1.Optimal condition of directional-differentiation of neurons from retinoic-acid induced MSESPU35 embryonic stem cell lines
Maolin QIN ; Wenqin CAI ; Zhongxiang YAO
Chinese Journal of Tissue Engineering Research 2006;10(1):157-160
BACKGROUND: Neural axon regeneration is one of the difficulties that must be overcome in treatment of injury of central nerve system. Significant therapeutic effects have been obtained in transplantation of neural stem cells (NSCs), embryonic stem cells (ESCs) and Schwann cells. But the bottleneck situation of insufficiency of cell provider has limited the development on it.OBJECTIVE: To observe directional-differentiation of retinoic-acid induced ESCs so as to find optimal condition for neuronal differentiation.DESIGN: Non-randomized controlled experiment was designed.SETTING: Teaching-Research Room of Histology and Embryology, Department of Basic Medicines. Third Military Medical University of Chinese PLAMATERIALS: The experiment was performed in Staff Room of Histology and Embryology, Third Military Medical University of Chinese PLA from January to May 2000. Eighteen Kunming mice in disoestrus were employed, of which. 12 mice were female and 6 mice male. They were placed in same cage at ratio of 2:1 for mating. The date of pregnancy was recorded. MESPU35 ESC line was prepared.METHODS: Removed head. internal organs and four limbs, feeder-layer Feeder-layer adherent culture was used to proliferate MESPU35 ESCs.Classic 4-/4+ method [The embryoid body (EB) grew naturally for 4 days,without retinoic acid added. In the coming 4 days, retinoic acid was added to induce neural EB of high proportion] was applied to induce the directional differentiation of the nerve. EB was cultured with serum of different concentrations. Phase contrast microscope was used to observe nerve-like EB in serum of different concentrations and to count numbers. ②Immunocytochemical technique was used to observe cellular morphological charac ters at various differentiating phase spots (5th. 9th, 14th days) and with retinoic acid at various concentrations. Flow cytometer (FCM) was used to count the proportion of differentiated neurons.MAIN OUTCOME MEASURES: ①Estimated measurement of the length of process and cell body during formation of neural EB after retinoic-acid induced differentiation of MESPU35 ESCs. ② Observation of cell morphology with immunocytochemical staining and proportion of differentiated cells assayed with FCM.RESULTS: ①It was discovered with phase contrast microscope that serum of different concentrations affect neural directional differentiation after EB formation to certain extent. Excessively high and low concentrations of serum reduced the proportion of neural differentiation of EB. The differentiating proportion is high in serum with 5% concentration. ② It is observed with immunocytochemical technique that the proportions of NF200 positive cell and glial fibrillary acidicprotein (GFAP) positive cell in differentiation of MESPU35 ESCs induced by retinoic acid were increased with phase spots in differentiation and increased concentration of retinoic acid. NF200 positive cell is transformed as multipolar neurons from absence of process in morphology. The processes of GFAP positive cell became longer and linked among each other as reticular pattern finally. ③ It was assayed with FCM that the proportion changes of GFAP positive cell and NF200 positive cell manufactured in differentiation were similar to immunocytochemical one.CONCLUSION: Retinoic acid in combination with proper concentration of serum and differentiating phase spots can induce neural-differentiation of MESPU35 ESC at high proportion and its differentiating regulation is in the patterns of concentration dependence and time dependence.
2.Correlation between mRNA and protein expression of quinine nicotinamide adenine dinucleotide dehydrogenase 1 and heme oxygenase 1 and liver injury due to coal arsenic poisoning
Qi WANG ; Aihua ZHANG ; Maolin YAO ; Peng LUO
Chinese Journal of Endemiology 2017;36(1):21-25
Objective To detect the mRNA and protein expression of downstream quinine nicotinamide adenine dinucleotide (NADH) dehydrogenase 1 (NQO1) and heme oxygenase 1 (HO-1) induced by blood nuclearrelated factor E2 (Nrf2) in the peripheral blood of those exposed to arsenic in the endemic area of coal arsenic poisoning in Guizhou Province,and to discuss its role in the process of occurrence and development of liver injury due to coal arsenic poisoning.Methods Jiaole and Changqing villages in coal-burning-borne arsenism areas in Xingren County of Guizhou were selected as the survey sites,and 161 cases of arsenic-exposed residents were selected as the arsenic exposed group based on physical examination.They were divided into non-patient group (21 cases) and patient group (140 cases) according to the Diagnostic Criteria of Endemic Arsenism (WS/T 211-2001),and the patient group was further divided into mild hepatosis group (52 cases),moderately severe hepatosis group (36 cases) and non-apparent hepatosis group (52 cases) according to the Diagnostic Criteria of Occupational Chronic (GBZ 59-2010).Moreover,45 residents from one village neighboring to non-epidemic area were selected as controls.Peripheral blood samples were collected from all subjects.And mRNA expression of NQO1 and HO-1 were detected by RT-qPCR.Content of NQQ1 and HO-1 were detected by enzyme linked immunosorbent assay (ELISA).Results (①)Results of mRNA expression of NQ01 and HO-1:the relative expression level of mRNA of NQO1 and HO-1 in peripheral blood went up gradually as the degree of liver injury of population exposed to arsenic increased (F =5.548,10.961,all P < 0.05);thereinto,the relative expression level of mRNA of NQO1 of mild hepatosis group (median:0.918) was higher than that of the control group (0.576,P < 0.05),the relative expression level of mRNA of NQO1 of moderately severe hepatosis group (1.243) was higher than those of control group,non-patient group (0.653),non-apparent hepatosis group (0.636) and mild hepatosis group (all P < 0.05);the relative expression level of mRNA of HO-1 of non-patient group (1.059),non-apparent hepatosis group (1.225),mild hepatosis group (1.553) and moderately severe hepatosis group (1.604) were higher than that of control group (0.767,all P < 0.05);the relative expression level of mRNA of HO-1 of mild hepatosis group was higher than that of non-patient group (P < 0.05);the relative expression level of mRNA of HO-1 of moderately severe hepatosis group was higher than those of non-patient group and non-apparent hepatosis group (all P < 0.05).②)Results of protein expression of NQO-1 and HO-1:the level of protein expression of NQO1 and HO-1 in serum went up gradually as the degree of liver injury of population exposed to arsenic increased (F =19.685,17.725,all P < 0.05).Thereinto,the protein expression of NQO1 and HO-1 of non-apparent hepatosis group,mild hepatosis group and moderately severe hepatosis group [NQO1:(6.272 ± 0.744),(6.336 ± 0.628),(6.714 ± 0.540) U/L;HO-1:(45.150 ± 4.813),(45.283 ± 5.049),(48.610 ± 5.365) U/L] were higher than those of control group and non-patient group [NQO1:(5.550 ± 0.730),(5.750 ± 0.427) U/L;HO-1:(39.856 ± 5.249),(42.375 ± 3.014) U/L,all P < 0.05],the protein expression of NQO1 and HO-1 of moderately severe hepatosis group were higher than those of non-apparent hepatosis group and mild hepatosis group (all P < 0.05).Conclusion The expression of mRNA and protein of NQO1 and HO-1 is closely related to the occurrence and development of liver injury due to arsenic exposure in coal arsenic poisoning areas.
3.Expression of thioredoxin reductase 1 in liver and peripheral blood of human and rats exposed to airborne arsenic through coal-burning
Yong HU ; Aihua ZHANG ; Maolin YAO ; Xudong TANG ; Xiaoxin HUANG
Chinese Journal of Endemiology 2015;34(1):12-16
Objective To study the expression and enzyme activity of thioredoxin reductase 1 (TrxR1) in liver and peripheral blood of human and rats exposed to airborne arsenic through coal-burning as well as its role in liver injury of coal-burning-borne arsenic poisoning.Methods This study was divided into 2 parts.Part 1 was a population study:133 local residents exposed to airborne arsenic through coal-burning were selected as arsenic exposure groups including a non-patient group (25 cases),no obvious hepatopathy group (38 cases),mild (43 cases) and moderate to severe hepatopathy groups (27 cases) from areas affected by endemic arsenism in Guizhou Province.Thirty-four healthy residents from arsenic not affected areas were selected as controls.Peripheral blood samples were collected from all these people.The expression of TrxR1 mRNA was determined by real-time fluorescence quantitative PCR (qPCR),and enzyme activity of TrxR was tested by visible spectrophotometry.Part 2 was an animal experiment study:Thirty Wistar rats,weighing about 80-100 g,were divided into control group,drinking-waterborne arsenic poisoning group and coal-burning-borne arsenic poisoning group (including low,medium and high arsenic contaminated grain groups) by means of a table of random number according to body mass,6 rats in each group.The control group was fed with normal diet for 3 months; drinking-water-borne arsenic poisoning group and coal-burning-borne arsenic poisoning group were fed with 10 mg/kg As2O3 solution and different concentrations(25,50,100 mg/kg) of arsenic-containing feed,respectively,for 3 months.The expression of TrxR1 mRNA was determined by qPCR; protein expression level of TrxR1 in liver tissue was detected by immunohistochemistry,and enzyme activity of TrxR in serum and liver tissue was tested by visible spectrophotometry.Results The mRNA expressions of TrxR1 in peripheral blood were 1.599 8 (1.128 9-2.156 8),1.469 3 (1.146 1-1.976 3),1.203 6 (0.463 1-1.816 2) and 0.912 3(0.631 8-1.535 0),respectively,among non-patient group,no obvious hepatopathy group,mild and moderate to severe hepatopathy groups.Compared to the control group[1.649 7(1.161 1-2.380 2)],the differences were significant statistically in mild and moderate to severe hepatopathy groups (all P < 0.05).The enzyme activity of TrxR in peripheral blood was (3.12 ± 0.76),(2.81 ± 0.84),(2.52 ± 0.73),(2.42 ± 0.76)U/ml,respectively,in those corresponding groups.Compared to the control group [(3.02 ± 0.70)U/ml],the differences were significant statistically in mild and moderate to severe hepatopathy groups (all P < 0.05).The mRNA expressions of TrxR1 in peripheral blood were 1.05 ± 0.14,1.18 ± 0.18,1.04 ± 0.10 and 0.97 ± 0.13,respectively,among drinking-water-borne arsenic poisoning group,low,medium and high arsenic contaminated grain groups; all of which were lower than that in the control group (1.23 ± 0.15,all P < 0.05) except that of the low arsenic contaminated grain group.The mRNA expressions of TrxR1 in liver tissue were 0.78± 0.10,0.83 ± 0.10,0.79 ± 0.09 and 0.77 ± 0.11,respectively; all of which were lower than that in the control group (0.94 ± 0.12,all P < 0.05).The protein expression of TrxR1 in liver tissue was 310.33 ± 38.81,312.50 ± 23.36,305.67 ± 20.57 and 298.17 ± 23.52,respectively,among the arsenic poisoning groups; all of which were lower than that in the control group (348.50 ± 32.35,all P < 0.05).The enzyme activity of TrxR in serum was (4.22 ± 0.73),(4.86 ± 0.63),(4.04 ± 0.57),(3.73 ± 0.64)U/ml,respectively; all of which were lower than that in the control group [(9.52 ± 1.08)U/ml,all P < 0.05].The enzyme activity of TrxR in liver tissue was (14.82 ± 1.67),(18.76 ± 2.76),(14.90 ± 2.17),(11.55 ± 1.74) U/mg,respectively; all of which were lower than that in the control group [(23.71 ± 3.05)U/mg,all P < 0.05].Conclusion Arsenic aggravates liver injury of coal-burning arsenic poisoning through down-regulating the expressions of TrxR1 mRNA and protein and reducing its enzyme activity as well.
4.Effect of Ginkgo biloba on liver injury of arsenic poisoning rats caused by corn flour baked by high-arsenic coal
Maolin YAO ; Aihua ZHANG ; Chun YU ; Yuyan XU ; Yong HU
Chinese Journal of Endemiology 2017;36(5):333-337
Objective To explore the effects and the possible mechanism of Gingko biloba on liver injury due to arsenic poisoning in rats,and to provide experimental evidence for prevention and treatment of arsenic poisoning.Methods The corn powder baked by high arsenic coal was served as the main raw material to make feed containing arsenic.Forty healthy Wistar rats were randomly divided into 5 groups according to their body weights,including control group A,arsenic poisoning group,control group B,natural recovery group and Ginkgo biloba treatment group,eight rats in each group,half male and half female.The control group A rats were fed with normal diet ad libitum for 3.0 months;the arsenic poisoning group rats were freely given feed containing arsenic (100 mg/kg) for 3.0 months;the control group B rats were fed with normal diet ad libitum for 4.5 months;the natural recovery group rats were freely given arsenic (100 mg/kg) feed for 3.0 months,and then given a normal diet for 1.5 months;Ginkgo biloba treatment rats ingested arsenic feed for 3.0 months,and then give Ginkgo biloba solution (25 mg/kg) orally,6 d/week for 1.5 months,then back to normal diet.The content of arsenic in urine,liver,as well as the liver function indices [alanine aminotransferase (ALT),aspartate transaminase (AST),total bile acids (TBA),gamma glutamyl aminopeptidase (GGT),glutathione S-transferase (GSTs)] and the oxidative stress indexes [superoxide dismutase (SOD),glutathione peroxidase (GPx),thiol (-SH),malondialdehyde (MDA)] of liver homogenate,were measured.Results The arsenic content of urine and liver (geometric mean) of the rats in arsenic poisoning group (2 991.24 μg/g Cr,4.29 μg/g) were significantly higher than those in control group A (91.59 μg/g Cr,1.00 μg/g).Urinary arsenic and liver arsenic levels of rats in natural recovery and Ginkgo biloba treatment groups (467.39,334.48 μg/g Cr;,3.15,1.88 μg/g) were higher than those in control group B (99.54 μg/g Cr,0.85 μg/g).The arsenic contents of urine of the rats in natural recovery group,the arsenic contents of urine and liver of rats of Ginkgo biloba treatment group were all lower than those in arsenic poisoning group.The differences were significant (all P < 0.05).The activity/contents of AST,TBA,GGT,GSTs of rats in arsenic poisoning group [(212.88 ± 29.76) U/L,(19.19 ± 4.33) μmol/L,(1.73 ± 0.50) U/L,(196.21 ± 47.38) U/L] were all significantly higher than those in control group A [(142.63 ± 24.20) U/L,(6.23 ± 2.95) μmol/L,(0.77 ± 0.32) U/L,(142.86 ± 28.58) U/L].The activity/contents of TBA,GGT,GSTs in natural recovery group were (17.07 ± 3.92) μ,mol/L,(1.47 ± 0.57) U/L and (178.06 ± 27.37) U/L;and the contents of TBA in Ginkgo biloba treatment group were (13.60 ± 3.00) μmol/L;which were all higher than those in control group B [(7.55 ± 2.45) μmol/L,(0.74 ± 0.51) U/L,(145.17 ± 28.59) U/L].The activity of AST in natural recovery group [(137.44 ± 23.20) U/L],the activity/contents of AST,TBA,GGT and GSTs in Ginkgo biloba treatment group[(129.63 ± 31.25) U/L,(13.60 ± 3.00) μmol/L,(1.15 ± 0.48) U/L,(155.64 ± 20.79) U/L,respectively] were all lower than those in arsenic poisoning group.The content of TBA in Ginkgo biloba treatment group was lower than that of natural recovery group.The differences of those indexes were all significant (all P < 0.05).The activity/contents of SOD,GPx and-SH in arsenic poisoning group [(46.34 ± 11.39),(275.16 ± 92.00) U/mg prot and (0.08 ± 0.02) μmol/mg prot] were all significantly lower than those in control group A [(75.52 ± 8.72),(1 351.01 ± 395.96) U/mg prot,(0.13 ± 0.01) μmol/mg prot].The activity of SOD and GPx in natural recovery group [(42.44 ± 9.58),(694.87 ± 187.01) U/mg prot] were all lower than those in control group B [(68.17 ± 11.11),(1 342.80 ± 185.04) U/mg prot].The activity of GPx in natural recovery group,the activity/contents of SOD,GPx,-SH in Ginkgo biloba treatment group [(63.90 ± 10.44),(1 283.28 ± 373.87) U/mg prot,(0.12-± 0.02) μmol/mg prot] were all higher than those in arsenic poisoning group.The contents of SOD,GPx,-SH in Ginkgo biloba treatment group were higher than those of natural recovery group.The content of MDA in arsenic poisoning group [(3.05 ± 0.94) nmol/mg prot] was higher than that in control group A [(1.67 ± 0.55) nmol/mg prot].The content of MDA of rats in natural recovery and Ginkgo biloba treatment groups were (2.22 ± 0.93),(1.77 ± 0.37) nmol/mg prot,which were lower than those in the arsenic poisoning group.The differences of the above indexes were all significant (all P < 0.05).Conclusion Ginkgo biloba can reduce the accumulation of arsenic in the liver and ameliorate lipid peroxidation,relieve liver injury effectively in rats caused by coal-burning arsenic.
5.Isolation, culture and identification of pancreatic stem cells of Kunming mouse
Maolin QIN ; Jianjun LIU ; Zhongxiang YAO ; Deshan ZHOU ;
Journal of Third Military Medical University 1984;0(01):-
Objective To isolate and identify pancreatic stem cells of Kunming mouse and to observe the differentiation potency of those cells. Methods Pancreas of postnatal Kunming mice were digested by enzymes to isolate pancreatic stem cells. The potency of the cell differentiation was then identified with special marker of cytokeratin 19(CK 19), insulin and glucagons by immunocytochemical staining. Results Few epithelioid cells could be found to survive at the beginning of isolation, but when medium was replaced by that with growth factor, the cells proliferated quickly in fusiform shape and formed colony gradually. The cells were CK 19 immunoreactive positive after transfer of culture. After differentiation induced by high glucose, the cells formed the pancreatic islet like structures. Immunocytochemical staining showed that part of the cells of pancreatic islet like structures were insulin immunoreactive positive and few of the cells were glucagon immunoreactive positive. Conclusion Pancreatic stem cells of Kumming mouse can proliferate when cultured in vitro and have the potency of differentiation into ? and? cells of pancreatic islet.
6.Clinical application of the adjacent horn shaped perforator fasciocutaneous flap in the trunk area.
Wende YAO ; Xiaojing LI ; Jinlong NING ; Xinyi LI ; Zhao CHEN ; Maolin TANG ; Huairui CUI
Chinese Journal of Plastic Surgery 2014;30(4):241-244
OBJECTIVETo study the anatomy basis for the clinical application of the adjacent horn shaped perforator fasciocutaneous flap for the reconstruction of small and medium-sized defects in the trunk area.
METHODS(1) Ten adult antiseptic cadavers (20 sides) were perfused with red latex. The skin blood supply, line of the blood vessels, branches in accordance with the distribution and crossing were observed. (2) Fifteen cases with defects in the trunk were treated with the adjacent horn shaped perforator fasciocutaneous flaps. The defects size ranged from 5 cm x 5 cm to 13 cm x 13 cm with the size of the flaps ranging from 10 cm x 6 cm to 35 cm x 15 cm.
RESULTSThe trunk skin is supplied by mainly 17 groups arteries such as thyrocervical trunk, internal thoracic artery, posterior intercostal arteries, superior epigastric artery, arteria epigastrica inferior, lumbar arteries, and so on. The perforators (diameter > 0.5 mm) numbers are about 20, 40, 24, 6, on the chest, abdomen and perineum, upper back, waist, respectively. All the flaps survived completely with primary healing both on donor and recipient sites. The flaps color, texture, function and appearance were satisfactory during the follow-up period of 1-24 months.
CONCLUSIONSThe adjacent horn shaped perforator fasciocutaneous flap should be designed flexibly. The defects in the donor sites could be closed directly without skin graft. It is an effective, easy and ideal method for the reconstruction of large defects in the trunk.
Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Perforator Flap ; Skin Transplantation ; Torso ; surgery ; Young Adult
7.Transabdominal-transanal total mesorectal excision combined with liver metastasis resection for synchronous low rectal liver metastasis
Zhipeng ZHANG ; Maolin TIAN ; Chunhui YUAN ; Yimu JIA ; Hongwei YAO ; Dianrong XIU
Chinese Journal of Digestive Surgery 2016;15(2):123-127
Objective To investigate the application value of transabdominal-tansanal total mesorectal excision combined with liver metastasis resection for synchronous low rectal liver metastasis.Methods The retrospective descriptive study was adopted.The clinical data of a male patient with synchronous low rectal liver metastasis who was admitted to the Peking University Third Hospital in November 2015 was collected.Transabdominal-transanal total mesorectal excision combined with liver metastasis resection was performed after multidisciplinary team conference.The liver metastasis resection,vascular disconnection,lymph node dissection and upper and middle mesorectal disconnection were done by transabdominal approach.Then complete mesorectal excision and specimen removal of rectum and liver were done by transanal approach.The intraoperative status (operation method,operation time,volume of intraoperative blood loss,blood transfusion),occurrence of postoperative complications,results of pathological examination and follow-up were observed.The patient was followed up by outpatient examination till January 2016.Results The operation was performed successfully without severe perioperative complications.The operation time and volume of intraoperative blood loss were 360 minutes and 170 mL,respectively.The patient did not receive intraoperative blood transfusion,without urinary retention and presacral abscesses.The patient was discharged at postoperative day 9.The postoperative pathological results showed high-differentiated rectal protruded adenoma and high-middle differentiated adenocarcinoma metastasis in the liver tissue with the negative resection margins.The tumor sizes of rectum and liver metastasis were 5.0 cm × 5.0 cm× 1.5 cm and 1.5 cm × 1.0 cm × 1.5 cm,respectively.The tumor node metastasis (TNM) stage was stage Ⅳ (pT3N0M1).The patient had a good life quality during the follow-up of 1 month.Conclusion Transabdominal-transanal approach might provide an alternative operative approach and resection method for synchronous low rectal liver metastasis,with a good short-term outcome.
8.Biomarkers in rats for kidney damage characteristics of arsenism due to coal burning and benchmark dose analysis
Yuyan XU ; Aihua ZHANG ; Jun LI ; Liyuan CHEN ; Maolin YAO ; Chun YU ; Qibing ZENG ; Jiang HE
Chinese Journal of Pharmacology and Toxicology 2014;(2):243-247
OBJECTIVE Study the kidney toxic effects caused by burning coal endemic arsenism in rats,application bench mark dose (BMD) method to investigate the bench mark dose of urinary arsenic (UAs)and the changes in bio markers of renal function.METHODS Wistar rats were fed for 90 d with arsenic 0,25,50,100 mg·kg -1 conta minated feed.Urinary arsenic,kidney arsenic and renal function indicators were determined,and routine pathological and fibrosis of kidney were exa mined.UAs as the exposure bio marker,Uβ2-MG,UNAG and UALB for the effect bio markers,application bench mark dose method to calculate the BMD and BMDL of UAs for each effect bio markers.RESULTS UAs,KAs, Uβ2-MG,UNAG,UALB levels of rats in arsenic 100 mg·kg -1 group were increased than normal group (P <0.05);In light microscope,the results of HE staining of rat kidney in all arsenic dose groups showed infla mmatory cell infiltration,renal tubular epithelial cell swelling,renal interstitial capillary dila-tion,congestion and other varying degrees pathological changes,and the results of masson staining showed varying degrees of tubulointerstitial fibrosis;UAs as the exposure bio marker,Uβ2-MG,UNAG, UALB for the effects of mark,the BMD and BMDL of UAs for Uβ2-MG,UNAG,UALB were calculated, the BMD values were 998.9,1213.5,1386.9 μg·g -1 Cr,the BMDL values were 660.5,803.6 and 909. 4 μg·g -1 Cr,respectively.CONCLUSION Burning coal arsenic pollution can cause kidney da mage in rats,mini mal change nephropathy may be the pri mary pathological in the coal arsenic conta mination of kidney da mage.The BMD and BMDL of UAs were 998.9,660.5 μg·g -1 Cr,the early changes of renal function of burning coal arsenism in rats;it is reco mmended to use the more sensitive bio markers Uβ2-MG to calculate the biological exposure li mits on renal injury caused by arsenic.
9.Clinical efficacy of the laparoscope-assisted transanal total mesorectal excision for middle-low rectal cancer
Zhipeng ZHANG ; Hongwei YAO ; Ning CHEN ; Yang BAI ; Maolin TIAN ; Dechen WANG ; Jiong YUAN ; Dianrong XIU
Chinese Journal of Digestive Surgery 2017;16(7):695-700
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.