect.Data of the average residual urine volume, average urine volume, maximum urine volume, average urinary frequency and quality of life score before operation, after implantation of stage Ⅰ tined lead, and at the end of follow-up after implantation of stage Ⅱ implanted pulse generator(IPG)were compared.@*Results@#The average age in the 6 patients was 69.5(65~79)years.The results of SNM treatment showed that the objective findings and/or subjective symptoms in all patients were significantly improved in more than 50% of indexes before treatment.All patients chose embedding of IPG at the end of stage Ⅰ, with 100.0% of the conversion rate from stage Ⅰ to stage Ⅱ.The curative effect was stable in general.At stages of baseline, personal experience and end of follow-up respectively, the average residual urine volumes were(161.7±110.2)ml, (70.0±52.2)ml, and(50.0±44.7)ml, the average urine volume were(105.0±74.0)ml, (163.3±93.1)ml, and(155.0±92.3)ml, and the maximum urine volume were(146.7±81.2)ml, (216.7±93.1)ml, and(206.7±112.7)ml, which showed continuously significant improvements with patients' great satisfaction.The mean duration of fellow-up in 6 cases was 17.5 months(7-57 months). No severe adverse reactions such as wound infection, electrode dislocation and hematoma were observed.@*Conclusions@#SNM treatment is safe and effective for elderly patients with non-neurogenic and non-obstructive dysuria who are refractory to other treatments.SNM has a stable medium-term curative effect and a high-transfer-rate from stage Ⅰ to stage Ⅱ, which is appropriate for non-neurogenic and non-obstructive dysuria.The duration of SNM may be positively correlated with the improvement of residual urine volume and urination frequency.

8.
Article in Chinese | WPRIM | ID: wpr-771476

ABSTRACT

OBJECTIVE@#To explore the role of mitochondrial permeability transition pore (mPTP) in mediating the protective effect of gastrodin against oxidative stress damage in H9c2 cardiac myocytes.@*METHODS@#H9c2 cardiac myocytes were treated with HO, gastrodin, gastrodin+HO, cyclosporin A (CsA), or CsA+gas+HO group. MTT assay was used to detect the survival ratio of H9c2 cells, and flow cytometry with Annexin V-FITC/PI double staining was used to analyze the early apoptosis rate after the treatments. The concentration of ATP and level of reactive oxygen species (ROS) in the cells were detected using commercial kits. The mitochondrial membrane potential of the cells was detected with laser confocal microscopy. The expression of cytochrome C was detected with Western blotting, and the activity of caspase-3 was also assessed in the cells.@*RESULTS@#Gastrodin pretreatment could prevent oxidative stress-induced reduction of mitochondrial membrane potential, and this effect was inhibited by the application of CsA. Gastrodin significantly lowered the levels of ROS and apoptosis-related factors in HO-exposed cells, and such effects were reversed by CsA. CsA significantly antagonized the protective effect of gastrodin against apoptosis in HO-exposed cells.@*CONCLUSIONS@#Gastrodin prevents oxidative stress-induced injury in H9c2 cells by inhibiting mPTP opening to reduce the cell apoptosis.


Subject(s)
Adenosine Triphosphate , Apoptosis , Benzyl Alcohols , Pharmacology , Caspase 3 , Cell Line , Cell Survival , Cyclosporine , Pharmacology , Cytochromes c , Glucosides , Pharmacology , Humans , Hydrogen Peroxide , Pharmacology , Membrane Potential, Mitochondrial , Mitochondrial Membrane Transport Proteins , Physiology , Myocytes, Cardiac , Metabolism , Oxidative Stress , Reactive Oxygen Species
9.
Article in Chinese | WPRIM | ID: wpr-668314

ABSTRACT

[Objective]To explore the different scanning frequency for the MVCT scanning in the tomotherapy , and seek the best strategy of the scanning.[Methods]Patients were treated by tomotherapy in SYSUCC from January 1, 2015 to May 15, 2016, includ?ing 88 head&neck patients, 63 chest&abdomen patients and 19 pelvis patients. All patients have had a daily MVCT scanning for po?sition verification before the treatment. The data retrieved from"the first five times of the very first week"and"the every first time of each week", were compared with"the whole course"data, respectively.[Results]There are no significant difference between"the whole course"and"the first five times of the very first week"or"the every first time of each week"for pelvis patients and chest and abdomen patients. There are significant differences between Z axis of"the whole course"and"the first five times of the very first week"only for head&neck patients.[Conclusion]It is a good scanning choice of"the first five times of the very first week"and"the every first time of each week"for the those patients with thoracic&abdominal or pelvic tumors to reduce the extra radiation dose from the MVCT. There are significant differences between Z axis of"the whole course"and"the first five times of the very first week"in the head&neck patients. Therefore,"the first five times of the very first week"cannot be used as a solid position information for daily verification. And"the every first time of each week"may be a good compromise solution.

10.
Chinese Medical Journal ; (24): 1793-1800, 2015.
Article in English | WPRIM | ID: wpr-231691

ABSTRACT

<p><b>BACKGROUND</b>To compare the clinicopathological features and prognosis between younger and aged patients with hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>We analyzed the outcome of 451 HCC patients underwent liver resection, transcatheter arterial chemoembolization and radiofrequency ablation, respectively. Then risk factors for aged and younger patients' survival were evaluated by multivariate analysis, respectively.</p><p><b>RESULTS</b>The patients who were older than 55 years old were defined as the older group. The overall survival for aged patients was significantly worse than those younger patients. The younger patients had similar liver functional reserve but more aggressive tumor factors than aged patients. Cox regression analysis showed that the elevated levels of aspartate aminotransferase (AST) (Wald χ2 = 3.963, P = 0.047, hazard ratio [HR] =1.453, 95% confidence interval [CI]: 1.006-2.098), lower albumin (Wald χ2 = 12.213, P < 0.001, HR = 1.982, 95% CI: 1.351-2.910), tumor size (Wald χ2 = 8.179, P = 0.004, HR = 1.841, 95% CI: 1.212-2.797), and higher alpha-fetoprotein level (Wald χ2 = 4.044, P = 0.044, HR = 1.465, 95% CI: 1.010-2.126) were independent prognostic factors for aged patients, while only elevated levels of AST (Wald χ2 = 14.491, P < 0.001, HR = 2.285, 95% CI: 1.493-3.496) and tumor size (Wald χ2 = 21.662, P < 0.001, HR = 2.928, 95% CI: 1.863-4.604) were independent prognostic factors for younger patients.</p><p><b>CONCLUSIONS</b>Age is a risk factor to determine the prognosis of patients with HCC. Aged patients who have good liver functional reserve are still encouraged to receive curative therapy.</p>


Subject(s)
Adult , Age Factors , Aged , Aged, 80 and over , Carcinoma, Hepatocellular , Mortality , Female , Humans , Liver Neoplasms , Mortality , Male , Middle Aged , Retrospective Studies , Survival Analysis , Young Adult
11.
Chinese Pharmaceutical Journal ; (24): 1829-1832, 2015.
Article in Chinese | WPRIM | ID: wpr-859658

ABSTRACT

OBJECTIVE: To evaluate the efficiency and safety of sodium valproate by testing the compliance of therapeutic drug monitoring (TDM) in epileptic children so as to provide a reference for rational and individual treatment. METHODS: A retrospective analysis of clinical data of children with epilepsy was performed. The patients were treated with sodium valproate or combined with other AEDs. RESULTS: A total 1 507 children were included in the study. It was found that the 49.44% of children had taken TDM only once and the children who had TDM for over 4 times merely took up 5.24%. The first time performed TDM data within therapeutic window only took up 51.63%, and 3. 58% exceeding the window as well. There were statistical difference between age, daily dosage, valproate plasma concentration and control rate among different times in TDM (P<0.001). And difference was also statistical significance between valproate plasma concentration in age groups. Comparing sodium valproate monotherapy to sodium valproate combined with other AEDs, daily dosage (P<0.001), the valproate serum concentration (P=0.001) and the rate within therapeutic window (P=0.004) were all in statistical difference. However, there was no difference in abnormal liver function rate among TDM times. CONCLUSION: Since the great difference among individuals, it need to take individual therapy based on the results of TDM and to reduce the adverse reaction so as to improve the efficiency and rational use of drugs on clinic.

12.
Article in Chinese | WPRIM | ID: wpr-319631

ABSTRACT

The tissue-specific and MeJA-induced transcriptional levels of BcUGT3 and BcUGT6 in Bupleurum chinense were analyzed in the present study. The transcriptional levels of BcUGT3 in root, leaf, flower and fruit were similar and they all were higher than those in stem. The transcriptional level of BcUGT6 was the highest in leaf and the lowest in flower among in all tested tissues. With non-treated adventitious roots as control, BcUGT6's transcriptional levels were elevated to nearly 2 folds for 2 h, 8 h, 24 h, 2 d and 4 d in MeJA-treated adventitious roots of B. chinense. It showed that the transcriptional level of BcUGT6 was slightly affected by MeJA. While, BcUGT3's transcriptional levels were gradually elevated, and till 4 d after MeJA treatment, the expression level was about 7 folds than that of non-treated control. Using pET-28a (+), the expressions of two genes was investigated. Induced by IPTG, the target proteins were expressed in E. coli and then purified. All the results obtained in the present study will be helpful for follow-up bio-function analysis of BcUGT3 and BcUGT6.


Subject(s)
Acetates , Pharmacology , Bupleurum , Cell Biology , Genetics , Cell Membrane , Metabolism , Cyclopentanes , Pharmacology , Escherichia coli , Genetics , Gene Expression , Gene Expression Regulation, Plant , Hexosyltransferases , Chemistry , Genetics , Metabolism , Intracellular Space , Metabolism , Oxylipins , Pharmacology , Protein Sorting Signals , Protein Structure, Secondary , Protein Transport , Sequence Analysis , Transcription, Genetic
13.
Article in Chinese | WPRIM | ID: wpr-329809

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of ischemia/reperfusion (I/R) on tumor metastasis in a experimental mouse model of hematogenous metastasis after I/R and to quantify expression of vascular cell adhesion molecule-1 (VCAM-1) during I/R.</p><p><b>METHODS</b>An experimental mouse model of metastasis after partial hepatic I/R was designed to determine the effects of I/R on tumor metastasis to liver. Tumor loads were valued 14 days after operation. In addition, the expressions of alanine transaminase (ALT), aspartate transaminase (AST), and VCAM-1 were detected.</p><p><b>RESULTS</b>Two hours after hepatic reperfusion, ALT and AST levels in ischemia 45-minute group and ischemia 30-minute group were significantly higher than in the sham group (all P < 0.05). Also, the changes of ALT and AST were more obvious in the ischemia 45-minute group than in ischemia 30-minute group (all P < 0.05). In the sham group, both ALT and AST slightly and transiently increased. ALT and AST in the ischemia 45-minute group and ischemia 30-minute group at 8 hours were both significantly higher than those at 2 hours reperfusion (P<0.05). The tumor load (valued by hepatic replacement area) and the expression of VCAM-1 in ischemic lobe were significantly larger in the ischemia 45-minute group than in the ischemia 30-minute group and sham group (P = 0.013, P = 0.007). However, there was no statistical difference on tumor load between the right lobe of sham operated mice and the right lobe (nonischemic lobes) of mice subjected to I/R (P = 0.089). Mouse survivals were significantly longer in the sham group than in the ischemia 30-minute group (P = 0.041) but were not significantly different between the ischemia 45-minute group and ischemia 30-minute group (P = 0.055). VCAM-1 expression in ischemia 45-minute group was significantly higher than in ischemia 30-minute group and sham group(P = 0.003, P < 0.001), and it was positively correlated with the hepatic replacement area (r = 0.491, P = 0.045).</p><p><b>CONCLUSION</b>Hepatic I/R promotes liver hematogenic metastasis of hepatocellular carcinoma in mice and at least in part, through the induction of VCAM-1 expression.</p>


Subject(s)
Animals , Liver , Liver Neoplasms , Pathology , Male , Mice , Mice, Inbred BALB C , Neoplasm Metastasis , Reperfusion Injury , Vascular Cell Adhesion Molecule-1 , Physiology
14.
Article in Chinese | WPRIM | ID: wpr-291307

ABSTRACT

The callus of Bupleurum chinense with anthers at the stage of uninucleate was induced. After several subcultures, anther calli of B. chinense were cultured at 20 MS culture mediums with different plant hormones to differentiate into plantlets. Differentiation of callus was detected after 21 and 49 days to select the most effective medium. There were 19 culture mediums in which anther callus could differentiate into plantlets with differentiation rate range from 3% to 60% , and most less than 20%. MS + KT 0.5 mg x L(-1) + sucrose 30 g c L(-1) + phytagel 5 g x L(-1) was the best differentiation medium with the differentiation rate of 60%, followed by MS + ZT 1.0 mg x L(-1) + sucrose 30 g x L(-1) + phytagel 5 g x L(-1) with the differentiation rate of 58%. Then plantlets were transferred to rooting medium to obtain whole plant. All plantlets could root in the rooting medium of MS + sucrose 30 g x L(-1) + phytagel 5 g x L(-1) and 1/2 MS + NAA 0.5 mg x L(-1) + sucrose 30 g x L(-1) + phytagel of 5 g L(-1) with the rooting rate of 100%. As a result, the high efficient and stable plant regeneration system was established from anther callus of B. chinense.


Subject(s)
Bupleurum , Metabolism , Culture Media , Chemistry , Metabolism , Flowers , Metabolism , Plant Growth Regulators , Metabolism , Seedlings , Tissue Culture Techniques , Methods
15.
Acta Pharmaceutica Sinica ; (12): 1491-1497, 2013.
Article in Chinese | WPRIM | ID: wpr-298053

ABSTRACT

In this study, the induction of hairy roots of Bupleurum chinense DC. was explored and established after experiments at different conditions: A. rhizogenes A4 was used to infect the leaves bases of B. chinense tube seedlings. The explants were co-cultured on Phytagel-solidified media for 3 days and then, were turned into solid media, similar with the co-culture media except that bacteriostat was added. After 10 days, rootlets began to appear and after 4 to 5 weeks, rootlets can be converted into liquid shaking culture stage. Plants regeneration from hairy root was useful for the research of new germplasm production and the variety improvement breeding. In the present study, the regenerated plants were obtained. One approach was to continuously culture under light conditions the seedlings which parting off spontaneously from the hairy roots during liquid shaking culture. The other approach was to culture the callus-like tissues produced by hairy roots with the optimized regeneration media for the induction of regenerated plants. The results of present study provide a technique to induce hairy roots and plantlet regeneration of B. chinense and this technique is helpful for the researches on metabolism, especially on the Agrobacterium-mediated genetic transformation of B. chinense.


Subject(s)
Agrobacterium , Genetics , Bupleurum , Genetics , Coculture Techniques , Plant Leaves , Genetics , Plant Roots , Genetics , Plants, Genetically Modified , Genetics , Plants, Medicinal , Genetics , Regeneration , Transformation, Genetic
16.
Acta Pharmaceutica Sinica ; (12): 1345-1352, 2013.
Article in Chinese | WPRIM | ID: wpr-259471

ABSTRACT

The ORF sequence of glycosyltransferase gene BcUGT1 cloned from Bupleurum chinense DC. was analyzed and its three dimentional structure was predicted. Using qRT-PCR method, the expression characteristics of BcUGT1 after methyl jasmonate (MeJA) induction and in different plant tissues were investigated. The results showed that BcUGT1 may be involved in saikosaponin biosynthesis in B. chinense. Thereafter, the recombinant vectors of BcUGT1 were constructed for its expression in E. coli. The target protein was successfully expressed and purified. In the present study, three vectors, pRSET-A, pET-28a (+) and pET-30a (+), and three isolates of E. coli, BL21 (DE3) plysS, BL21A1 and BL21-CodonPlus (DE3)-RIPL were used under different induction conditions, such as different concentrations and during times of inducers (L-arabinose and IPTG) and different inducing temperatures. The results showed that in the condition of 0.5 or 1 mmol x L(-1) IPTG, 16 degrees C, 20 h, target protein expressed in BL21-CodonPlus (DE3)-RIPL with pET-28a (+) or pET-30a (+) as vector. Using PrepEase His-tagged protein purification kit, the target protein was purified. The present work will be helpful for follow-up bio-function analysis of BcUGT1.


Subject(s)
Amino Acid Sequence , Base Sequence , Bupleurum , Chemistry , Cloning, Molecular , DNA, Complementary , Genetics , DNA, Plant , Genetics , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Glycosyltransferases , Genetics , Metabolism , Oleanolic Acid , Open Reading Frames , Genetics , Phylogeny , Plants, Medicinal , Chemistry , Protein Structure, Secondary , Recombinant Fusion Proteins , Genetics , Metabolism , Saponins
17.
Article in Chinese | WPRIM | ID: wpr-855166

ABSTRACT

Objective: To clone the full-length cDNA of the uridine diphosphate glycosyltransferase (UGT) gene in Bupleurum chinense (BcUGT8), which may be involved with the saikosaponin biosynthesis, and to construct the prokaryotic expression vector. The work will provide the foundation for its further function verification by in vitro expression and activity analysis of the purified protein. Methods: RACE and LD-PCR were used to clone the full-length cDNA of BcUGT8, on the basis of its partial cDNA sequence obtained from our previous high-flux sequencing by Roche (454) GS FLX system. The open reading form (ORF) was PCR cloned using primers with corresponding restriction enzymes cutting sites. The PCR products were digested with corresponding restriction enzymes and then were inserted in expression vector pET-28a (+) to construct the recombinant expression vectors. Results: The full-length cDNA of UGT gene was cloned from B. chinense, and the prokaryotic expression vector was obtained. Conclusion: The full-length cDNA cloning, sequence analysis, and prokaryotic expression vector construction provide a substantial foundation for follow-up bio-function analysis of BcUGT8 through protein expression, purification, and activity analysis in vitro.

18.
Article in Chinese | WPRIM | ID: wpr-252570

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effects of cucurmosin (CUS) on the cell proliferation and apoptosis in pancreatic PANC-1 cells.</p><p><b>METHODS</b>The inhibition of CUS on the PANC-1 cell growth was observed using MTT assay. The inhibition ratio of CUS on the pancreatic orthotopic transplantation was in vivo observed in the NOD/SCID mouse model. The changes of microstructure of the apoptosis-inducing effect of CUS on PANC-1 was observed under electron microscope. The cell cycle and apoptosis after CUS intervention was detected using flow cytometry. The Caspase-3 activity after CUS treatment was detected using enzyme linked immunospecific assay (ELISA).</p><p><b>RESULTS</b>Treatment with CUS at the dose of 0.125, 0.25, and 0.5 mg/kg inhibited the growth of pancreatic carcinoma PANC-1 xenografs with the ratio of 45.2%, 50.0%, and 59.7%, respectively (P < 0.05). After exposure to 10 microg/mL CUS for 24 h, most cells presented typical morphologic changes of apoptosis such as chromatin condensation and shrunken nucleus. The apoptotic cells increased. Some nuclear shrinkage and fragmentation, as well as the apoptotic body were observed when cells were exposed to CUS for 72 h. Being exposed to 0, 2.5, 10.0, and 40.0 microg/mL of the CUS for 72 h, the percentage of G0/G1 phase cells was 46.56% +/- 5.08%, 53.33% +/- 5.05%, 67.50% +/- 6.50%, and 77.00% +/- 6.73%, respectively (P < 0.05). The apoptosis ratio was 2.50% +/- 0.13%, 8.30% +/- 1.23%, 23.40% +/- 2.45%, and 48.50% +/- 3.65% shown by Annexin V/PI (P < 0.05). The Caspase-3 activity (unit) was 0.009 +/- 0.002, 0.011 +/- 0.003, 0.035 +/- 0.009, and 0.065 +/- 0.009, respectively (P < 0.05). These data showed that CUS induced the apoptosis of PANC-1 cells in a dose-dependent maner. Being exposed to 40.0 microg/mL of the CUS for 24, 48, and 72 h, the percentage of G0/ G1 phase cells was 56.60% +/- 6.65%, 67.83% +/- 6.76%, and 77.00% +/- 6.73%, respectively (P < 0.05), the apoptosis ratio was 16.51% +/- 2.97%, 38.51% +/- 2.38%, and 48.50% +/- 3.65% shown by Annexin V/PI (P < 0.05). These data showed that CUS induced apoptosis of PANC-1 cells in the G0/G1 phase of the cell cycle in a time-dependent maner.</p><p><b>CONCLUSION</b>CUS significantly inhibited the growth of PANC-1 cells possibly through the G0/G1 cell cycle arrest and apoptosis.</p>


Subject(s)
Animals , Apoptosis , Cell Line, Tumor , Cell Proliferation , Cucurbita , Female , Humans , Male , Mice , Mice, Inbred NOD , Mice, SCID , Pancreatic Neoplasms , Pathology , Plant Proteins , Pharmacology
19.
Article in Chinese | WPRIM | ID: wpr-230448

ABSTRACT

<p><b>OBJECTIVE</b>To find out the regular pattern of pain reaction by pressing along the Governor Vessel at the levels of T3 to L4 on the back in patients with digestive system disease, so as to provide references for diagnosis of the disease.</p><p><b>METHODS</b>Thirty patients diagnosed with digestive system disease by gastroscopy and colonoscopy as well as 16 patients with digestive symptoms without accurate diagnosis by endoscopy checks were observed. Pressing was applied from Zhiyang (GV 9) to Yaoyangguan (GV 3), including non-acupoints along the Governor Vessel. Positive reaction was recognized since pain, soreness or distension sensation appeared in the space between the spinous process.</p><p><b>RESULTS</b>The positive ratios of 30 patients with endoscopy checks and 16 without the check accounted for 100.0%. The tender spots, usually 5 to 9 in number, were mainly focused on the relative acupoints and non-acupoints along the Governor Vessel from T6 to T10.</p><p><b>CONCLUSION</b>The pain reaction from Lingtai (GV 10) to Zhongshu (GV 7), which are at the level of T6 to T10, along the Governor Vessel is closely related with digestive system disease. And it can be regarded as function criterion of the digestive system and important reference of auxiliary diagnosis.</p>


Subject(s)
Acupuncture Points , Adult , Aged , Back , Diagnosis, Differential , Digestive System Diseases , Diagnosis , Female , Humans , Male , Meridians , Middle Aged , Pain , Pain Measurement , Sensation
20.
Chinese Medical Journal ; (24): 19-25, 2011.
Article in English | WPRIM | ID: wpr-241537

ABSTRACT

<p><b>BACKGROUND</b>Molecular targeted drugs is now widely used in non-small cell lung cancer (NSCLC) clinical treatment. Icotinib hydrochloride is a new type of oral epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (EGFR-TKIs). In this study, we examined the role of EGFR, K-RAS, B-RAF somatic mutations and EGFR mRNA expression in tumor specimens from advanced NSCLC patients as predicators of the efficacy of icotinib hydrochloride.</p><p><b>METHODS</b>We analyzed tumor paraffin-embedded specimens, which were obtained from 14 of 40 patients with advanced NSCLC who enrolled in the stage I clinical trial of icotinib hydrochloride. Somatic mutations were evaluated by mutant-enriched liquidchip (MEL) technology, and EGFR mRNA expression was measured by branched DNA liquidchip (MBL) technology.</p><p><b>RESULTS</b>In the 14 specimens, seven patients showed EGFR mutations, exon 19 deletion (3/7) and exon 21 point mutation (4/7); and two patients showed K-RAS mutation. No mutations in EGFR exon 20 or B-RAF were detected. In patients with EGFR mutation, one patient developed progress disease (PD), three patients had stable disease (SD), two patients had partial responses (PR) and one patient had a complete response (CR). In patients with wild-type EGFR, four patients had PD, three patients acquired SD, and none had PR/CR (P = 0.0407). EGFR mutations were associated with better progress-free survival (PFS) (141 days vs. 61 days) but without a statistically significant difference (P = 0.8597), and median overall survival (OS) (≥ 449 days vs. 140 days). EGFR mRNA expression levels were evaluated (three high, eight moderate, one low, and two that can not be measured due to insufficient tumor tissue) and no statistically significant relationships was observed with response, PFS or OS.</p><p><b>CONCLUSIONS</b>The EGFR mutation rate was consistent with that reported in the Asian population, so the MEL technology is reliable for measuring EGFR mutation with high throughput and rapidity. EGFR exon 19 deletions and exon 21 point mutation are predictive biomarkers for response to icotinib hydrochloride as second line treatment or above.</p>


Subject(s)
Adult , Aged , Antineoplastic Agents , Therapeutic Uses , Carcinoma, Non-Small-Cell Lung , Drug Therapy , Genetics , Crown Ethers , Therapeutic Uses , Exons , Genetics , Female , Humans , Male , Middle Aged , Mutation , Proto-Oncogene Proteins B-raf , Genetics , Proto-Oncogene Proteins p21(ras) , Genetics , Quinazolines , Therapeutic Uses , RNA, Messenger , Genetics , ErbB Receptors , Genetics
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