Although somatic cells can be reprogrammed to pluripotent stem cells (PSCs) with pure chemicals, authentic pluripotency of chemically induced pluripotent stem cells (CiPSCs) has never been achieved through tetraploid complementation assay. Spontaneous reprogramming of spermatogonial stem cells (SSCs) was another non-transgenic way to obtain PSCs, but this process lacks mechanistic explanation. Here, we reconstructed the trajectory of mouse SSC reprogramming and developed a five-chemical combination, boosting the reprogramming efficiency by nearly 80- to 100-folds. More importantly, chemical induced germline-derived PSCs (5C-gPSCs), but not gPSCs and chemical induced pluripotent stem cells, had authentic pluripotency, as determined by tetraploid complementation. Mechanistically, SSCs traversed through an inverted pathway of in vivo germ cell development, exhibiting the expression signatures and DNA methylation dynamics from spermatogonia to primordial germ cells and further to epiblasts. Besides, SSC-specific imprinting control regions switched from biallelic methylated states to monoallelic methylated states by imprinting demethylation and then re-methylation on one of the two alleles in 5C-gPSCs, which was apparently distinct with the imprinting reprogramming in vivo as DNA methylation simultaneously occurred on both alleles. Our work sheds light on the unique regulatory network underpinning SSC reprogramming, providing insights to understand generic mechanisms for cell-fate decision and epigenetic-related disorders in regenerative medicine.
Male ; Mice ; Animals ; Cellular Reprogramming/genetics* ; Tetraploidy ; Pluripotent Stem Cells/metabolism* ; Induced Pluripotent Stem Cells/metabolism* ; DNA Methylation ; Spermatogonia/metabolism* ; Germ Cells/metabolism*

Enhanced recovery after surgery (ERAS) represents a transformation to the evidence-based medicine paradigm in perioperative treatment. ERAS aims to reduce recovery time, postoperative complications, and healthcare costs. Here, we review the development and principal components of ERAS along with its application in abdominal tumor surgery. The benefits and feasibility of the ERAS protocol have been demonstrated in clinical practice, particularly in relation to colorectal surgery.

Objective To evaluate the clinical effect of iliac artery interventional chemotherapy combination with bladder perfusion for high level of bladder urothelial carcinoma after transurethral bladder tumor electricity cut operation for those who intend to retain the bladder.Methods From February 2010 to December 2016,a total of 74 cases high-grade urothelial carcinoma of the bladder,giving chemotherapy and suitted support were retrospectively analyzed.The patients were divided into two groups according to different chemotherapy methods.Artery perfusion chemotherapy group:43 cases treated by transurethral bladder tumor cutting + bilateral iliac artery interventional perfusion chemotherapy + bladder perfusion in combination therapy;Intravenous chemotherapy group:31 cases underwent transurethral bladder tumor cut method + cisplatin,vein gemcitabine + bladder perfusion chemotherapy combined therapy.According to the ((x) ± s) between the groups.The method of count data is represented by chi-square test.Kaplan-meier method was used to describe the survival time of two groups of patients after operation.Results Two groups of patients with postoperative there was no statistically significant difference in disease-free survival time.In the bone marrow suppression (x2 =4.956,P =0.029);gastrointestinal tract reaction (x2 =5.912,P =0.012);dermatitis,mucositis (x2 =4.276,P =0.013),etc.,has the advantages in the iliac artery interventional perfusion chemotherapy (P ＜ 0.05).Conclusion Via efficient iliac arterial infusion chemotherapy in patients with venous chemotherapy patients,to the retention of the bladder has a certain value,and the incidence of adverse reactions after surgery is low,have certain advantages.

Objective To establishing an immortal cell line of familial papillary thyroid carcinoma (FPTC), and explore a new approach for studying familial non-medullary thyroid carcinoma (FNMTC). Methods The specimen from a patient with FPTC was selected, separated, and the primary cells were cultured using DMEM/F12 medium (with TSH, T3, EGF and hydrocortisone). To inducing cell immortalization, the exogenous genes SV40T/TERT were transfected into cells by two ways. RT-PCR was used to detect the expressions of thyroid peroxidase (TPO), thyroid globulin (TG), thyroid stimulating hormone receptor (TSHR) and sodium/iodide co-transporter (NIS). Immunofluorescence method was used to detect the expressions of TPO and GPC3. In order to detect the genomic mutations, the peripheral blood DNA of the patient was extracted. The cell genome was detected. Results The FPTC cells adhered to the plate and showed an irregular polygon shape. The cells can stably grow for six months, FPTC-S (with SV40T transfected) passaged to p26, FPTC cells passaged to p23 and FPTC-ST (with SV40T/TERT transfected) passaged to p19. Both FPTC-S and FPTC-ST can stably express TPO, TG and TSHR in mRNA level. MLH1 R217C mutation existed in the peripheral blood of the patient, and BRAF V600E mutation existed in the primary cultured cells. Either the primary or the immortal cells showed MLH1 R217C mutation. Conclusion This study preliminarily established an immortal cell line of familial papillary thyroid carcinoma with MLH 1 R217C and BRAF V600E mutations. This cell line provides a research model for studying these mutations in FPTC.

Objective To observe the effect of mesenchymal stem cells ( MSC) in treating ischemic diabetic ulcers, and to explore its clinical perspectives. Methods Prepared electro-spinning biomaterials and cultured MSC to study effect of MSC composite biomaterials in vitro by scanning electron microscope,MTT array and influence of MSC conditioned medium on endothelial cells. The use of 5 to 8 week old male C57BL/6J mice were prepared into diabetic mice,femoral artery ligation in the proximal thigh,in dorsal skin full-thickness wounds caused by the diameter 5 mm. Then the effect of MSC composite biomaterials on ischemic diabetic ulcers was determined. Results This study found that the MSC grow well on electro-spinning biomaterials. Cells foot extension and connections between cells were observed by scanning electron microscope. Stimulated by high glucose,growth and proliferation of MSC has a stronger ability on biomaterials. MSC condi-tioned medium on biomaterials increased human umbilical vein endothelial cells proliferation ability. Conclusion MSC composite biomateri-als can effectively improve the treatment effect of MSC on ischemic diabetic ulcers. The study indicated stem cells composite biomaterials have great potential and application prospect in the treatment of ischemic diabetic ulcers healing.

Objective To study the feasibility of model building methods by analyzing the advantages and disadvantages between the two different methods to build off-pump coronary artery bypass graft animal model. Methods Twenty dogs were randomly divided into two groups:brachiocephalic artery group and descending aorta group. Small-caliber heterogeneous vascular vessels were used as bridge vessels. The incision was in the fourth intercostal space of the left chest. Vascular anastomosis was firstly done between the brachiocephalic artery and bridge vessels,or between descending aorta and bridge vessels,prior to coronary vascular and bridge vessels anastomosis. Results The dogs of two groups were not dead during operation. Brachiocephalic artery group and the descending aorta group:aortic vascular anastomosis times were (33.9 ±4.8) min and (29.6 ±3.5) min respectively (P<0.05),coronary vascular anastomosis time were (28.5 ±3.0) min and (28. 1 ± 2. 3) min respectively (P>0. 05). The surgical blood losses of the two groups were (77. 5 ± 16. 2) mL and (66. 5 ± 12. 3) mL re-spectively (P>0. 05). After side clamping descending aorta,femoral blood pressure significantly decreased in descending aorta group,and the two dogs had melena after operation. Conclusion Off-pump coronary artery bypass graft models were both constructed successfully by the two ways. Descending aorta group of femoral artery blood pressure violently fluctuated and had abdominal organs’ ischemia reperfusion in-jury. Though brachiocephalic artery group anastomosis group spent a little longer time,they had stable artery blood pressure during operation. As a result,the way of constructing animal model of brachiocephalic artery group is safer.

Objective To evaluate the clinical effect of InterTAN (IT) and dynamic hip screw (DHS) in treatment of femoral intertrochanteric fractures.Methods From July 2008 to November 2011,the patients with femoral intertrochanteric fractures were treated by IT (IT group,21 patients) and DHS (DHS group,35 patients).The intraoperative data (operative time,intraoperative blood loss,incision length and hospitalization period),postoperative complications (wound infection,lower limb deep venous thrombosis,bedsore,urinary tract infection and pneumonia) and fracture healing (fracture healing time,femoral neck crispation,coxa vara and Harris scores) were compared.Results IT group was superior to DHS group in operative time,intraoperative blood loss,incision length and hospitalization period [(62.16 ± 15.26) min vs.(85.78 ± 19.50) min,(237.73 ± 108.51) ml vs.(490.87 ± 127.90) ml,(10.20 ±2.02) cm vs.(19.83 ± 2.98) cm and (10.23 ±5.52) d vs.(15.58 ±5.32) d,P ＜0.01].However,there was no significant difference in postoperative complications (P ＞ 0.05).There was significant difference between IT group and DHS group in femoral neck crispation,coxa vara [9.52％ (2/21)vs.28.57％ (10/35),14.29％ (3/21)vs.31.43％ (11/35),P ＜ 0.01].There was no significant difference in fracture healing time and Harris scores (P ＞ 0.05).Conclusions Compared with DHS,IT has extensive application,shorter surgical time and reliable fixation effect.Therefore,IT is suitable to femoral intertrochanteric fractures.

[Objective]To evaluate the effects of Austin surgery for mild and moderate juvenile hallux valgus. [Method]From November 2005 to Januuary 2007,16 juvenile hallux valgus patients(28 feet)with average of 17.8 years(range,17 to 19 years)were treated with Austin surgery.The X-ray films of all patients were obtained before operation,and twelve months,fifteen months after operation.The hallus valgus angle,intermetatarsal angle,proximal articular set angle,and distal articular set angle were measured and analyzed on radiographs.The surgical outcome was evaluated combined with the Gu Xiang-jie's score.All pamameters were statistically analyzed. [Result]All the patients were completely followed up.Hallax valgus angle reduced from 26.3?1.19 to 11.7?0.40,intermetatarsal angle reduced from 14.1?0.82 to 7.2?0.85,proximal articular set angle from 12.7?0.28 to 6.4?0.54(all P

Objective To provide experimental data by exploring the anatomic structure of adult corpus penis vein and lymphatic vessel and the length after releasing penile superficial ligament. Methods The penis length of 16 adult corpuses was measured immediately after releasing the superficial ligament, and make a statistical analysis between the prolonged portion and corpus height. A total of 40 cases of fresh penis specimens were divided averagely into two groups which were performed by vascular casting, indirect-injection of lymphatic vessel, shiff stating and dissection to observe their anatomic structure. Results Prolonged penis length ranged from 2-3 cm. Indian ink of 10% injected from penile glans and frenulum of prepuce mainly flew back through dorsal and lateral lymphatic vessel, but not through ventral vessel. Conclusion Penis can be prolonged about 2-3 cm by releasing the superficial ligament out of erectile condition. One-third proximal lateral area is advised for injection of bulbocavernosus body because blood vessels are less here. Obstinate edema after penis lengthening surgery has a close relation with the damnification of dorsal superficial vein and dorsal and lateral lymphatic vessel of penis.

OBJECTIVETo explore the diagnostic value of MRI and SPECT in early postburn brain edema in severely burned dogs.

METHODSTwenty-six mongrel dogs were randomized into control (n = 6) and burned groups in which every 5 dogs were allotted to each of following time points: 6, 12, 18 and 24 postburn hours (PBHs). The dogs in burn groups were inflicted with 50% TBSA of III degree skin burn and were infused with 5% glucose solution after 6 PBHs, so that severe early postburn brain edema was produced. MRI and SPECT were employed to observe dynamically the brain of dogs in all groups. The results were collected and compared with one another.

RESULTSThe results indicated that with MRI brain morphological change of early brain edema could be shown as early as within 12 PBH and diffuse brain edema became more obvious with elapse of time. The changes might be difficult to be found by MRI when T(1)WISIR decreased below 10%. T(2)WI SIR increased by 8.29% at 24 PBH with blurred demarcation between the brain gray and white matters. There was diffused and progressive nuclide ((99)TCm-ECD) concentration in the brain tissue as shown by SPECT at 6 PBH. The radio-nuclide taking ratio increased significantly after 12 PBH, especially at 24 PBH (P < 0.01) when compared with that before burn.

CONCLUSIONCombined application of MRI and SPECT could evidently increase sensitivity and specificity in the diagnosis of early postburn brain edema.

Animals ; Brain Edema ; diagnosis ; diagnostic imaging ; Burns ; complications ; Dogs ; Magnetic Resonance Imaging ; Male ; Tomography, Emission-Computed, Single-Photon