OBJECTIVE:To optimize the extraction technology of Tibetan medicine Pedicularis kansuensis and compare con-tent of verbascoside and isoverbascoside differences in P. kansuensis from various habitats. METHODS:Using verbascoside and iso-verbascoside and dry paste yield as comprehensive evaluation indexes,single factor test and orthogonal test were used to investigate the extraction solvent,solvent dosage,extraction time and times to optimize extraction technology,and the verification test was conducted. Contents of the 2 constituents verbascoside and isoverbascoside in P. kansuensis from Gansu,Qinghai and Sichuan were compared. RESULTS:The optimal extraction technology was as follows as 8-fold 50% ethanol,extraction for 3 times,90 min each time. The verification results showed that the average contents of verbascoside and isoverbascoside were 3.49%(RSD=1.28%,n=3),1.26%(RSD=1.32%,n=3),and average dry paste yields were 37.99%(RSD=1.97%,n=3). The contents of verbascoside and isoverbascoside in P. kansuensis from Qinghai were relatively higher. CONCLUSIONS:Optimized extraction tech-nology is reasonable,stable,feasible;the contents of index constituents in P. kansuensis from different habitats have certain differ-ences. The study can provide scientific evidence for the development and utilization of extraction,and the in-depth study of quality evaluation for medicinal material.

Objective To observe the impact of mild hypothermia (MH)on the reactive oxygen species (ROS)and expression of cacpase-3mRNA and light chain 3 (LC3,a subunit of immunoglobulin)in hippocampus nerve cells of rats after cardiopulmonary resuscitation (CPR).Methods A total of 65 healthy male Sprague Dawley (SD)adult rats were randomly (random number)divided into 2 groups:blank control group (n =5)and CPR group (n =60).Cardiac arrest (CA)was induced in rats of CPR group by asphyxia.The survival rats after CPR were randomly (random number)divided into 2 groups:normothermia CPR group (NT)and hypothermia CPR group (HT).Homeothermia of 37 ℃ was maintained in NT group after restoration of spontaneous circulation (ROSC),and hypothermal intervention to 32 ℃ was carried out in HT group for 4 hours immediately after ROSC.Both NT group and HT group were then randomly divided to 2 subgroups 12 hours and 24 hours after ROSC (NT-12,NT-24,HT-12,HT-24 subgroups).During observation,the neurological deficit (NDS)of rats was scored,then the bilateral hippocampi were obtained from rats'head,and monoplast suspension of fresh hippocampus tissue was made immediately to determine the level of intracellular ROS by flow cytometry.Transmission electron microscope was used to observe the ultrastructure changes of cellular nucleus and mitochondria.Reverse transcription-polymerase chain reaction (RT-PCR)was used to determine the expression of caspase-3mRNA and Western-blotting (WB)was used to determine the level of LC3 in frozen hippocampus tissue.Measured data were analyzed with paired sample T test and One-Way ANOVA.Results Of 60 rats with CA,44 were successfully resuscitated (73%)and 33 survived until the end of the experiment (55%).The NDSs of rats in NT and HT groups were significantly reduced in comparison with BC group (F=8.107,P<0.05),while the NDSs of rats in HT-12 subgroup and HT-24 subgroup were significantly increased in comparison with NDSs of rats in NT-12 subgroup and NT-24 subgroup,respectively (t=9.692,P<0.01;t=14.374,P<0.01 ).The ROS in hippocampus nerve cells of rats in NT group and HT group were significantly increased compared to BC group (F=16.824,P<0.05 ),whereas the ROS in HT-12 and HT-24 subgroups were significantly reduced compared to ROS in NT-12 and NT-24 subgroups,respectively (t =9.836,P<0.01;t =7.499,P<0.01).The expressions of caspase-3 mRNA in hippocampus nerve cells of rats in NT and HT groups were significantly increased compared to BC group (F=24.527,P<0.05),while the expressions of caspase-3 mRNA in rats of HT-12 and HT-24 subgroups were significantly reduced compared to NT-12 and NT-24 subgroups,respectively (t =6.935,P <0.01;t =4.317,P <0.01 ).The level of LC3B-II/I in hippocampus nerve cells of rats in NT and HT groups were significantly increased compared to BC group (F=6.584,P<0.05),while the levels of LC3B-II/I in rats of HT-12 and HT-24 subgroups were significantly reduced compared to NT-12 and NT-24 subgroups,respectively (t=10.836,P<0.001;t=2.653,P=0.02).Ultrastructure damage of nucleus and mitochondria in NT group was more evident compared to BC group,and eumorphism of nucleus and mitochondria were maintained in rats of HT group compared to NT group.Conclusions The mild hypothermia reduced the injury of nerve cells and improved the neurological function of rats survived from cardiac arrest likely by reducing ROS production of nerve cells and inhibition the expression of caspase-3mRNA and lowering the level of LC3 leading to reducing cellular apoptosis and massive autophagy in rats survived from cardiac arrest after CPR.

Objective To study the effect of bromocriptine(BRC) on the activation of T lymphocyte stimulated by phytohemagglutinin(PHA).Methods After CD4+ T cell line Jurkat E6-1 cells were stimulated by PHA,prolactin(PRL) and BRC,respectively,the expression of linker for activation of T cells(LAT) and zeta-chain T cell receptor associated protein kinase 70 000(ZAP-70) mRNA of T lymphocytes were checked by RT-PCR.The expression of PRL mRNA of T lymphocytes was detected by Real time PCR.The expression of CD25(cluster of differentiation) as a marker of early activation on the surface of T lymphocytes was detected by flow cytometry,and the activation of nuclear factor-?B(NF-?B) was detected by luciferase reporter system.Results 1.BRC inhibited the expression of ZAP-70 as the common signal molecules both in the T lymphocyte activation pathway and PRL-prolactin-prolactin receptor(PRLR) signal transduction pathway,and decreased the expression of PRL mRNA produced by activation T lymphocytes.2.BRC enhanced the expression of LAT mRNA as another important signal molecular on the T lymphocytes and CD25 on the surface of the T lymphocytes.3.The activation of NF-?B of T lymphocytes was decreased.Conclusions BRC might inhibit the activation of T lymphocytes by inhibiting the expression of ZAP-70,the common signal molecules between T lymphocytes activation and PRL-PRL pathway,and PRL mRNA,the like-T lymphocyte growth factor.

Objective To study the application of pediculated skin flaps in the treatment of com-plicated long urethratresia. Methods From March 1999 to May 2006, a total of 18 male patients with complicated long urethratresia were treated by using the pediculated skin flaps. The causes of urethratresia were 7 cases of postoperative pelvic fractures with posterior urethral stricture, 4 cases of transurethral intravesical chemotherapy, 3 cases of postoperative bulbar urethral stricture, 2 cases of gonorrhea, and 2 cases of long-time urethral catheter placement. Four cases were urethratresia nf cor-pus penis, 7 cases were anterior urethral obliteration, 7 cases were posterior urethral and anterior ure-thral obliteration. Urethro-perineal fistulas were found in 8 cases, posterior urethrorectal fistulas in 7 cases, false passage formations in 8 cases. The average length of urethratresia was 15.1 cm (range 8. 7 to 23. 0 cm). The urethral scar was rasected, the posterior urethrorectal fistula was repaired, and different kinds of pediculated skin flaps depending on the length of urethratreaia was used. Results All the patients were followed up for 12 to 18 months (mean 14 months). Fifteen patients voided well 3 months postoperatively, none of the urography showed stricture. The mean peak urinary flow rate was 16. 9 ml/s (range from 16. 5 to 21.7 ml/s). Of the other 3 cases, 1 case experienced difficult voi-ding due to the long and circuitous tabularized skin flap but recovered after proper shortening;1 case had restenosis for the infection of anastomosis but voided well after excision and reanastomnsis;1 casehad a urinary fistula resulting from hematoma and infection, but was successfully treated by the neo-plasty of the urinary fistula. The mean peak urinary flow rate was 17.0 ml/s (range 15.0 to 22.0 ml/s) for 17 patients 6 months postoperatively, except for one who experienced genuine urinary incon-tinence. At 9-18 months after operations, the mean peak urinary flow rate was 17.5 ml/s (range 15.8 to 22.5 ml/s) for 17 patients. Conclusion The single-stage urethroplasty based on pediculated skin flaps is a reliable and durable method for complicated long urethratresia.

The purpose of this study was to investigate the efficacy of treatment with haploidentical donor's lymphocyte infusion(hiDLI) combined with interleukin-2 (IL-2) after transplantation of autologous peripheral blood stem cells mixed with haploidentical allogeneic bone marrow (mix-HSCT) for acute myeloid leukemia (AML). 49 patients diagnosed as AML were enrolled in this study. After preconditioning with TBI plus VEMAC regimen, all patients received mix-HSCT. Autologous peripheral blood hematopoietic stem cells were mobilized with chemotherapy-combined G-CSF, and haploidentical allogeneic bone marrow cells were not mobilized with G-CSF. 33 patients in test group were treated with hiDLI plus IL-2 for 1-8 times after hematopoietic reconstruction, 16 patients in control group received mix-HSCT only. All the patients were followed-up for more than 3 years. The results showed that all the patients obtained hematopoietic reconstruction, and no graft-versus-host disease (GVHD) was found. In two groups, the median time of absolute neutrophil count (ANC) ≥ 0.5×10(9)/L was 14 (12 - 18) and 14 (11 - 16) days, and WBC count ≥ 4.0×10(9)/L was 17 (16 - 22) and 18(17 - 20) days, Plt count ≥ 50×10(8)/L were 25 (24 - 29) and 25 (23 - 26) days. 9 patients in test group formed mixed chimerism (46XX/46XY) and sustained about 3 - 12 months; disease-free survival (DFS) was 63.6%, 3 patients in control group formed mixed chimerism (46XX/46XY), persistent about 3-6 months; DFS was 50.0%. It is concluded that treatment with hiDLI plus IL-2 after mix-HSCT for AML patients may increase DFS efficiently.
Adolescent ; Adult ; Female ; Hematopoietic Stem Cell Mobilization ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunotherapy, Adoptive ; Leukemia, Myeloid, Acute ; therapy ; Male ; Transplantation, Homologous ; Young Adult

BACKGROUNDAlveolar echinococcosis (AE) is a disease in human and animals, and the cure rate is unsatisfactory. This study aimed to investigate the curative efficacy of different doses of locally applied radiotherapy on alveolar echinococcosis in rats.

METHODSRats infected with Echinococcus multilocularis were randomly divided into 4 groups of 15 rats each: low-, middle-, and high-irradiation groups and a control group. Rats in the control group underwent no treatment, while rats in the irradiation groups received 6-MeV radiotherapy at 20 Gy/8 f, 40 Gy/8 f, and 60 Gy/8 f respectively, once every 3 days for a total of 8 times. One month after radiotherapy, wet weight and AE vesicle inhibitory rate were detected in rats of each group. Histopathologic and ultrastructural observations of tissues with AE lesions were performed.

RESULTSIn the treatment groups, an obvious inhibitory effect was found in AE rats; the inhibitory rates were 50%, 72%, and 82%, respectively. There were also statistical differences in pathological changes and average wet weight of the lesions compared with the control group (P < 0.05). In the treatment groups, injuries of various degrees were found in the ultrastructure of the laminated and germinal layers in the capsular wall of AE, and injury was most severe in the high-dose group.

CONCLUSIONRadiotherapy has a dose-dependent inhibitory effect on the growth of AE.

Animals ; Dose-Response Relationship, Radiation ; Echinococcosis, Hepatic ; pathology ; radiotherapy ; Female ; Rats

Objective: To investigate the safety, efficacy and prognosis of antegrade dissection re-entry (ADR) with the assistance of BridgePoint devices in opening coronary chronic total occlusion (CTO). Methods: A total of 87 consecutive patients, who underwent percutaneous coronary intervention using BridgePoint devices from April 2016 to December 2018 in Xijing Hospital, were included in this study. General information of the selected patients, features of CTO lesions and intraoperative parameters were recorded. Short-term outcomes including technical success rate (defined as achieving TIMI 3 blood flow with residual stenosis<30%), surgical success rate (defined as no major adverse cardiovascular events (MACE) occured while hospitalized), complications, and MACE during hospitalization were observed. MACE included death, recurrent myocardial infarction, target vascular reconstruction (TVR) and cardiac tamponade. Patients were followed up by outpatient or telephone visits at 30 days and 6, 12, 24 and 36 months after discharge. Results: Eighty-seven patients, aged (61±10) years with J-CTO scores (2.49±0.52) were included, and 75(86%) were male. Six patients underwent direct ADR with BridgePoint system, and all were successful. Eighty-one patients underwent rescue ADR using BridgePoint devices, and 62 of them were successful. The success rate of ADR with BridgePoint devices was 78.2% (68/87). Nine out of the 19 failed cases succeeded after the application of rescue antegrade/retrograde technique. The technical success rate was 88.5% (77/87). Coronary perforation occurred in 2 cases (2.3%), one case was treated with covered stent and the other case with tamponade was treated with pericardiocentesis. One patient developed periprocedural myocardial infarction, and one patient suffered from sudden death, and one patient had cardiac tamponade. In-hospital MACE occurred in 3 (3.4%) patients. The surgical success rate was 85.1% (74/87).The procedure time was (175±72)minutes and the amount of contrast used was (449±155)ml. During a follow-up of 17(11, 26) months, the incidence of MACE within 30 days was 4.7% (4/86), while 10.5% (9/86) within 6 months, 17.4% (15/86) within 17 months. Conclusion: Opening CTO with the assistance of BridgePoint devices is feasible and safe, with high success rate and satisfactory outcome.
Aged ; Chronic Disease ; Coronary Angiography ; Coronary Occlusion ; Humans ; Male ; Middle Aged ; Percutaneous Coronary Intervention ; Risk Factors ; Time Factors ; Treatment Outcome

BACKGROUND: Cardiac arrest (CA) is a common and serious event in emergency medicine. Despite recent improvements in resuscitation techniques, the survival rate of patients with CA is unchanged. The present study was undertaken to observe the effect of mild hypothermia (MH) on the reactive oxygen species (ROS) and the effect of neurological function and related mechanisms. METHODS: Sixty-five healthy male Sprague Dawley (SD) adult rats were randomly (random number) divided into 2 groups: blank control group (n=5) and CPR group (n=60). CA was induced by asphyxia. The surviving rats were randomly (random number) divided into two groups: normothermia CPR group (NT) and hypothermia CPR group (HT). Normothermia of 37 °C was maintained in the NT group after return of spontaneous circulation (ROSC), hypothermal intervention of 32 °C was carried out in the HT group for 4 hours immediately after ROSC. Both the NT and HT groups were then randomly divided into 2 subgroups 12 hours and 24 hours after ROSC (NT-12, NT-24, HT-12, HT-24 subgroups). During observation, the neurological deficit scores (NDSs) was recorded, then the bilateral hippocampi were obtained from rats' head, and monoplast suspension of fresh hippocampus tissue was made immediately to determine the level of intracellular ROS by flow cytometry. Transmission electron microscope was used to observe the ultramicro changes of cellular nucleus and mitochondria. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the expression of caspase-3 mRNA, and western-blotting (WB) was used to determine the level of LC3 in frozen hippocampus tissue. Measured data were analyzed with paired sample t test and One-Way ANOVA. RESULTS: Of 60 rats with CA, 44 (73%) were successfully resuscitated and 33 (55%) survived until the end of the experiment. The NDSs of rats in the NT and HT groups were more significantly reduced than those in the BC group (F=8.107, P<0.05), whereas the NDSs of rats in the HT-12 and HT-24 subgroups were significantly increased in comparison with those NDSs of rats in the NT-12 and NT-24 subgroups, respectively (t=9.692, P<0.001; t=14.374, P<0.001). The ROS in hippocampus nerve cells in the NT and HT groups significantly increased compared to the BC group (F=16.824, P<0.05), whereas the ROS in the HT-12 and HT-24 subgroups significantly reduced compared with that ROS in the NT-12 and NT-24 subgroups, respectively (t=9.836, P<0.001;t=7.499, P<0.001). The expression of caspase-3 mRNA in hippocampus nerve cells in the NT and HT groups were significantly increased compared to the BC group (F=24.527, P<0.05), whereas the expression of caspase-3 mRNA in rats of the HT-12 and HT-24 subgroups was significantly reduced compared to the NT-12 and NT-24 subgroups, respectively (t=6.935, P<0.001; t=4.317, P<0.001). The expression of LC3B-II/I in hippocampus nerve cells of rats in the NT and HT groups significantly increased compared to the BC group (F=6.584, P<0.05), whereas the expression of LC3B-II/I in rats of the HT-12 and HT-24 subgroups significantly reduced compared to the NT-12 and NT-24 subgroups, respectively (t=10.836, P<0.001; t=2.653, P=0.02). Ultrastructure damage of nucleus and mitochondria in the NT group was more evident than in the BC group, and eumorphism of nucleus and mitochondria were maintained in rats of the HT group compared with the NT group. CONCLUSION: Mild hypothermia lessened the injury of nerve cells and improved the neurological function of rats that survived from cardiac arrest by reducing the ROS production of nerve cells and inhibiting the expression of caspase-3 mRNA and LC3, leading to cellular apoptosis and massive autophagy in rats that survived from cardiac arrest after CPR.

BACKGROUNDSpinocerebellar ataxia type 3 (SCA3) is the most common subtype of SCA worldwide, and runs a slowly progressive and unremitting disease course. There is currently no curable treatment available. Growing evidence has suggested that nerve growth factor (NGF) may have therapeutic effects in neurodegenerative diseases, and possibly also in SCA3. The objective of this study was to test the efficacy of NGF in SCA3 patients.

METHODSWe performed an open-label prospective study in genetically confirmed adult (>18 years old) SCA3 patients. NGF was administered by intramuscular injection (18 μg once daily) for 28 days consecutively. All the patients were evaluated at baseline and 2 and 4 weeks after treatment using the Chinese version of the scale for assessment and rating of ataxia (SARA).

RESULTSTwenty-one SCA3 patients (10 men and 11 women, mean age 39.14 ± 7.81 years, mean disease duration 4.14 ± 1.90 years, mean CAG repeats number 77.57 ± 2.27) were enrolled. After 28 days of NGF treatment, the mean total SARA score decreased significantly from a baseline of 8.48 ± 2.40 to 6.30 ± 1.87 (P < 0.001). Subsections SARA scores also showed significant improvements in stance (P = 0.003), speech (P = 0.023), finger chase (P = 0.015), fast alternating hand movements (P = 0.009), and heel-shin slide (P = 0.001).

CONCLUSIONSOur preliminary data suggest that NGF may be effective in treating patients with SCA3.

Adult ; Animals ; Female ; Humans ; Injections, Intramuscular ; Machado-Joseph Disease ; drug therapy ; Male ; Mice ; Middle Aged ; Nerve Growth Factor ; administration & dosage ; therapeutic use ; Prospective Studies

Objective To construct the pEGFP-C1-CXCL1 eukaryotic expression vector and to investigate the effect of CXCL1 on the proliferation of HepG2 cells under endoplasmic reticulum stress ( ERS).Methods Fragments of CXCL1 were obtained from the cDNA library of HepG2 cells before CXCL1 was cloned into a pEGFP-C1 vector for a recombinant plasmid pEGFP-C1-CXCL1 which was screened and identified by PCR and sequence alignment .Then,the recombinant plas-mid of pEGFP-C1-CXCL1 was transfected into human 293 T cell line and the expression of CXCL 1 was detected by fluores-cence microscopy and Western blotting.pEGFP-C1-CXCL1was furhter transfected into HepG2 cells, and CCK8 was used to detect the inhibitory effect of CXCL1 on tumor proliferation induced by TM in hepatocellular carcinoma .Results pEGFP-C1-CXCL1 was vertified by sequencing analysis .Fluorescence microscopy showed that pEGFP-C1-CXCL1 was transfected into 293T.CXCL1 expression was detected by Western blotting .CCK8 showed that TM inhibited tumor proliferation , while overexpression of CXCL1 decreased the inhabitory rate on cell proliferation of HepG 2 cells under ER stress compared to pEGFP-C1 group and the control group .Conclusion A recombinant pEGFP-C1-CXCL1 plasmid is successfully constructed that can be expressed stably in human 293T cells.Overexpression of CXCL1 can effectively reduce the inhabitory rate of HCC cells induced by the ER stress.