The combination of high-speed countercurrent chromatography and rapid bioactive assay is a desirable method in the screening of bioactive components from CMM and natural products.This review focused on the application status of high-speed countercurrent chromatography to the rapid screening of bioactive components from CMM and natural products based on our previous studies and the recent progress over it.

This study aimed at preparing the chemical reference substance of garcinol.A preparative high-speed countercurrent chromatography (HSCCC) was adopted for the isolation of garcinol from the twigs of Garcinia multifiora Champ,G.esculenta Y.H.Li and the fruits of G.xanthochymus Hook.f.ex T.Anders.The crude extracts were separated by HSCCC with two phase solvent systems composed of n-hexane-ethyl acetate-95％ ethanol-water (8∶8∶12∶4,volume ratio) using the upper stationary phase and the lower mobile phase.Under the conditions of a flow rate of 2.0 and 4.0 mL· min-1,and the apparatus rotation at 850 rpm,the column temperature at 25℃ and the detection wavelength at 254 nm,the fraction containing garcinol was purified by the Sephadex LH20 chromatography.The purity test in the extracts was determined by high-performance liquid chromatography.As a result,the contents of garcinol were 45,281 and 4080mg respectively separated from 104.765 g twigs of G.multiflora Champ,105.270 g G.esculenta Y.H.Li and 102.318 g the fruits of G.xanthochymus Hook.f.ex T.with the purity of 98.72％,98.36％ and 98.42％.Compared with the sample pretreatments and separation efficiency of the three plants,it was found that the fat-soluble extracts rapidly and efficiently extracted using n-hexane-ethyl acetate-95％ ethanol-water (5∶5∶5∶5,volume ratio) contained abundant garcinol taking fruits of G.xanthochymus Hook.f.ex T as the raw material with the combination of HSCCC and Sephadex LH-20 chromatography.In conclusion,it was indicated that the combination method is efficient with high operability and large preparation quantity.

Objective To evaluate the role of autophagy in hippocampal neurons in cognitive dysfunction caused by sevoflurane anesthesia in juvenile rats.Methods One hundred four male SpragueDawley rats, aged 7 days, weighing 10-16 g, were randomly assigned into 3 groups using a random number table: control group (group C, n =8), sevoflurane anesthesia group (group S, n =48), and sevoflurane anesthesia + rapamycin group (group SR, n =48).Group C inhaled 60％ oxygen for 6 h.S and SR groups inhaled 3.6％ sevoflurane anesthesia for 6 h, and in addition, rapamycin 2 mg/kg was injected intraperitoneally at 1 h before of sevoflurane inhalation in group SR.The equal volume of phosphate buffer solution was given in C and S groups.At 1 h before sevoflurane anesthesia (T0) , immediately after the end of anesthesia (T1) , and at 12, 24 and 48 h after the end of anesthesia (T2-4) , 8 rats were randomly selected from S and SR groups to determine the expression of autophagy-related proteins microtubule-associated protein 1 light chain 3 (LC3) Ⅰ , LC3 Ⅱ and Beclin-1 in hippocampal neurons (by Western blot).The ratio of LC3 Ⅱ/LC3 Ⅰ was calculated.Morris water maze test was performed at 5 weeks after the end of anesthesia to assess the cognitive function.The escape latency, frequency of crossing the original platform, and duration of swimming spent at the target quadrant were recorded.Results Compared with the values at T0, the expression of LC3 Ⅱ and Beclin-1 was significantly down-regulated at T1-3 , and the ratio of LC3 Ⅱ/LC3 [was decreased in group S, and the expression of LC3 Ⅱ and Beclin-1 was significantly up-regulated at T1-4, and the ratio of LC3 Ⅱ/LC3 Ⅰ was increased in group SR (P＜0.05).Compared with group C, the escape latency was significantly prolonged, the frequency of crossing the original platform was reduced, and the duration of swimming spent at the target quadrant was shortened at 3-5 days in group S (P＜0.05) , and no significant change was found in the parameters mentioned above in group SR (P＞ 0.05).Compared with group S, the escape latency was significantly shortened, the frequency of crossing the original platform was increased, and the duration of swimming spent at the target quadrant was prolonged at 3-5 days, the expression of LC3 Ⅱ and Beclin-1 was up-regulated at T1-4 , and the ratio of LC3 Ⅱ/LC3Ⅰ was increased in group SR (P＜0.05).Conclusion Weakened autophagy in hippocampal neurons is involved in cognitive dysfunction caused by sevoflurane anesthesia in the juvenile rats.

Objective:To evaluate the role of ten-eleven translocation methylcytosine dioxygenase 3 (TET3) in trigeminal ganglion in maxillofacial inflammatory pain in mice.Methods:Forty SPF healthy male C57BL/6J mice, aged 8-10 weeks, weighing 19-23 g, were divided into 5 groups ( n=8 each) using a random number table method: control group (group C), inflammatory pain group (group IP), control+ TET3-siRNA group (group C+ siTET3), inflammatory pain+ TET3-siRNA group (group IP+ siTET3) and inflammatory pain+ negative control Scrambled-siRNA group (group IP+ siNC). Normal saline or complete Freund′s adjuvant (CFA) 10 μl was injected into the temporomandibular joint of mice, respectively, and the mechanical paw withdrawal threshold (MWT) was measured at 1, 4, 8 and 12 days after injection (T 1-4). Before injection of normal saline or CFA, 0.75 μl siTET3 or siNC was injected into the trigeminal ganglion and the animals were then sacrificed and trigeminal ganglion was removed at T 2 for determination of the expression of TET3 by Western blot in C+ siTET3, IP+ siTET3 and IP+ siNC groups. Results:Compared with group C, MWT was significantly decreased at T 1-3 , the expression of TET3 in trigeminal ganglion was up-regulate in group IP ( P<0.05 or 0.01). Compared with IP and IP+ siNC groups, MWT was significantly increased at T 2, 3, and the expression of TET3 in trigeminal ganglion was down-regulate in group IP+ siTET3 ( P<0.05 or 0.01). Conclusion:TET3 in trigeminal ganglion is involved in the development of maxillofacial inflammatory pain in mice.

Objective To understand the clinical distribution characteristics of Acinetobacter baumannii in our hospital and the change situation of drug resistance rates to provide a basis for the clinical rational drug use and the nosocomial infection manage-ment.Methods The Acinetobacter baumannii strains isolated in our hospital from January 2005 to December 2013 were performed the retrospective analysis on its department distribution,specimen distribution and change of drug resistance rates.Results 964 strains of Acinetobacter baumannii were isolated during these 9 years,in which 713 strains were multi-drug resistant.The isolated strains were less during 2005 -2008,which were 30,26,22,19 strains respectively.The isolated strains began to increase during 2009-2010,which were 65,50 strains respectively.The detection rate began to enormously increase from 2011,which were 157, 229,366 strains respectively from 2011 to 2013.The top three departments of the highest isolation rates during these 9 years were ICU,neurosurgery department and respiratory department.The specimen source was always dominated by the respiratory tract specimens,followed by the secretion samples,in recent years,the detection rates of blood,urine and drainage specimens were in-creased to some extent.The drug resistance rates in 13 kinds of drugs totally showed the increasing trend,the resistance rate of par-tial drugs was decreased to some extent.Conclusion Acinetobacter baumannii easily cause nosocomial infections,which is difficult to be eliminated and has high occurrence in the departments centralized with critical patients.The respiratory infection is the main pathogenic type.Its drug resistance is serious,multi-drug resistant and pan-resistant strains have the higher proportion.Clinic should rationally use the drugs based on the drug susceptibility test results,coordinates with the infection control departments for doing disinfection and isolation well and prevent ing the outbreak of nosocomial infections.

Objective To develop and evaluate a porcine model for laparoscopic ureterovesical reimplantation (LUR) training. Methods Ten female pigs with a mean weight of 30 kg were used and the animals were placed on supine position after anesthesia. One 10 mm port and two 5 mm ports were placed after the establishment of pneumoperitoneum. The horn of uterus was used as "ureter". A model simulating the performance of LUR was then established on the mini-pigs. Four trainees per-formed the LUR procedures on the animal models during an advanced laparoscopic urology training course, following the technique criteria exactly used in LUR. The learning curve was analyzed in terms of operation time. Results The porcine model for laparoscopic training was established suc-cessfully and 4 LUR trainings was performed on each porcine. Each trainee performed 10 LURs on the models during the training course. The operation time declined from 170±10 rain initially to 90±4 rain after the training course (P<0.01). At the end of this training, all trainees could accomplish a watertight LUR procedure on the model. Conclusions The establishment of the training model is feasible. The trainees could acquire skills needed to perform LUR in vivo based on this simple model and to develop dexterity and facility in laparoscopic manipulation of needles and sutures as well. The model provides a platform for basic techniques training of the ureteral reconstruction procedures.

Objective To evaluate the role of 2B-containing NMDA receptors(NR2B)in sevoflu-rane anesthesia-induced cognitive dysfunction in aged rats.Methods Thirty-two healthy male Sprague-Dawley rats,aged 18 months,weighing 570-630 g,were divided into 4 groups(n＝8 each)using a ran-dom number table method: control group(group C),sevoflurane anesthesia group(group S),sevoflurane anesthesia plus NR2B specific inhibitor Ro 25-6981 group(group S+RO)and Ro 25-6981 group(group RO).S and S+RO groups inhaled 3％sevoflurane for 4 h.Ro 25-6981 1 mg/kg was intraperitoneally injec-ted at 15 min before inhaling sevoflurane in group S+RO.Morris water maze test was performed at 2 days af-ter the end of anesthesia to assess cognitive function.The rats were then sacrificed,and hippocampal tis-sues were obtained for determination of the expression and phosphorylation of ERK1/2 by Western blot.Results Compared with group C,the escape latency was significantly prolonged,the frequency of crossing the original platform was reduced,the time of staying at the original platform quadrant was shortened,and the phosphorylation of ERK1/2 was decreased in group S(P＜0.05),and no significant change was found in the escape latency in S+RO and RO groups(P>0.05).Compared with group S,the escape latency was significantly shortened,the frequency of crossing the original platform was increased,the time of staying at the original platform quadrant was prolonged,and the phosphorylation of ERK1/2 was increased in group S+RO(P＜0.05).There was no significant difference in ERK1/2 expression among the four groups(P>0.05).Conclusion The mechanism by which sevoflurane anesthesia induces cognitive dysfunction is re-lated to up-regulating the expression of NR2B and inhibiting the activity of ERK1/2 in aged rats.

Objective: To evaluate the role of 2B-containing NMDA receptors (NR2B) in sevoflurane anesthesia-induced cognitive dysfunction in aged rats. Methods: Thirty-two healthy male Sprague-Dawley rats, aged 18 months, weighing 570-630 g, were divided into 4 groups (n=8 each) using a random number table method: control group (group C), sevoflurane anesthesia group (group S), sevoflurane anesthesia plus NR2B specific inhibitor Ro 25-6981 group (group S+ RO) and Ro 25-6981 group (group RO). S and S+ RO groups inhaled 3% sevoflurane for 4 h. Ro 25-6981 1 mg/kg was intraperitoneally injected at 15 min before inhaling sevoflurane in group S+ RO.Morris water maze test was performed at 2 days after the end of anesthesia to assess cognitive function.The rats were then sacrificed, and hippocampal tissues were obtained for determination of the expression and phosphorylation of ERK1/2 by Western blot. Results: Compared with group C, the escape latency was significantly prolonged, the frequency of crossing the original platform was reduced, the time of staying at the original platform quadrant was shortened, and the phosphorylation of ERK1/2 was decreased in group S (P<0.05), and no significant change was found in the escape latency in S+ RO and RO groups (P>0.05). Compared with group S, the escape latency was significantly shortened, the frequency of crossing the original platform was increased, the time of staying at the original platform quadrant was prolonged, and the phosphorylation of ERK1/2 was increased in group S+ RO(P<0.05). There was no significant difference in ERK1/2 expression among the four groups (P>0.05). Conclusion The mechanism by which sevoflurane anesthesia induces cognitive dysfunction is related to up-regulating the expression of NR2B and inhibiting the activity of ERK1/2 in aged rats.

Objective:To explore the efficacy of flow diversion combined with coil embolization in treatment of intracranial aneurysms.Methods:The clinical data of 110 patients with intracranial aneurysms treated by flow diversion in our hospital from April 2015 to September 2019 were retrospectively analyzed. In these patients, 48 were treated by flow diversion combined with coil embolization and 62 were treated by flow diversion alone; the efficacy and safety of patients from the two groups were compared.Results:Blood flow diversion was successfully implanted into all 110 patients, with technical success rate of 100%. Immediate complete occlusion rate in the flow diversion combined with coil embolization group (16.7%) was significantly higher than that in the flow diversion group (1.6%, P<0.05). There was no significant difference in the incidence of perioperative complications between flow diversion combined with coil embolization group and flow diversion group (4.17% vs. 4.84%, P>0.05). During the mean follow-up of 6.72±3.80 months, modified Rankin scale (mRS) scores of patients in the flow diversion combined with coil embolization group were all 0; one patient had mRS score of 1, one patients had mRS scores of 2, and 60 patients had mRS score of 0 in patients from the flow diversion group; no significant difference was noted between the two groups ( P>0.05). Patients in the flow diversion combined with coil embolization group had significantly higher rate of complete aneurysm occlusion than those in the flow diversion group (88.3% vs. 66.1%, P<0.05). Conclusions:Flow diversion is an effective and safe strategy in treatment of intracranial aneurysms. Flow diversion combined with coil embolization can effectively promote early healing of aneurysms in selective patients.

Objective:To explore the clinical efficacy and safety of different surgical procedures of Mayo level Ⅳ inferior vena cava tumor thrombus(IVC-TT).Methods:The clinical and pathological data of 36 patients with Mayo level Ⅳ tumor thrombus were collected in three large clinical centers in China, including 18 cases in PLA General Hospital, 7 cases in Nanfang Hospital, and 11 cases in Renji Hospital. There were 25 males and 11 females.The median age was 56.5 years (53-67 years old). The average body mass index was 24.18±2.55 kg/m 2. The average diameter of renal tumors was 8.24±3.25 cm. The average length of inferior vena cava tumor thrombus was 12.89±2.50 cm. Mayo level Ⅳ tumor thrombus were divided into level Ⅳa and level Ⅳb (301 classification) based on the criterion of whether the proximal end of the thrombus has invaded the right atrium. Among them, level Ⅳa patients underwent robot-assisted inferior vena cava thrombectomy without cardiopulmonary bypass(CPB-free group, 6 cases). Level Ⅳb patients underwent robot-assisted inferior vena cava thrombectomy with cardiopulmonary bypass(CPB group, 12 cases) or cardiopulmonary bypass with deep hypothermic circulatory arrest assisted inferior vena cava thrombectomy(CPB/DHCA group, 18 cases). The baseline data of the three groups of patients were comparable. The perioperative results and long-term survival data after surgery were compared with different surgical methods for grade Ⅳcancer thrombosis. Results:All operations were successfully completed. Compared with the CPB group, the CPB-free group had a shorter first portal blocking time[17.5(15-36)min vs. 36.5(12-102)min, P=0.044], less intraoperative bleeding [2 350(1 000-3 000)ml vs. 3 500 (1 500-12 000)ml, P=0.043] and a lower allogeneic blood transfusion [1 250(500-2 000)ml vs. 2 185(700-5 800)ml, P=0.049]. Compared with the CPB/DHCA group, the CPB-free group had an advantage in reducing intraoperative allogeneic blood transfusion [1 250(500-2 000)ml vs. 2 700(1 200-10 000)ml, P=0.003]. There were no significant differences between groups in terms of duration of surgery and postoperative hospital stay. Among the 36 patients in this group, 23(64%) developed major complications (level Ⅲ or above), including 9 (25%) grade Ⅲ, 12 (33%) grade Ⅳ, and 2 (6%) grade Ⅴ. The CPB-free group had a relatively low complication rate of grade Ⅳ or above [ 17% (1/6) vs.42% (5/12) vs.44% (8/18)]. There were no statistical differences in median progression-free survival (16.4 vs.12.3 vs.18.0 months, P=0.695) and overall survival (30.1 vs.30.2 vs.37.7 months, P=0.674) between the groups. Conclusions:Robot-assisted inferior vena cava thrombectomy without cardiopulmonary bypass has the advantages of short ischemia time of organs, less intraoperative bleeding, and low incidence of major complications, which can be used as a safe and feasible surgical strategy for selected level Ⅳ tumor thrombus.