Objective To establish a simple and rapid method for the extraction and detection of residual acetochlor and atrazine in environment. Methods Acetochlor and atrazine in environment were extracted and enriched efficiently by using automatic solid phase extraction system. The sample concentrations were determined by gas chromatography-mass spectroscopy with electron impact ionization and multipolar mass spectra Msn. Results The recovery rates and RSDs of the method were 79.2%-95.1% and 1.73%-8.31% respectively. The detection limit of acetochlor in water and soil were 0.1 ?g/L and 0.005 mg/kg respectively. The detection limit of atrazine in water and soil were 0.05 ?g/L and 0.002 5 mg/kg respectively. Linear range was 0-5 ?g/ml. Conclusion The SPE-GC-MS method can be used to determine acetochlor and atrazine in environment.

Objective To evaluate the role of autophagy in the development of inflammatory pain in the rats.Methods Twenty-four healthy adult male Sprague-Dawley rats,weighing 180-240 g,were randomly divided into 4 groups (n=6 each) by using a random number table:control group (group C),inflammatory pain group (group IP),dimethyl sulfoxide (DMSO) group (group D),and rapamycin (autophagy inducer) group (group R).Inflammatory pain was produced by injecting 50 μl bee venom into the plantar surface of the left hindpaw.In group C,0.9％ normal saline was injected into the plantar surface of the left hindpaw.In group D,2％ DMSO was injected through a gastric tube into the stomach 1 ml per day for 3 consecutive days,and the model was established at 1 h after injection on 3rd day.In group R,rapamycin l0 mg/kg (in 2％ DMSO) was injected through a gastric tube into stomach 1 ml per day for 3 consecutive days,and the model was established at 1 h after injection on 3rd day.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 2 h after the model was established.After measurement of the pain threshold,the dorsal horn of the spinal cord was removed for determination of the expression of microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ),Beclin-1 and p62 by Western blot.Results Compared with group C,the MWT was significantly decreased,the TWL was significantly shortened in IP and D groups,and the expression of LC3 Ⅱ,Beclin-1 and p62 in the dorsal horn of the spinal cord was significantly up-regulated in IP,D and R groups (P＜0.05 or 0.01).Compared with group IP,the MWT was significantly increased,the TWL was significantly prolonged,the expression of LC3 Ⅱ and Beclin-1 in the dorsal horn of the spinal cord was significantly up-regulated,and the expression of p62 was significantly down-regulated in group R (P＜0.05),and no significant change was found in the parameters mentioned above in group D (P＞0.05).Conclusion Autophagy disorders are involved in the development of inflammatory pain in the rats.

Background The pathogenesis of diabetic retinopathy (DR) might be related with oxidative stress and its mechanism has not been fully elucidated.Grape seed proanthocyanidin extracts (GSPE),a powerful antioxidant,plays roles in some systemic diseases.However,the effect and mechanism of GSPE in DR has not been illuminated clearly.Objective This study was to investigate whether GSPE has a protective effect on the retinas of diabetic subjects and explore its mechanism.Methods Thirty SPF adult Wistar rats were divided into the control group,diabetic group and diabetes + GSPE group according to the randomized number table.Diabetic models were induced by intraperitoneal injection of 100 mg/kg strebtozotocin (STZ) prepared with citrate buffer,and only the equal amount of citrate buffer was used in the same way in the control group.GSPE solution was intragastrally used 250 mg/kg daily in the rats of the diabetes+GSPE group from injective day through 8 weeks,and distilled water was used in the same way in the rats of the control and diabetic groups.The rats were sacrificed in the eighth week after injection and the retinas were isolated.The morphology of the retina were examined by hematoxylin and eosin stain.Retinal homogenatewas prepared for the assay of superoxide dismutase (SOD),glutathione peroxidase (GSH-Px) activity and malonaldehyde (MDA) content.The expressions and location of nuclear erythroid related factor 2 (Nrf2) was determined by immunochemistry,and the expressions of Nrf2 and heme oxygenase-1 (HO-1) were quantitatively analyzed by Western blot.The apoptosis of retinal cells was detected by TUNEL.Results In the eighth week after modeling,the blood glucose levels were significantly higher and the body weight was lower in the rats of the diabetic group and diabetes+GSPE group compared with the control group (all at P＜0.01).Retinal structure was normal in the rats of the control group.However,loose tissue,irregular arrangement of cells and thickness decrease of retinal fiber layer,retinal ganglion cell layer and inner plexiform layer were exhibited in the rats of the diabetic group,while the morphology abnormality was slight in the rats of the diabetes+GSPE group.The SOD and GSH-Px activities and MDA content were significantly different among the 3 groups (F =11.010,P =0.001 ; F =12.072,P =0.000 ; F =25.224,P=0.000),and activities of SOD and GSH-Px in the retinas were significantly lower,and the MDA level was higher in the rats of diabetic group than those of the diabetes+GSPE group and the control group (all at P＜0.01).The relative expressing levels of Nrf2 were (165.5±29.4) ％ and (134.8 ±7.8) ％ in the diabetes+ GSPE group and diabetic group,with a significant difference between them (t=2.450,P=0.044).Compared with the diabetic group,the expressing level of the HO-1 was sigficantly increased ([170.2±22.0)％ versus [125.3±9.2] ％,t =2.360,P =0.002).TUNEL showed that the retinal apoptotic cells of diabetic rats were mainly located in the retinal fiber layer,RGCs layer,inner and outer nuclear layer,and the number of apoptotic cells was less in the diabetes+GSPE group compared with the diabetic group under the fluorescence microscope.Conclusions GSPE can play a protective role on diabetic rats by activating Nrf2 pathway and therefore improving retinal oxidative stress and decreasing apoptosis.

Objective To investigate the affection of negative life and events coping style to youthful breast cancer patients and their psychological stress state preoperation. Methods 60 youthful breast cancer patients preoperation were investigated with Life Events Scale (LES) and somatization, depression and anxiety in Self-reporting Inventory Symptom Checklist 90 (SCL-90). Results The score of somatization, depression and anxiety were (1.89 ±0.31), (2.28 ± 0.56) and (1.99 ± 0.63). the anxiety and depression were obvious in patients who had lower education level (P < 0. 05). Sematization, depression and anxiety were more severe in patients who was single, divorced and widowed than married (P < 0. 05). The levels of depression and anxiety were higher in patients who had higher income (P < 0. 05). Conclusions The youthful breast cancer patients have severe psychic trauma, and they need humanistic care.

Objective To evaluate the efficacy of transforanminal endoscopic spine system (TESSYS) technique in treating lumbar disc herniation complicated with Ⅰ degree stability of lumbar spondylolisthesis.Methods Thirty-two patients with lumbar disc herniation,aged 51-82 yr,weighing 52-93 kg,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,were randomly divided into 2 groups (n=16 each) according to whether patients had lumbar spondylolisthesis:lumbar disc herniation group (Y group) and lumbar disc herniation combined with Ⅰ degree stability of lumbar spondylolisthesis group (Y+Z group).Extirpated protrusion,plasty ligamenum flavum and posterior longitudinal ligament and nerve root decompression were carried out using TESSYS technique in two groups,and in addition excision of osseous neoplasias and retro-positioned posterior margin of lumbar vertebral body was done in group Y+Z.Pain was assessed using Visual Analogue Scale (VAS) score at 1 day before surgery and 3 days and 1,3,6 and 12 months after surgery.Patient's function was assessed by using the Oswestry Disability Index (ODI) at 1 day before surgery and 12 months after surgery.The therapeutic effect was evaluated using modified Macnab criteria at 12 months after surgery.Results Compared with the baseline at 1 day before surgery,VAS scores were significantly decreased at each time point after surgery,and ODI was decreased at 12 months after surgery in two groups (P＜0.05).Compared with group Y,VAS scores were significantly decreased at 3 and 6 months after surgery (P＜0.05),and no significant change was found in ODI at each time point or VAS scores and Macnab outcome grade at 12 months after surgery in group Y+Z (P＞ 0.05).Conclusion TESSYS technique can be used to treat lumbar disc herniation complicated with Ⅰ degree stability of lumbar spondylolisthesis.

Purpose To induce the differentiation of hu-man umbilical cord mesenchymal stem cells ( HUCMSCs) into annulus fibrosus (AF) cells by in vitro co-culture technique and to investigate the morphological and histological changes of HUCMSCs after co-culture. Methods HUCMSCs and AF cells were isolated from the normal neonatal umbilical cord and New Zealand white rabbit. Transwell six-well plates were used for co-culture with the cells seeded at the ratio of 1 ∶ 1, HUCMSCs cultured alone served as controls. After two weeks of co-culture, morphological changes were observed by inverted microscope. Real-time PCR was used to detect the expression of typeⅠcolla-gen, aggrecan and SOX-9 gene in HUCMSCs. Immunocyto-chemical staining and toluidine blue staining were used to detect the synthesis of cell matrix such as type Ⅰ collagen and aggre- can. Results The morphology of HUCMSCs in control group was long-fusiform, the morphology of HUCMSCs in co-culture gradually became short-fusiform or polygonal, and began to ap-pear synapse, showing the morphological features of AF-like cells. Real-time PCR results showed that typeⅠcollagen, aggre-can and SOX-9 mRNA were significantly increased in the co-cul-ture group (P＜0. 05). Immunocytochemical staining and tolui-dine blue staining showed that type I collagen and aggrecan were positive, respectively. Conclusion In vitro co-culture technol-ogy can induce HUCMSCs to differentiate into AF-like cells, which is expected to provide a new kind of seed cells for the bio-logical treatment of degenerative disc disease.

OBJECTIVETo explore the efficacy of unrelated umbilical cord blood transplantation (UCBT) in the treatment of Gaucher disease.

METHODThe clinical characteristics of three children with Gaucher disease underwent UCBT in our hospital between April 2013 and September 2014 were retrospectively analyzed. Literature on allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the treatment of Gaucher disease was searched at Wanfang and Pubmed databases between 1983 and 2015 and was reviewed and summaried.

RESULTThree children with Gaucher disease, all were female, received UCBT. These patients' age at receiving transplantation was 3.8 years, 7.1 years and 2.6 years, respectively. The second case received the second transplantation. The first and third case received splenectomy before UCBT. The pretreatment regimen was busulfan (Bu)/fludarabine (Flu)/cyclophosphamide (CTX)/antithymocyte globulin (ATG), and for the patient received the second transplantation melphalan was added to the myeloablative conditioning regimen of Bu/Flu/CTX/ATG. Cyclosporine and mycophenolate mofetil (MMF) wee used for prophylaxis of acute graft versus host disease (aGVHD). The dose of cord blood stem cell nucleated cell counts was 9.7 × 10⁷ /kg,11.9 × 10⁷ /kg and 7.6 × 10⁷/kg respectively. The dose of cord blood stem cell CD34⁺ cell counts was 5.4 × 10⁵/kg , 3.5 × 10⁵/kg and 3.2 × 10⁵/kg respectively. The day of granulocytes exceeding 0.5 × 10⁹/L was day 11, 12 and 19 after transplantation, respectively. The day of platelets exceeding 20 × 10⁹/L was day 14, 33 and 74 after transplantation, respectively. At one month after transplantation the rate of chimerism was over 95% and all patients got donor complete chimerism. The level of β-glucocerebrosidase recovered to normal at one month after transplantation. During transplantation, all patients developed cytomegalovirus (CMV) and Epstein-Barr virus (EBV) viremia. In case 1 immune thrombocytopenia occurred at five month after transplantation unresponding to steroids and mesenchymal stem cells infusion was administered and his platelet in routine blood test recovered to normal. But the patient died because she was infected with varicella-zoster virus out of hospital at nine month after transplantation and the level of β-glucocerebrosidase was normal before death and chronic GVHD (cGVHD) was not found. The case 2 is now in 19th month after transplantation and his level of β-glucocerebrosidase was normal. cGVHD was not found. The patient is currently free of disease. The case 3 was in 9th month after transplantation and his level of β-glucocerebrosidase was normal. cGVHD was found at 112 day after transplantation and was localized and could be controlled by hormonal therapy. The patient is currently free of disease. Three patients' size of liver was significantly reduced after their level of β-glucocerebrosidase ecovered. There were 50 cases with Gaucher disease who were treated with allo-HSCT in the literature and none of them were reported from China. Disease-free survival rate of patients treated with allo-HSCT for Gaucher disease was 85%. In all reports, there were 31 cases who had information of typing of Gaucher disease, of whom 22 cases had type 1 and 9 cases had type 3. Twenty-nine cases had information of survival, of whom 24 cases survived and 5 cases died of infection. Fifteen cases had data of engraftment, 2 of whom had graft failure and one had late graft failure.Glucocerebrosidase recovered to normal in 25 of 31 cases who had relevant data, in one of whom with late graft failure the enzyme recovered to normal 3 month after transplantation, but his enzyme decreased to the initial level 9 month after transplantation. Along with enzyme level's recovery to normal, in a part of cases bone pain and hepatomegaly were relieved and growth delay was improved.

CONCLUSIONThe unrelated UCBT may be a form of treatment that offers the potential of permanent cure and a procedure with possible long-term benefits in patients with Gaucher disease.

Antilymphocyte Serum ; therapeutic use ; Busulfan ; therapeutic use ; Child ; Child, Preschool ; China ; Cyclophosphamide ; therapeutic use ; Cyclosporine ; therapeutic use ; Disease-Free Survival ; Female ; Gaucher Disease ; therapy ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Mycophenolic Acid ; analogs & derivatives ; therapeutic use ; Retrospective Studies ; Transplantation Conditioning ; Vidarabine ; analogs & derivatives ; therapeutic use

Objective To study the epidemiological characteristics of chronic pain following breast cancer surgery and analyze the related factors.Methods Nine hundred and twelve patients following breast cancer surgery were enrolled in the Department of Breast Surgery,Affiliated Hospital,Academy of Military Medical Sciences.The Douleur Neuropathique-4 (DN-4) questionnaire was used to identify neuropathic pain,and the related factors were statistically analyzed.Results The follow-up was completed in 821 patients,including 263 (32％) patients with chronic pain.DN-4 score of 47 (17.9％)patients was over 4 points.There was significant difference between the painful group and the non-painful group in methods of surgery and axillary lymph node dissection (P ＜ 0.05).Conclusion Findings suggest that chronic pain after breast cancer surgery is a significant problem clinically.Proper surgery and psychological buildup are of clinical value in preventing post-surgery chronic pain.

Objective Bladder cancer is one of the most common malignant tumors involving urinary system , yet its pathogene-sis has not been fully and thoroughly studied .The study aimed to de-tect the expression of LASS 2 in bladder cancer model of nude mice and investigate the relationship of LASS 2 with tumor proliferation and apoptosis as well as its possible molecular mechanism . Methods Tumor development in nude mice was observed through the establish-ment of orthotopic bladder cancer model by transplantation , bladder cancer metastasis model by subcutaneous injection and blank con-trol group.LASS2 expression and changes in proliferation and apoptosis were detected in tumor tissues of different parts . Results Bladder cancer cell injected subcutaneously metastasis model tumor formation rate of 100%.The two models were not found transfer phenomenon in vivo.Compared with blank control group (81.0%), LASS2 expression (60.0%, 14.0%) was significantly decreased in the inoculated group and subcutaneous implantation group ( P<0.05) .Compared with the blank control group ( 16.0%) , the expression of Ki67 in the inoculated group and subcutaneous implantation group increased (50.0%and 78.0%) (P<0.05).Compared with the in situ perfusion group, the expression of LASS2 (14.0%) was significantly decreased (P<0.05) and the expression of Ki67 (78.0%) was increased (P<0.05).Compared with the blank control group , the expression of Bcl-2 in subcutaneous implantation group and in si-tu perfusion group was significantly increased ( P<0.05) .Compared with the subcutaneous implantation group , the expression of Bcl-2 was increased in the in situ perfusion group ( P<0.05) , while the expression of Bcl-x1 in the in situ implanted tumor was higher than that in the other two groups (P<0.05).The expression level of Bax and caspase3 in each group was not statistically significant (P>0.05) .Compared with the blank control group , the expression of Bim was significantly decreased in the subcutaneous implantation group (P<0.05). Conclusion The expression of LASS2 may be related to the tumorigenicity , proliferation and apoptosis in EJ blad-der cancer cells .

Transcription factor EB(TFEB)is a member of the MiTF/TFE family and plays an important role in cell stress,metabolism,cancer and so on.There are relatively few studies on the role of TFEB in renal diseases.TFEB was initially found to be highly expressed in TFEB-fusion renal cell carcinoma and plays a key role in the development of re-nal cell carcinoma.Blocking the downstream signaling pathway activated by TFEB would be a promising treatment for TFEB-fusion renal cell carcinoma.On the contrary,the expression of TFEB in renal intrinsic cells is decreased in diabetic kidney disease,leading to a blockage in the autophagy-lysosome pathway.TFEB enhances the ability of cell stress and self-repair,and then delays the progress of diabetic kidney disease.In cystine nephropathy,TFEB expression is reduced in re-nal tubular epithelial cells and compensatory activation is insufficient as well.TFEB over-expression effectively eliminates intracellular cystine and repairs damaged lysosome,which is expected to alleviate or cure the Fanconi syndrome.In summa-ry,TFEB plays a key role in different kidney diseases,and targeted regulation of TFEB provides new hope for the treatment of kidney diseases.