Objective:To investigate the effect of long-chain non coding RNA (lncrna) loc730101 in the proliferation, invasion and migration of U2OS cells, and its mechanism.Methods:From February, 2019 to October, 2019, U2OS cells cultured in vitro were divided into control group (normal culture), negative group (transfection nega- tive control), and interference group (transfection of interference sequences targeting LOC730101). The expression of LOC730101 in cells was detected by reverse transcription-polymerase chain reaction (RT-PCR). Cell proliferation ac tivity was tested by methylthiazolyldiphenyl-tetrazolium bromide (MTT) method. Cell clone formation rate was mearused by plate clone formation test. Cell cycle distribution was tested by flow cytometry. Cell invasion and migra- tion were examed by Transwell chamber. The expression levels of epithelial-mesenchymal transition-related proteins Vimentin, N-cadherin, E-cadherin, and Wnt/β-catenin signaling pathway related proteins in cells β-catenin, c-Myc, cyclin D1(CyclinD1) and matrix metalloproteinase-7(MMP-7) proteins were detected by Western blotting method. The date was statistical analysed and considered as statistically significant when P<0.05. Results:Compared with the control group, the expression level of LOC730101 (0.25±0.03 and 1.00±0.06) in interference group and control group, respectively. The same below), cell survival rate [(57.65±3.26)% and (100.00±7.39)%], clone formation rate [(13.03± 2.12)% and (25.35±3.58)%], number of invasive cells(51.36±3.48 and 92.85±6.62), number of migrating cells (77.15± 5.05 and 136.92±15.35), the percentage of cells in S phase [(20.54±2.15)% and (28.15±2.38)%] and G 2/M phase [(16.87±2.12)% and (23.36±3.12)%], as well as the expression level of Vimentin (0.52±0.04 and 1.17±0.13), N-cadherin (0.31±0.03 and 0.65±0.04), β-catenin (0.42±0.03 and 0.73±0.04), c-Myc (0.29±0.03 and 0.65±0.03), CyclinD1 (0.26± 0.02 and 0.58±0.04), MMP-7 protein (0.55±0.03 and 0.86±0.06) was decreased significantly ( P<0.05), while the per- centage of cells in G 0/G 1 phase [(62.62±5.15)% and (48.46±3.65)%] and the expression level of E-cadherin protein(0.82± 0.06 and 0.38±0.03) were increased significantly in the interference group ( P<0.05). But there was no significant differ- ence in each index in the negative group ( P>0.05). Conclusion:Reduce the regulation of LOC730101 can inhibit the proliferation, invasion and migration of U2OS cells, and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway.

Objective:To evaluate the personalized 3D printed guide template used in the osteotomy for malunion of tibial fracture.Methods:A retrospective analysis was conducted of the 30 patients who had been treated for malunion of tibial fracture at Department of Orthopaedics, The First Affiliated Hospital to Zhengzhou University from January 2010 to January 2018. Of them, 15 used a personalized 3D printed guide template in the osteotomy (3D printing group). They were 9 males and 6 females, with an age of 46.3 year±8.2 years. The fracture malunion was located in the upper and middle tibia in 11 cases, in the lower tibia in 4 cases, on the left side in 6 cases and on the right side in 9 ones. There were 8 cases of varus deformity and 7 ones of valgus deformity. Their preoperative fracture deformity angle was 24.3°±5.5°. The other 15 patients were treated with conventional surgery (conventional group). They were 10 males and 5 females, with an age of 47.1 years±6.0 years. The fracture was located in the upper and middle tibia in 12 cases, in the lower tibia in 3 cases, on the left side in 5 cases and on right side in 10 cases. There were 7 cases of varus deformity and 8 ones of valgus deformity. Their preoperative fracture deformity angle was 22.5°±5.4°. The 2 groups were compared in terms of preoperative baseline data, operation time, intraoperative blood loss and postoperative recovery of the alignment of lower limb.Results:There were no significant differences in the preoperative baseline data between the 2 groups, showing comparability ( P>0.05). The 3D printing group was followed up for an average of 12 months while the conventional group for an average of 10 months. The operation time for the 3D printing group was significantly shorter than that for the conventional group(102.2 min±13.0 min versus 137.9 min ±10.5 min), the intraoperative blood loss for the former significantly less than that for the latter (77.3 mL ± 39.7 mL versus 163.3 mL ± 35.2 mL), and the postoperative malunion angle in the former significantly smaller than that in the latter (1.9°±0.4° versus 3.2°±0.9°) (all P< 0.05). The last follow-ups revealed no implant failure or re-malunion but fine healing of the osteotomy sites and good recovery of the alignment of lower limb in the 2 groups. Conclusion:A personalized 3D printed guide template used in the osteotomy for malunion of tibial fracture is an effective aid because it can facilitate precise osteotomy, reduce operation time and intraoperative blood loss and help correct the alignment of lower limb, leading to good short-term surgical outcomes.

Objective: To describe the general and histological features of the full-length superficial fascia of the circumferential upper limb. Methods: Fresh frozen arm specimens were dissected, and then MRI imaging in vivo, enhanced CT angiography and HE histological staining were used to describe the characteristics of the full-length superficial fascia of the circumferential arm and its relationship with important blood vessels. Results: The four typical structures of the superficial fascia of the arm were divided into subcutaneous superficial fat, membrane-like substance, deep fat and deep fascia from superficial to deep. The thickness and stratification, fusion degree and histological characteristics of the superficial fascia of these four layers were obviously different in different levels and regions of the arm. MRI confirmed that the total thickness of superficial fascia gradually decreased from shoulder to wrist. Venography showed that the cephalic vein ran below the second layer of superficial fascia and above the deep fascia. The basilic vein originated from the dorsal vein network of the hand and always lied below the second layer of membranous material until the basilic vein penetrates below the deep fascia of the upper arm. Conclusions The deep understanding of the circumferential full-length of superficial fascia structure of the upper limb provides an important theoretical basis for improving the surgical safety and fine operation for the Dynamic Arm Circumferential Liposuction.

Objective: To describe an extended whole upper limb liposuction technique and evaluate its clinical effect. Methods: 34 patients who underwent upper limb liposuction from February 2018 to February 2019 were selected and the clinical data were retrospectively summarized. Patients were treated with upper arm ring/upper arm ring + forearm 1/3 rings/whole arm ring aspiration combined with accessory mammary gland, armpit, scapula and other adjacent aesthetic parts of extended liposuction. The preoperative and postoperative maximum circumference, thickness of the anterior and posterior subcutaneous tissue and skin laxity were measured and compared. analysis of complications, and evaluation of patients′ satisfaction through satisfaction questionnaire. Results: The arm shape of all patients was significantly improved, the maximum circumference of the arm was reduced (16.2±4.0)%, the distance of the upper arm was reduced (29.5±8.9)%, the thickness of the posterior subcutaneous tissue was reduced (56.6 ±6.2)%, and the thickness of the anterior subcutaneous tissue was reduced (44.7±9.6)%. There were three cases of mild anemia after operation, and no other serious complications occurred. Patient satisfaction was very high. Conclusions This is an innovative arm liposuction technique with hidden incision. The effect of improvement is remarkable. The patient′s satisfaction is high.

Objective: To explore the clinical effect and safety of minor liver resection for hilar cholangiocarcinoma (HC) of Bismuth-Corlette type Ⅲ and Ⅳ. Methods: From May 2007 to May 2017, the clinical data of 108 patients with Bismuth-Corlette type Ⅲ and Ⅳ HC underwent hepatectomy were collected and analyzed retrospectively.There were 56 males and 52 females, aged (57.2±5.3) years (ranged 48-76 years) .Among the 108 cases, there were 51 cases of type Ⅲa, 40 cases of type Ⅲb and 17 cases of type Ⅳ. Small-scale hepatectomy (≤3 hepatectomy) was performed in 70 cases, including 8 cases of 4b segment resection, 28 cases of 4b segment+5 segment resection, and 34 cases of partial 4 segment+partial 7 segment+partial 1 segment resection. Large-scale hepatectomy was performed in 38 cases (>3 segments) , of which 30 cases were treated with 2 segments+3 segments+4 segments+1 segment, and 8 cases were treated with 5 segments+7 segments+8 segments+1 segment. t′ test was used to analyze the data which did not conform to the normal distribution, and χ² test was used to calculate the incidence of postoperative complications and the 1, 3, and 5-year cumulative overall survival rate. Results: (1) The operation time of minor liver resection group ((180±25)minutes) was shorter than that of major liver resection group ((210±35)minutes) (t′=4.676, P<0.05) , the amount of blooding operation time of minor liver resection group ((310±80)ml) was less than that of major liver resection group ((500±110)ml)in the operation (t′=9.385, P<0.05) , and the difference was statistically significant. (2) The incidence of complications was lower in minor liver resection group and major liver resection group, and the difference was statistically significant (χ²=5.230, P<0.05) . (3) The actual 1-, 3- and 5-year survival rates were 87.1%, 58.4%, 30.0% and 84.2%, 57.9%,31.6%, respectively. There were no significant differences in survival rates in two groups in 1-, 3- and 5-year survival rates (χ²=0.177, P=0.674; χ²=0.005, P=0.946; χ²=0.029, P=0.865) . Conclusions Compared to patients with major liver resection, Minor liver resection for selected patients with HC of Bismuth-Corlette Ⅲ and Ⅳaccording to our criteria achieved better long-term outcomes. Chen′s biliojejunostomy is a simple, effective and safe method, which can be widely used when there are multiple biliary intestinal anastomosese.

Objective To investigate the association of serum brain-derived neurotrophic factor (BDNF) and methylglyoxal (MG) levels with cognitive function in elderly patients with type 2 diabetes mellitus (T2DM). Methods The normal population and elderly patients with T2DM were frequency-matched by age, sex, and educational level. BDNF was detected by ELISA assay, MG by HPLC assay, and cognitive function by sets of repetitive mental state examination (RBANS) in the two groups. Results (1) Compared with control group, serum BDNF level in T2DM group was significantly decreased [ (4.97±3.05 vs 11.77±7.92)ng /ml, P<0.01]while serum MG level was elevated [(67.91 vs 43.86) nmol /L, P<0.05]. The increasing of serum MG was related to the decreasing of serum BDNF. (2) Compared with control group, the scores for standardized tests of cognitive scale, visual breadth, immediate memory, delayed memory, and attention areas in T2DM group were significantly decreased (all P<0.05). After the influencing factors were adjusted by multiple regression, the associations of serum BDNF level with cognitive scale standardized score, the delay associated with memory and attention functions were still evident, and serum MG level in T2DM group was still related with the levels of delayed memory, immediate memory, total scale standardization (all P<0.05). (3) Serum BDNF level was negatively correlated with serum MG level (P=0.031). Conclusions Cognitive function of elderly patients with T2DM is related with serum MG and BDNF levels. The increased serum MG as well as the decreasd serum BDNF levels maybe involved in the pathogenesis of impaired cognitive function.

Cell biology is a highly experimental discipline, and is a discipline with theoretical knowledge closely combined with practical operation.Most of the medical graduate students should master the cell biology theory in the graduate stage, and get on a series of cell biology researches.Therefore, it is important to master the most basic cell culture techniques.As a technical instructor in the cell culture laboratory, I have been summarizing and optimizing the teaching content and teaching mode during the past few years.The aim of the teaching work is to train the students' scientific attitude of seeking truth from facts so as to develop good experimental habits and to master the basic skills of cell culture.

OBJECTIVETo construct and characterize the TGF-β1, induced epithelial-mesenchymal transition (EMT) model of keloid epithelial cells in vitro, and to investigate the expression of epithelial stem cells related surface markers in keloid epithelial cells during EMT induction.

METHODSThe epithelial cells from 3 keloid samples of ears were cultured in vitro and induced by transforming growth factor betal (TGF-β1, 1 ng/ml) for 5 days, which was the experimental group, the same cells untreated were considered as the negative control group. The expressions of EMT-associated markers and regulative genes were detected using immunofluorescence staining, real-time PCR and western blot analysis. Then the surface markers of epithelial stem cells were detected using real-time PCR. Statistical significance was determined using Independent-Samples t Test, a p value less than 0. 05 was considered statistically significant.

RESULTSThe mRNA expression of transcription factor snail2 and mesenchymal-specific marker vimentin increased significantly in TGF-β1, induced keloid epithelial cells (P < 0. 05), in which snail2 increasing from 0. 91 ± 0. 23 to 1. 69 ± 0. 10, and vimentin from 5. 86 ± 2. 07 to 24. 29 ± 5. 39. Whereas the mRNA expression of epithelial-specific marker E-cadherin decreased from 1. 06 ± 0. 19 to 0. 65 ± 0. 09. The mRNA expression of CD29 and Lgr6, two surface markers of epithelial stem cells, significantly increased after induction of the TGF-β1, (P < 0. 05), from 0. 55 ± 0. 14 and 1. 61 ± 0. 31 to 1. 19 ± 0. 12 and 3. 84 t 0. 62 respectively. In induced cells, the immunofluorescence results showed staining of E- cadherin became faint, but the number of positive staining cells of vimentin increased. Western blot confirmed the protein expression of E-cadherin weakened, and the vimentin and p-Smad3 enhanced (P < 0. 05).

CONCLUSIONSTGF-β1, initiated EMT in keloid epithelial cells by inducing the up-regulation of snail2, and TGF-β1,/Smad3 signaling pathway was involved in EMT. EMT could change the phenotype of epithelial stem cells in keloid.

Biomarkers ; metabolism ; Cadherins ; genetics ; metabolism ; Epithelial Cells ; drug effects ; physiology ; Epithelial-Mesenchymal Transition ; drug effects ; physiology ; Humans ; In Vitro Techniques ; Keloid ; pathology ; RNA, Messenger ; metabolism ; Signal Transduction ; Smad3 Protein ; genetics ; metabolism ; Snail Family Transcription Factors ; Transcription Factors ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; pharmacology ; Up-Regulation ; Vimentin ; genetics ; metabolism

Objective To investigate the role of human membrane associated sialidase (Neu3) in multidrug resistance ( MDR) of K562 cells. Methods K562 cells and K562/ADM cells were treated with DNR alone or with combination of DNR and NeuAC2en. The cell survival rate was measured by MTT assay; the expression of MDR related factors and apoptosis related factors were determined by Western blot and RT-PCR; Neu3 activity was detected by TBA reaction. Results The survival rate of K562/ADM cells was higher than that of K562 cells; NeuAC2en showed synergistic effect with DNR(P <0. 01) ; Neu3 activity of K562 and K562/ADM cells decreased after DNR or NeuAC2en induction and the decrease was most significant in the combination treatment group (P <0. 01). Under the identical condition, the protein expression of P-gp and Neu3 and the mRNA expression of MDR1, Neu3, BCL-xl and BCL-2 increased comparing with K562 cells. The mRNA level of BAX in K562 and K562/ADM groupsdid not changed significantly. MDR1, Neu3,BCL-xl and BCL-2 decreased after DNR induction and down-regulated most obviously in the groups treated by DNR combined with NeuAC2en. Conclusion Neu3 may be related with multidrug resistance of K562/ADM cell through regulating apoptosis and the expression of MDR1.

Objective To explore the feasibility of using human umbilical cord derived mesenchymal stem cells as seed cells to repair sciatic nerve defects of rats by tissue engineering methods. Methods Mesenchymal stem cells from human umbilical cord were cultured and induced into neuron-liked cells,which were co-cultured with acellular basal lamina tube to construct tissue engineering nerve;models of sciatic nerve defects 10 mm in length were set up with thirty healthy adult SD rats and were divided randomly into 3 groups:tissue engineering nerve group (group A, compound of human umbilical cord derived mesenchymal stem cells and acellular basal lamina tube), pure acellular basal lamina tube group (group B), and autogenous nerve bridging group (group C). Evaluation of electrophysiological and histological results was carried out 10 weeks after operation. Results The engineering nerve group had good result in nerve regeneration which was close to the effect of autogenous nerve transfer group (group A), and much better than the effect of pure acellular basal lamina tube group. Conclusion Engineering nerves from human umbilical cord derived mesenchymal stem cells can effectively repair 10 mm defects of sciatic nerve.