Objective To explore the clinical efficacy of laparoscope-assisted transanal total mesorectal excision (La-TaTME) for middle-low rectal cancer.Methods The retrospective cross-sectional study was conducted.The clinical data of 16 patients with middle-low rectal cancer who underwent La-TaTME in the Peking University Third Hospital from August 2015 to August 2016 were collected.Sequential surgery of La-TaTME was applied to patients in the same team,with laparoscopic surgery first and then transanal surgery.Observation indicators:(1) operation and postoperative recovery situations:conversion to open surgery,anastomosis method,operation time,volume of intraoperative blood loss,intraoperative complications,time for out-of-bed activity,time for liquid diet intake,postoperative complications and duration of postoperative hospital stay.(2) postoperative pathological situations:length of surgical specimen,tumor diameter,distance from tumor to resected distant intestinal canal,complete degree of mesorectum,circumferential resection margin,pathological T stage,pathological N stage,number of lymph node detected and tumor cell differentiation.(3) follow-up.Patients in stage Ⅲ-ⅣV of TNM stage of RC underwent postoperative adjuvant chemotherapy.Follow-up using outpatient examination was performed once every 3 months postoperatively to detect the patients' survival and tumor recurrence up to December 2016.Measurement data were represented as M (range).Results (1) Operation and postoperative recovery situations:all the 16 patients underwent successful La-TaTME without conversion to open surgery,including 10 with colorectal anastomosis,3 with colon-canalis analis anastomosis and 3 with permanent colostomy.Operation time and volume of intraoperative blood loss were 290 minutes (range,215-420 minutes) and 50 mL (range,30-100 mL),respectively.One patient had intraoperative complication,showing broken ends ischemia of sigmoid colon after dragging out resected rectum from the anus,following free splenic flexure of colon,about 5 cm ischemic sigmoid colon were resected,and descending colon-rectum anastomosis was performed.Time for out-of-bed activity and time for liquid diet intake were 1 days (range,1-3 days) and 2 days (range,1-9 days),respectively.Among 3 patients with postoperative complications (Ⅱ stage of ClavienDindo),2 with incomplete intestinal obstruction were improved by gastrointestinal decompression and total parenteral nutrition,and 1 with presacral infection was improved by drainage and antibiotic therapy.Duration of postoperative hospital stay was 7 days (range,5-21 days).(2) Postoperative pathological situations:length of surgecal specimen,tumor diameter and distance from tumor to resected distant intestinal canal were respectively 18.0 cm (range,12.0-24.0 cm),3.5 cm (range,0.5-6.8 cm) and 2.5 cm (range,1.0-5.0 cm).Evaluation of mesorectum of surgical specimen:14 patients had complete mesorectum of surgical specimen and 2 had nearly complete mesorectum.There was no residual tumor at circumferential resection margin,proximal and distal ends.Pathological T stage of 16 patients:T0 (pathological complete response after neoadjuvant therapy),T1,T2 and T3 stages were found in 1,1,4 and 10 patients,respectively.Pathological N stage:12,2 and 2 patients were detected in N0,N1 and N2 stages,respectively.Number of lymph node detected was 16 (range,6-32).Tumor cell differentiation:no tumor cell (pathological complete response after neoadjuvant therapy),high-,moderateand low-differentiated tumors were respectively detected in 1,2,7 and 6 patients.(3) Follow-up.All the patients were followed up for 12 months (range,4-16 months).There were no local tumor recurrence or distant metastasis and death.Conclusion La-TaTME may be a new,safe and effective resection for middle-low rectal cancer.

OBJECTIVETo investigate the feasibility of bone marrow mesenchymal stem cell (MSC) transplantation with ultrasound-targeted microbubble destruction.

METHODSTwenty-one Wistar rats were divided into MSCs-iv group (MSCs-iv), ultrasound+MSCs-iv group (US+MSCs-iv), ultrasound+microbubble+MSCs-iv group (US+MB+MSCs-iv) with intravenous MSC transfer, ultrasound and microbubble treatment as indicated. The skeletal muscles were obtained from the rats for microscopic examination with HE staining. The hindlimb gracilis and semimembranosus muscles were sampled 7 days after MSC transplantation, and the transplanted MSCs were detected by immunohistochemistry. The vital organs were collected from rats in US+MB+MSCs-iv group for immunohistochemistry.

RESULTSIn US+MB+MSCs-iv group, HE staining demonstrated the presence of red blood cell leakage into the tissue space in the gracilis and semimembranosus muscles, and immunohistochemistry identified large numbers of transplanted MSCs in the the gracilis and semimembranosus muscles and the spleen, whereas no labeled cells were detected in the skeletal muscles in other groups.

CONCLUSIONUltrasound-targeted microbubble destruction provides a useful means for enhancing the efficiency of stem cell transplantation.

Animals ; Bone Marrow Cells ; cytology ; Cell Movement ; radiation effects ; Female ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; Microbubbles ; Muscle, Skeletal ; cytology ; Rats ; Rats, Wistar ; Ultrasonics

OBJECTIVETo construct a eukaryotic expression vector containing aldehyde dehydrogenase-2 (ALDH2) gene and investigate the effects and its possible mechanisms of ALDH2 gene on cell proliferation and anti-oxidative damage in the K562 cells.

METHODSAn eukaryotic expression vector containing the ALDH2 gene cloned from human hepatocytes was constructed and transfected into K562 cells by liposome. RT-PCR and Western blot were used to evaluate the expression of ALDH2. MTT assay was used to check the cell proliferation and trypan blue exclusion to check K562 cells damage induced by hydrogen peroxide (H2O2). RT-PCR and fluorescence spectrophotometry were used to determine the expression of heme oxygenase-1 (HO-1) and the generation of intracellular reactive oxygen species (ROS) respectively.

RESULTSRT-PCR and Western blot analysis showed distinct higher ALDH2 protein expression in gene transfected group. The latter group had a higher cell proliferation (P < 0.05) and survival rate against H2O2 induced-oxidative damage, being increased by 7.8 times (IC(50) was 12.3 µmol/L and 1.4 µmol/L for K562-pcDNA3.1-ALDH2 and control cells, respectively, P < 0.01). The HO-1 mRNA expression and the generation of intracellular ROS were downregulated at a specific concentration of H2O2 in the ALDH2 gene transfected group.

CONCLUSIONALDH2 gene transfection can protect K562 cells against oxidative damage, and the downregulation of HO-1 expression and intracellular ROS may be involved in this process.

Aldehyde Dehydrogenase ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Hydrogen Peroxide ; K562 Cells ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo suppress the expression of CCR5 and CXCR4, the co-receptors for human immunodeficiency virus type 1 ( HIV-1), and thus inhibit HIV-1 from entering cells.

METHODSDNA fragments encoding either CCR5 or CXCR4 were amplified from healthy human peripheral blood mononuclear cells (PBMCs) by reverse transcript polymerase chain reaction (RT-PCR) and sequencing was performed. Correct fragments were inserted into Shuttle plasmid inversely, which was recombined with backbone plasmid containing homologous adenoviral genome in E. coli BJ5183. The recombinant plasmids were transfected into 293 cells in which they were packaged and amplified. Recombinant adenoviruses containing antisense RNA of CCR5 or CXCR4 were obtained and identified by RT-PCR, and the titres of them were determined by cytopathic effect (CPE) method. The U937 and MT4 cells were infected by recombinant adenoviruses containing antisense RNA of CCR5 (multiplicity of infection, MOI = 100) and CXCR4 (MOI = 200), respectively. The expression of co-receptors on infected cell was measured by fluorescence activated cell sorter at 24, 48, 72 hours and 10 days after infection. In addition, the chemotactic activity and proliferation of infected cells were detected with Boyden chamber and 3H incorporation respectively.

RESULTSWe constructed the recombinant plasmids and obtained the recombinant adenoviruses which contained antisense RNA of CCR5 or CXCR4 and were designated as pAd-antiR5 and pAd-antiX4 respectively. The titers of recombinant adenoviruses pAd-antiR5 and pAd-antiX4 were 5 x 10" PFU/ml and 7 x 10(10) PFU/ml, respectively. The expression rate of CCR5 on U937 cells decreased from 82. 10% (blank control) to 1.12% (Ad-antiR5 infected) , and that of CXCR4 on MT4 cells decreased from 42% (blank control) to 1.03% (Ad-antiX4 infected) 24 hours later. The expression rates of CCR5 on Ad-antiR5 infected U937 cells were 1.02% , 1.26% , 1.23% at 48 hours, 72 hours, and 10 days later, respectively. The expression rates of CXCR4 on Ad-antiX4 infected MT4 cells were 1.13%, 1.17%, 1.22% at 48 hours, 72 hours, and 10 days later, respectively. Moreover, the recombinant adenovirus had no effects on chemotactic activity and proliferation of the cells.

CONCLUSIONThe recombinant adenovirus containing antisense CCR5 or CXCR4 can remarkably decrease the expression of co-receptors for HIV-1 on U937 or MT4 cells without affecting their chemotactic activities and proliferative abilities.

Adenoviridae ; genetics ; Cell Line, Transformed ; Cell Proliferation ; Chemotaxis ; Down-Regulation ; Genetic Vectors ; Humans ; RNA, Antisense ; genetics ; RNA, Messenger ; genetics ; Receptors, CCR5 ; biosynthesis ; genetics ; Receptors, CXCR4 ; biosynthesis ; genetics ; Transfection ; U937 Cells

OBJECTIVESTo evaluate the relationship between the functional integrity of sperm membrane and seminal parameters related to CASA.

METHODSThirty-eight fertile and one hundrend and twenty four infertile males were tested the functional integrity of sperm membrane by the kit and parameters by CASA.

RESULTSThere was a significant difference in the functional integrity of sperm membrane between fertile and infertile group (P < 0.01). The items related to CASA between normal and abnormal group in the functional integrity of sperm membrane had a remarkable difference, except motion degree, seminal volume and pH.

CONCLUSIONSTo determine the functional integrity of sperm membrane can be used as a necessary supplementary method for CASA, and it has clinical significance in diagnosing, treating and researching male infertility.

Adult ; Humans ; Image Processing, Computer-Assisted ; Male ; Membranes ; physiology ; Semen ; physiology ; Spermatozoa ; physiology

Objective To evaluate the effect of schistosomiasis control and prevention in Yangxin County from 2004 to 2015,so as to provide the evidence for improving the work of schistosomiasis transmission interrupted and elimination in the fu-ture. Methods According to the endemic types and endemic regularity of schistosomiasis in Yangxin County,the comprehen-sive control strategies were adopted,and the programs related to sanitation,water conservancy,forestry,and agriculture were implemented continuously. The schistosomiasis control effects in this county from 2004 to 2015 were analyzed and compared. Re-sults After the implementation of the schistosomiasis comprehensive control strategies in Yangxin County ,the calculated num-ber of patients reduced from 22240 in 2004 to 1471 in 2015,the infection rate of residents reduced from 8.57%in 2004 to 0.16%in 2015,the number of patients with acute schistosome infection reduced from 64 in 2004 to 0,and no cases of acute schistosomiasis found since 2009. The infection rate of cattle decreased from 8.87%in 2004 to 0. The area with Oncomelania hu-pensis snails and the area of susceptible zone reduced from 3446.21 hm2 and 1111.59 hm2 in 2004 to 2285.75 hm2 and 41.28 hm2 in 2015 respectively,and the schistosome-infection rate of snails reduced from 0.76%in 2004 to 0. Conclusions Since the comprehensive control strategy implemented from 2004,the endemic situation of schistosomiasis in Yangxin County has de-creased significantly. However,the harness force of the Fu River as well as the control of infection source of livestock still should be strengthened to consolidate the control achievement.

BACKGROUNDThe mechanism of acupuncture analgesia in craniotomy has been widely studied. However, the theoretical basis for selection of acupoints has not been examined. In this study, we used the regional homogeneity method blood oxygen level-dependent (BOLD) signals to determine changes in brain activity in response to transcutaneous electrical stimulation on acupoints and non-acupoints in resting state functional magnetic resonance imaging (fMRI).

METHODSTwelve healthy volunteers were enrolled in this study. BOLD fMRI scanning of the brain was performed for 306 seconds before and 30 minutes after transcutaneous electrical stimulation on acupoints UB63 (Jinmen), LV3 (Tai chong), ST36 (Zusanli), and GB40 (Qiuxu). The procedure was repeated after one week with stimulation on non-acupoints (one was 9 above BL67, the second was 12 above BL67 (Kunlun), the third was 7 above KI3, and the fourth was 10 above KI3 (Taixi)).

RESULTSThe regional homogeneity in the acupoint group was increased in the left thalamus, caudate, putamen, lentiform nucleus (BA19, 30, 39), postcentral gyrus, precentral gyrus (BA3, 4, 30, 32), calcarine fissure, middle temporal gyrus (BA30), right superior temporal gyrus, inferior temporal gyrus (BA38), cuneus, and precuneus (BA7, 19) when compared to the non-acupoint group. The regional homogeneity of the acupoint group was decreased in the left cerebellum posterior lobe, middle frontal gyrus (BA10), double-side precuneus (BA7), and the postcentral gyrus (BA40).

CONCLUSIONSThe brain region activated following acupoint stimulation is the ipsilateral pain-related brain region, which may relate to the therapeutic effect of acupuncture on pain relief. Further acupoint stimulation causes different central nervous responses compared to non-acupoint stimulation.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Transcutaneous Electric Nerve Stimulation ; Young Adult

Objective To understand the knowledge and attitudes towards second hand smoking(SHS) among restaurant or bar patronage in five cities in China. Methods 405 restaurants and bars were conveniently selected in 5 cities as Beijing, Wuhan, Xi'an, Kunming and Guiyang. From each of these hospitality venues, at least 3 patrons, including one smoker, one female non-smoker and one male nonsmoker were asked to answer a questionnaire on their knowledge and attitudes towards SHS and smoking policies in public places. Results 43.1% of the respondents had good knowledge on SHS hazards. 65%reported that those who smoked around them had never asked for their permission, and about 60%admitted that they had never asked others to stop smoking in front of them. There were almost two thirds of patrons reporting that they once felt discomfort of SHS in restaurants or bars, nearly half of whom chose free restaurants and bars' initiations were only 30.0% and 19.8%, respectively. In Beijing, logistic regression model analyses showed that those who supporting 100 % smoke-free restaurants and bars tended to be non-smokers, aged 25 years or older and to have had at least college education. Also in Beijing, those being female, having at least college education or non-smokers were more likely to choose restaurants and bars with smoking restrictions. Conclusion Though knowledge on SHS hospitality patronage was still not high or popular for the public to refuse SHS exposure, banning smoking in hospitality venues was public's inclination and would become a trend in public health endeavor.

Animals ; Cellular Reprogramming ; Dentate Gyrus ; cytology ; Fibroblasts ; cytology ; Mice ; Neural Stem Cells ; cytology ; transplantation ; Swine

This study aims to explore the mechanism of fresh Phragmitis Rhizoma against chronic bronchitis airway inflammation. The SD rats of SPF grade were divided into control group, model group, Guilongkechuanning group(GLKCN, 1.125 g·kg~(-1)), high-dose fresh Phragmitis Rhizoma group(LG-HD, 15 g·kg~(-1)), and low-dose fresh Phragmitis Rhizoma group(LG-LD, 7.5 g·kg~(-1)). The chronic bronchitis models of rats in other groups except the control group were induced by the modified smoking method. From the 15 th day of modeling, the rats were given corresponding agents by gavage for 20 consecutive days. After the last administration, the rats were sacrificed for sample collection. Enzyme-linked immunosorbent assay(ELISA) was employed to detect serum transforming growth factor-β(TGF-β) and interleukin-6(IL-6) levels. The protein expression of TGF-β, IL-1β and IL-6 in lung tissue was detected by immunohistochemical method. Masson staining was performed to detect collagen fibers and muscle fibers in lung tissue, and HE staining to detect the pathological changes of lung tissue. Human bronchial epithelial(16 HBE) cells were cultured in vitro, and CCK-8(cell counting kit-8) method was used to detect the cytotoxicity of cigarette smoke extract(CSE) and fresh Phragmitis Rhizoma. After the exposure of 16 HBE cells to 3.5% CSE and appropriate concentration(800, 400 μg·mL~(-1)) of fresh Phragmitis Rhizoma for 24 h, quantitative real-time PCR was conducted to determine the mRNA levels of TGF-β and IL-1β, and Western blot was employed to determine the protein levels of TGF-β and IL-6 in the cells. The rat model of chronic bronchitis induced by smoking was successfully established. Fresh Phragmitis Rhizoma reduced serum TGF-β and IL-6 levels, down-regulated the protein levels of TGF-β, IL-1β, and IL-6 in lung tissue, and alleviated pathological changes and fibrotic lesions in lung tissue. Moreover, it down-regulated the CSE-induced protein expression of TGF-β and IL-6 as well as the mRNA level of TGF-β in 16 HBE cells. These results indicated that fresh Phragmitis Rhizoma could prevent airway inflammation from chronic bronchitis and promote cell repair by inhibiting the TGF-β signaling pathway.
Animals ; Bronchitis, Chronic/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Inflammation ; Lung ; Poaceae/chemistry* ; Rats ; Rats, Sprague-Dawley ; Rhizome ; Signal Transduction ; Transforming Growth Factor beta/genetics*