Study on the clinical comprehensive evaluation of blood lipid-regulating drugs in five provinces and regions in Northwest China / 中国药房

OBJECTIVE To explore standardized evaluation process for clinical comprehensive evaluation of blood lipid- regulating drugs and perform rapid assessment of clinical comprehensive evaluation of blood lipid-regulating drugs with different mechanisms so as to provide reference for the drug catalogue selection and rational drug use of medical institutions. METHODS Referring to guidelines and consensus such as the guideline for the management of comprehensive clinical evaluation of drugs， the methods such as literature research， expert interviews， and Delphi expert consultation were used to establish a multi-dimensional and multi-criteria clinical comprehensive evaluation index system and quantitative scoring table for blood lipid-regulating drugs around the two main lines of technical evaluation and policy evaluation. Then 13 blood lipid-regulating drugs with different mechanisms in 21 third-grade class-A medical institutions from five provinces and regions of Northwest China were scored from both technical and policy dimensions to form a comprehensive evaluation result. RESULTS The clinical comprehensive evaluation index system and corresponding rapid evaluation quantitative scoring table were constructed for blood lipid-regulating drugs in the five northwest provinces and regions. The technicalevaluation section included 6 primary indicators， 13 secondary indicators， and 34 tertiary indicators， totaling 110 points. The policy evaluation section included 4 primary indicators and 6 secondary indicators， with a total score of 40 points （30 points for some drugs） and a total score of 150 points （or 140 points）. The scoring results showed that the highest score was atorvastatin， followed by rosuvastatin and simvastatin. CONCLUSIONS Statins are still the cornerstone of drug therapy for patients with dyslipidemia； the rapid evaluation quantitative scoring table constructed in this study is comprehensive， systematic and operable. The evaluation process in this study can provide empirical references for other groups to exploring the standardized path and quality control mechanism of clinical comprehensive evaluation of drugs.

Effects of electroacupuncture on rats with cognitive impairment: An iTRAQ-based proteomics analysis / 中西医结合学报

OBJECTIVE@#The study explores the effects of electroacupuncture (EA) at the governing vessel (GV) on proteomic changes in the hippocampus of rats with cognitive impairment.@*METHODS@#Healthy male rats were randomly divided into 3 groups: sham, model and EA. Cognitive impairment was induced by left middle cerebral artery occlusion in the model and EA groups. Rats in the EA group were treated with EA at Shenting (GV24) and Baihui (GV20) for 7 d. Neurological deficit was scored using the Longa scale, the learning and memory ability was detected using the Morris water maze (MWM) test, and the proteomic profiling in the hippocampus was analyzed using protein-labeling technology based on the isobaric tag for relative and absolute quantitation (iTRAQ). The Western blot (WB) analysis was used to detect the proteins and validate the results of iTRAQ.@*RESULTS@#Compared with the model group, the neurological deficit score was significantly reduced, and the escape latency in the MWM test was significantly shortened, while the number of platform crossings increased in the EA group. A total of 2872 proteins were identified by iTRAQ. Differentially expressed proteins (DEPs) were identified between different groups: 92 proteins were upregulated and 103 were downregulated in the model group compared with the sham group, while 142 proteins were upregulated and 126 were downregulated in the EA group compared with the model group. Most of the DEPs were involved in oxidative phosphorylation, glycolipid metabolism and synaptic transmission. Furthermore, we also verified 4 DEPs using WB technology. Although the WB results were not exactly the same as the iTRAQ results, the expression trends of the DEPs were consistent. The upregulation of heat-shock protein β1 (Hspb1) was the highest in the EA group compared to the model group.@*CONCLUSION@#EA can effect proteomic changes in the hippocampus of rats with cognitive impairment. Hspb1 may be involved in the molecular mechanism by which acupuncture improves cognitive impairment.

Wide Pulse Width Electroacupuncture Ameliorates Denervation-Induced Skeletal Muscle Atrophy in Rats via IGF-1/PI3K/Akt Pathway / 中国结合医学杂志

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.

Comparison of the efficacy of anti-vascular endothelial growth factor drugs with or without retinal laser photocoagulation for macular edema secondary to retinal vein occlusion: a systematic review / 中华眼底病杂志

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.

Effects of TAT-SOD at Acupoints on Essential Hypertension by Monitoring Meridians Electrical Potential / 中国结合医学杂志

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.

Analysis of efficiency of sacral neuromodulation in the treatment of non-neurogenic and non-obstructive dysuria in elderly patients / 中华老年医学杂志

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.

Analysis on the specificity of the relevant points along the governor vessel in chronic gastritis based on tenderness reaction and tenderness threshold / 中国针灸

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.

Analysis of efficiency of sacral neuromodulation in the treatment of non-neurogenic and non-obstructive dysuria in elderly patients / 中华老年医学杂志

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.

Evaluation of curative effect of kidney-sparing surgery in elderly patients with upper urinary tract urothelial tumors / 中华老年医学杂志

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.

Role of mitochondrial permeability transition pore in mediating the inhibitory effect of gastrodin on oxidative stress in cardiac myocytes / 南方医科大学学报

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.

Optimization of Strategy for Image Guided Radiotherapy in Tomotherapy / 中山大学学报(医学科学版)

[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.

Survival Analysis of Hepatocellular Carcinoma: A Comparison Between Young Patients and Aged Patients / 中华医学杂志(英文版)

<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>

The influence of therapeutic drug monitoring compliance on efficiency and safety about valproic acid in epileptic children / 中国药学杂志

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.

Role of vascular cell adhesion molecule-1 in the mouse model of hepatic ischemia/reperfusion and the hematogenic metastasis / 中国医学科学院学报

<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>

Expression analyses of BcUGT3 and BcUGT6, and their in vitro expression in Escherichia coli / 中国中药杂志

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.

Induction of hairy roots and plantlet regeneration of Bupleurum chinense DC / 药学学报

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.

Cloning, sequence analysis, and prokaryotic expression vector construction of glycosyltransferase gene BcUGT8 from Bupleurum chinense / 中草药

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.

Expression analysis of glycosyltransferase BcUGT1 from Bupleurum chinense DC. and its expression in E. coli and the target protein purification / 药学学报

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.

Studies on high efficient plant regeneration system from anther callus of Bupleurum chinense / 中国中药杂志

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.

Effects of cucurmosin on the cell proliferation and apoptosis in human pancreatic PANC-1 cells / 中国中西医结合杂志

<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>