Mechanism of Qizhu Kang'ai Prescription for Inhibiting Proliferation of Hepatocellular Carcinoma by Regulating Tumor Metabolic Reprogramming via PCK1/Akt/p21 Signal Axis / 中国实验方剂学杂志

ObjectiveTo study the effect of Qizhu Kang'ai prescription （QZAP） on the gluconeogenesis enzyme phosphoenolpyruvate carboxykinase 1 （PCK1） in the liver of mouse model of liver cancer induced by diethylnitrosamine （DEN） combined with carbon tetrachloride （CCl4） and Huh7 cells of human liver cancer， so as to explore the mechanism on regulating metabolic reprogramming and inhibiting cell proliferation of liver cancer cells. MethodDEN combined with CCl4 was used to construct a mouse model of liver cancer via intraperitoneal injection. A normal group， a model group， and a QZAP group were set up， in which QZAP （3.51 g·kg-1） or an equal volume of normal saline was administered daily by gavage， respectively. Serum and liver samples were collected after eight weeks of intervention. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， γ-glutamyltransferase （γ-GT）， and alpha-fetoprotein （AFP） in mice were detected to evaluate liver function changes of mice in each group. Hematoxylin-eosin （HE） staining and Sirius red staining were used to observe pathological changes in liver tissue. In the cell experiment， Huh7 cells were divided into blank group， QZAP low， medium， and high dose groups and/or PCK1 inhibitor （SKF-34288 hydrochloride） group， and Sorafenib group. The corresponding drug-containing serum and drug treatment were given， respectively. Cell counting kit-8 （CCK-8） method， colony formation experiment， Edu fluorescent labeling detection， intracellular adenosine triphosphate （ATP） content detection， and cell cycle flow cytometry detection were used to evaluate the proliferation ability， energy metabolism changes， and change in the cell cycle of Huh7 cells in each group. Western blot was used to detect the protein expression levels of PCK1， serine/threonine kinase （Akt）， phosphorylated Akt （p-Akt）， and cell cycle-dependent protein kinase inhibitor 1A （p21）. ResultCompared with the model group， the pathological changes such as cell atypia， necrosis， and collagen fiber deposition in liver cancer tissue of mice in the QZAP group were alleviated， and the number of liver tumors was reduced （P<0.01）. The serum ALT， AST， γ-GT， and AFP levels were reduced （P<0.01）. At the cell level， compared with the blank group， low， medium， and high-dose groups of QZAP-containing serum and the Sorafenib group could significantly reduce the survival rate of Huh7 cells （P<0.01） and the number of positive cells with Edu labeling （P<0.01） and inhibit clonal proliferation ability （P<0.01）. The QZAP groups could also reduce the intracellular ATP content （P<0.05） and increase the distribution ratio of the G0/G1 phase of the cell cycle （P<0.05） in a dose-dependent manner. Compared with the model group and blank group， PCK1 and p21 protein levels of mouse liver cancer tissue and Huh7 cells in the QZAP groups were significantly reduced （P<0.05，P<0.01）， and the p-Akt protein level was significantly increased （P<0.01）. Compared with the blank group， the ATP content and cell survival rate of Huh7 cells in the SKF-34288 hydrochloride group were significantly increased （P<0.05）， but there was no statistical difference in the ratio of Edu-positive cells and the proportion of G0/G1 phase distribution. Compared with the SKF-34288 hydrochloride group， the QZAP combined with the SKF-34288 hydrochloride group significantly reduced the ATP content， cell survival rate， and Edu-positive cell ratio of Huh7 cells （P<0.05） and significantly increased the G0/G1 phase distribution proportion （P<0.05）. ConclusionQZAP may induce the metabolic reprogramming of liver cancer cells by activating PCK1 to promote Akt/p21-mediated tumor suppression， thereby exerting an anti-hepatocellular carcinoma proliferation mechanism.

High preoperative neutrophile-lymphocyte ratio predicts poor prognosis of patients undergoing radical cystectomy for nonurothelial carcinoma of the bladder / 现代泌尿外科杂志

【Objective】 To investigate the predictive value of high preoperative neutrophile-lymphocyte ratio (NLR) for the prognosis of nonurothelial carcinoma of the bladder (NUBC) after radical cystectomy (RC). 【Methods】 Clinical and follow-up data of NUBC patients undergoing RC during Jan.2005 and Dec.2020 were collected. The optimal cut-off value of NLR was determined with the receiver operating characteristic (ROC) curve. The survival curve was drawn with Kaplan-Meier method to compare the differences in cancer specific survival (CSS) and overall survival (OS) between the high-NLR and low-NLR groups. The independent risk factors of CSS and OS were screened with Cox proportional hazard regression model. 【Results】 Of the 62 eligible cases,34 (54.8%) were diagnosed with adenocarcinoma,17 (27.4%) with squamous cell carcinoma, 6 (9.7%) with small cell carcinoma and 5 (8.1%) with sarcoma. Kaplan-Meier analysis results showed high NLR was associated with poor CSS (P=0.001) and OS (P<0.001). Cox regression results indicated that high NLR (HR=2.42, 95%CI: 1.12-5.23, P=0.025) and advanced pathologic tumor stage (HR=3.21, 95%CI:1.53-6.74,P=0.002) were independent risk factors of unfavorable CSS. Similarly, high NLR (HR=2.75, 95%CI: 1.35-5.56, P=0.005) and advanced pathologic tumor stage (HR=2.81, 95%CI:1.43-5.57, P=0.003) were independent risk factors of unfavorable OS. 【Conclusion】 As an independent risk factor of unfavorable CSS and OS in NUBC patients undergoing RC, high preoperative NLR is of great value in the prediction of long-term prognosis and may help to optimize individualized treatment.

Study on the biomechanical analysis and influencing factors study for the treatment of humeral epicondylitis by Bachuorounian manipulation / 中国骨伤

OBJECTIVE@#To quantitatively study the biomechanical parameters of Bachuorounian manipulation in the treatment of humeral epicondylitis, and discuss the effects of individual characteristics on the biomechanical parameters were discussed.@*METHODS@#From July 2019 to February 2020, 40 patients with external humeral epicondylitis were selected, including 18 males and 22 females, ranging in age from 20 to 50 years old, with an average of (34.37±8.41) years old;and the course of disease ranged from 1 to 11 months, with a mean of (6.05±2.71) months. The biomechanical parameters of the elbow joint of the affected side were measured by using the biomechanical sensor. At thesame time, the individual characteristic parameters of patients were collected to analyze the influence of different individual characteristics of patients on biomechanical parameters.@*RESULTS@#The results of mechanical analysis in each stage of the bachuorounian manipulation were as follows:the rolling back rotation force was (31.17±2.99) N;the buckling bending drawing force was (44.99±2.38) N;the rolling pre rotation force was (31.03±2.75) N;and stretching drawing force was (48.75±2.09) N. The correlation analysis between the parameters showed that there was a significant positive correlation between the buckling bending drawing force and the stretching drawing force parameters, and a significant positive correlation between the rolling force back-rotation force and the rolling pre-rotation force parameters. The multivariate linear regression analysis on the parameters of influencing factors and manipulative biomechanics showed that there was a significant correlation between body weight and rolling back-rotation force, significant correlation between elbow tenderness and the buckling bending drawing force, and significant correlation between disease duration and the stretching drawing force.@*CONCLUSION@#The bachuorounian manipulation of humeral epicondylitis has a certain range of operating force. Manipulation of each stage has a correlation and systematic. The patient's weight, elbow tenderness and disease course are important factors affecting the bachuorounian manipulation.

Minimally invasive direct lateral approach debridement, interbody bone grafting, and interbody fusion combined with posterior percutaneous pedicle screw fixation for lumbar spinal tuberculosis / 中国骨伤

OBJECTIVE@#To investigate the clinical efficacy and superiority of direct lateral interbody fusion combined with posterior percutaneous screw fixation in the treatment of lumbar tuberculosis.@*METHODS@#From June 2013 to August 2016, the clinical data of 83 patients with lumbar tuberculosis were retrospectively analyzed, including 55 males and 28 females, aged from 27 to 72 (49.5±13.5) years. These 83 patients were divided into two groups according to different operation methods, 35 cases in group A were treated with direct lateral interbody fusion combined with posterior percutaneous screw fixation;48 cases in group B were treated with anterior traditional extraperitoneal debridement combined with posterior internal fixation. After operation, regular quadruple antituberculosis drugs were continued for 18 months. The operation time, intraoperative blood loss, hospital stay, bone graft fusion time and complications were compared between the two groups. Visual analogue score (VAS) of lumbar pain, Oswestry Disability Index (ODI), sagittal Cobb angle, erythrocyte sedimentation rate (ESR) and C-reactive protein(CRP) values before and after operation were analyzed.@*RESULTS@#The operation was successfully completed in both groups, and the operation mode was not changed during operation. The operation time, intraoperative blood loss and hospital stay were (149.4±13.3) min, (354.3±69.0) ml, (9.4±1.6) d in group A and(116.8±10.0) min, (721.9±172.3) ml, (11.8±1.7) d in group B, respectively, with significant difference between the two groups (@*CONCLUSION@#The two kinds of operation can obtain satisfactory clinical effect. Direct lateral interbody fusion combined with posterior percutaneous screw fixation can reduce intraoperative blood loss and hospital stay, which is conducive to early rehabilitation of patients.

Clinicopathological features and prognosis of pleomorphic giant cell adenocarcinoma of the prostate / 中华泌尿外科杂志

Objective:To investigate the clinicopathological features and prognosis of pleomorphic giant cell adenocarcinoma (PGCA) of the prostate, and to strengthen the understanding of this rare variant.Methods:From January 2009 to December 2019, 383 pathological samples of prostate adenocarcinoma with Gleason scores of 8-10 were selected from the First Affiliated Hospital, Army Medical University. PGCA was screened by reviewing the histomorphology of hematoxylin and eosin stained sections. Then the expression of prostate specific markers and mismatch repair (MMR) proteins of PGCA were detected by immunohistochemistry (IHC), and microsatellite instability (MSI) status was detected through polymerase chain reaction (PCR)-capillary electrophoresis. Meanwhile, the clinicopathological characteristics, diagnosis, treatment and prognosis of PGCA were summarized and analyzed along with those reported in the literature.Results:Three patients with PGCA of the prostate were 68, 63 and 71 years old respectively, and case 1 had a history of transurethral resection of the prostate and oral bicalutamide 3 months before surgery. All 3 patients underwent radical prostatectomy and received endocrine therapy, radiotherapy and/or chemotherapy, and died at 18, 23, and 10 months after surgery, respectively. Histologically, both the usual prostate adenocarcinoma with Gleason score of 9-10 and the pleomorphic giant cell component with anaplastic characteristics were observed in 3 tumors, and the latter accounted for 90%, 10%, and 20%, respectively. Immunohistochemical staining showed that both components expressed epithelial markers (CK, CK8/18) and prostate-specific markers (NKX3.1, PSA, P504S) to varying degrees, and the expression of MMR proteins (MSH2, MSH6, MLH1 and PMS2) were not defective. MSI was not detected in the usual prostate adenocarcinoma and pleomorphic giant cell components obtained by microdissection in 3 cases. Combined with 10 cases reported in the literature, there were totally 13 cases of PGCA for reviewing. The patients were 45-81 years old, the average age was 66 years old, and the median age was 66 years old. During the follow-up period of 3-36 months, 7 cases relapsed/metastasized, 6 cases died within 23 months after diagnosis, and 4 of which died within 1 year.Conclusions:PGCA is a newly recognized rare variant of prostate adenocarcinoma. At present, all cases are accompanied with high-grade usual prostate adenocarcinoma with Gleason score of 9-10, but it is different from the latter in pathological morphology and clinical manifestations, by presenting high invasiveness and poor prognosis. PGCA is not sensitive to conventional endocrine therapy, radiotherapy or chemotherapy. Accurate diagnosis of PGCA is helpful to judge the prognosis of patients and guide the treatment.

One stage temporary atlantoaxial segmental fixation and reduction for Grauer type IIB dens fractures in teenagers / 中国骨伤

OBJECTIVE@#To assess the clinical results of one stage temporary atlantoaxial segmental fixation and reduction for Grauer type IIB dens fractures in teenagers.@*METHODS@#From February 2009 to April 2015, 19 teenagers with Grauer type IIB dens fractures not amenable to anteiror screw fixation were enrolled and treated using one stage temporary atlantoaxial segmental fixation and reduction without fusion. There were 14 males and 5 females, aged from 14 to 32 years with an average of (24.6±5.0 ) years. The internal fixation was removed after bone healing confirmed by CT scan. At the last follow-up (at least 1 year after internal fixation removal), dynamic CT was used to assess the atlantoaxial rotation activity. Visual analogue scale (VAS) was recorded before the first operation, before the second operation (removal of internal fixation) and at the last follow-up. Neck Disability Index(NDI) was used to evaluate the efficacy before the second operation (removal of internal fixation) and the last follow-up.@*RESULTS@#After operation, 2 patients developed the symptoms of occipital nerve stimulation such as numbness and pain in the occipitocervical region, and were treated with drugs such as dehydration and neurotrophic drugs, and the symptoms were relieved after 1 to 2 months. All the internal fixations were removed and all the patients were followed up more than 1 year, with time ranging from 18 to 25 months and an average of (21.47±2.41) months. The time of bone fusion after operation was 6 to 10 months with the mean of(8.21±1.27) months. Secondary surgical removal of internal fixation were performed immediately after fracture healing without internal fixation failure. The symptoms of neck pain improved significantly after operation, VAS score decreased from 6.74±0.65 before operation to 0.42±0.51 at the last follow-up after the second operation (removal of internal fixation), with statistically significant differences(<0.01). The NDI value decreased from (10.58±2.04)% before the second operation (removal of internal fixation) to (3.79±2.23)% at the last follow-up after the second operation (removal of internal fixation), with statistically significant difference(<0.01). At the last follow-up after the second operation (removal of internal fixation), dynamic CT showed that the unilateral rotation of the atlantoaxial spine reached (15.73±5.57)° to the left, (15.55±5.78)° to the right, and the overall rotation of the atlantoaxial spine was (31.28±10.71)°.@*CONCLUSIONS@#One stage temporary atlantoaxial segmental fixation and reduction for the treatment of Grauer type IIB dens fractures not amenable to anteiror screw fixation in teenagers can avoid the loss of atlantoaxial rotation function caused by atlantoaxial fusion, and to some extent retain the rotation activity of atlanto-axial joint.

Frictional resistance of archwires with various ligations in alignment phase / 中国组织工程研究

BACKGROUND: Clinical ligation of traditional brackets by stainless steel ligature wires has greater randomness among doctors, and previous studies have mainly concerned on brackets and wires. There are less studies on the differential frictional force due to tight or loose ligation, and moreover, how to define tight ligation or loose ligation is rarely reported. OBJECTIVE: To compare the frictional force of different archwires in multiple brackets with various ligations. METHODS: A plaster model with moderate crowded malocclusion was randomly selected, and copied by 3D digital scanning technologies and Objet30 OrthoDesk3D printing technologies. Then, the models were bonded with Damon Clear brackets, Damon 3MX brackets, 3M traditional MBT brackets and Habit monocrystal brackets. Frictional forces of different brackets on models combined with two types of low frictional NiTi archwires which were 0.30 mm (0.012 inch) and 0.36 mm (0.014 inch) were measured by microcomputer servo dynamometer. Traditional brackets were ligatured by tight ligation and loose ligation, respectively. RESULTS AND CONCLUSION: (1) When low-friction NiTi archwires of 0.30 mm were used, significant differences in the frictional force were found between different brackets with various ligations except for Damon Clear bracket versus 3M bracket with loose ligation as well as Damon Clear bracket versus Damon 3MX bracket (P ＜ 0.05). (2) When low-friction NiTi archwires of 0.36 mm were used, similarity in the frictional force was found between 3M bracket with loose ligation, Damon Clear bracket and Damon 3MX bracket, while there were no significant differences between 3M bracket with loose ligation and Damon 3MX, between 3M bracket with tight ligation and Habit bracket with loose ligation, as well as between Damon Clear bracket and 3M bracket with loose ligation (P ＞ 0.05). Comparing two kinds of archwires, the friction force showed no statistical difference between Damon Clear bracket and 3M bracket with loose and tight ligation (P ＞ 0.05), but a statistical difference was found between Damon 3MX bracket and Habit bracket (P ＜ 0.001), whatever two conventional brackets with tight ligation or loose ligation. To conclude, conventional MBT bracket with loose ligation can reduce frictional force as similar as self-ligating bracket in the alignment phase, and the reduction in the friction force is impacted by archwire size and bracket material.

Female robotic-assisted laparoscopic radical cystectomy with intracorporeal orthotopic W-shaped ileal neobladder / 中华泌尿外科杂志

Objective To report the technique and effect of robotic-assisted laparoscopic radical cystectomy (RARC) totally intracorporeal orthotopic ileal neobladder in female bladder cancer patient.Methods A consecutive series of 5 female patients with urothelial carcinoma of the bladder,who underwent RARC,bilateral pelvic lymph node dissection and intracorporeal orthotopic W-shaped ileal neobladder by an experienced robotic surgeon,were included in the retrospective study,from December 2017 to March 2018.Data were reviewed retrospectively.The patients aged 51-68 years old,with mean age 62.2 years,and BMI was 13.6-22.8 kg/m2,mean 18.1 kg/m2.The specimens were removed from the vagina.A 40 cm bowel segment with detubularisation was chosen for the W-shaped neobladder which was constructed manually.All patients resumed ambulation on postoperative 2 days and resumed oral feeding on postoperative day 3.The single-J stents and catheter were removed 12 days and 2 weeks after operation,respectively.Results All operation were performed successfully.The average operation time was 384.2 min (ranging 355-428 min) with mean estimated blood loss of 620 ml (ranging 150-1 800 ml).And the average number of dissected lymph nodes was 15 (ranging 10-20).The neobladder and urethra anastomotic leakage was observed in one patient who was healed after 2 weeks indwell catheter.Small bowel obstruction occurred in one case 3 weeks after operation and recovered after conservative treatment.The daytime urinary control was good,and 1 cases had urinary incontinence at night (follow up 4-7months,median:5.5months).No recurrence or metastasis was observed during routine follow-up.The imaging showed the good morphology of the upper urinary tract and the new bladder.Conclusions Our initial experience supports the feasibility and advantage of totally intracorporeal orthotopic W-shaped ileal neobladder following RARC in female patient with bladder cancer.It is worth to further verification in a large sample with longer follow-up.

VSITA, an Improved Approach of Target Amplification in the Identification of Viral Pathogens / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Unbiased next generation sequencing (NGS) is susceptible to interference from host or environmental sequences. Consequently, background depletion and virome enrichment techniques are usually needed for clinical samples where viral load is much lower than background sequences.</p><p><b>METHODS</b>A viral Sequence Independent Targeted Amplification (VSITA) approach using a set of non-ribosomal and virus-enriched octamers (V8) was developed and compared with traditionally used random hexamers (N6). Forty-five archived clinical samples of different types were used in parallel to compare the V8 and N6 enrichment performance of viral sequences and removal performance of ribosomal sequences in the step of reverse transcription followed by quantitative PCR (qPCR). Ten sera samples from patients with fever of unknown origin and 10 feces samples from patients with diarrhea of unknown origin were used in comparison of V8 and N6 enrichment performance following NGS analysis.</p><p><b>RESULTS</b>A minimum 30 hexamers matching to viral reference sequences (sense and antisense) were selected from a dataset of random 4,096 (46) hexamers (N6). Two random nucleotides were added to the 5' end of the selected hexamers, and 480 (30 × 42) octamers (V8) were obtained. In general, VSITA approach showed higher enrichment of virus-targeted cDNA and enhanced ability to remove unwanted ribosomal sequences in the majorities of 45 predefined clinical samples. Moreover, VSITA combined with NGS enabled to detect not only more viruses but also achieve more viral reads hit and higher viral genome coverage in 20 clinical samples with diarrhea or fever of unknown origin.</p><p><b>CONCLUSION</b>The VSITA approach designed in this study is demonstrated to possess higher sensitivity and broader genome coverage than traditionally used random hexamers in the NGS-based identification of viral pathogens directly from clinical samples.</p>

Factors Associated with Trait Anger Level of Juvenile Offenders in Hubei Province: A Binary Logistic Regression Analysis / 华中科技大学学报（医学）（英德文版）

The risk factors of high trait anger of juvenile offenders were explored through question naire study in a youth correctional facility of Hubei province,China.A total of 1090 juvenile offenders in Hubei province were investigated by self-compiled social-demographic questionnaire,Childhood Trauma Questionnaire (CTQ),and State-Trait Anger Expression Inventory-Ⅱ (STAXI-Ⅱ).The risk factors were analyzed by chi-square tests,correlation analysis,and binary logistic regression analysis with SPSS 19.0.A total of 1082 copies of valid questionnaires were collected.High trait anger group (n=316) was defined as those who scored in the upper 27th percentile of STAXI-Ⅱ trait anger scale (TAS),and the rest were defined as low trait anger group (n=766).The risk factors associated with high level of trait anger included:childhood emotional abuse,childhood sexual abuse,step family,frequent drug abuse,and frequent internet using (P＜0.05 or P＜0.01).Birth sequence,number of sibling,ranking in the family,identity of the main care-taker,the education level of care-taker,educational style of care-taker,family income,relationship between parents,social atmosphere of local area,frequent drinking,and frequent smoking did not predict to high level of trait anger (P＞0.05).It was suggested that traumatic experience in childhood and unhealthy life style may significantly increase the level of trait anger in adulthood.The risk factors of high trait anger and their effects should be taken into consideration seriously.

The whole-cell immobilization of D-hydantoinase-engineered Escherichia coli for D-CpHPG biosynthesis

Background: D-Hydroxyphenylglycine is considered to be an important chiral molecular building-block of antibiotic reagents such as pesticides, and β-lactam antibiotics. The process of its production is catalyzed by D-hydantoinase and D-carbamoylase in a two-step enzyme reaction. How to enhance the catalytic potential of the two enzymes is valuable for industrial application. In this investigation, an Escherichia coli strain genetically engineered with D-hydantoinase was immobilized by calcium alginate with certain adjuncts to evaluate the optimal condition for the biosynthesis of D-carbamoyl-p-hydroxyphenylglycine (D-CpHPG), the compound further be converted to D-hydroxyphenylglycine (D-HPG) by carbamoylase. Results: The optimal medium to produce D-CpHPG by whole-cell immobilization was a modified Luria-Bertani (LB) added with 3.0% (W/V) alginate, 1.5% (W/V) diatomite, 0.05% (W/V) CaCl2 and 1.00 mM MnCl2.The optimized diameter of immobilized beads for the whole-cell biosynthesis here was 2.60 mm. The maximized production rates of D-CpHPG were up to 76%, and the immobilized beads could be reused for 12 batches. Conclusions: This investigation not only provides an effective procedure for biological production of D-CpHPG, but gives an insight into the whole-cell immobilization technology.

Involvement of α5 integrin in survivin-mediated osteosarcoma metastasis

OBJECTIVE@#To investigate the role of survivin in osteosarcoma metastasis.@*METHODS@#Small interfering RNA (siRNA) was used to knockdown the expression of survivin and α5 integrin in the human osteosarcoma cell line MG63. Western blotting and immunostaining methods were used to assessed the effect of survivin knockdown on the expression of α5 integrin through flow cytometry and fluorescence microscopy detection. Meanwhile, the invasion and migration of transfected cells in Transwell and wound healing assays were probed, and the growth situation of these cells transplanted into nude mice was monitored.@*RESULTS@#Knockdown of survivin expression could inhibit the invasion and migration of osteosarcoma MG64 cells in vitro and the expression of α5 integrin on osteosarcoma MG64 cell surface, suggesting that survivin can inhibit the invasion and migration of osteosarcoma cells through downregulation of α5 integrin. Anti-α5 integrin antibody could also markedly decrease the capability of invasion and migration of osteosarcoma MG64 cells. Additionally, knockdown of survivin expression could slow the growth of osteosarcoma MG63 cells transplanted into nude mice.@*CONCLUSIONS@#Survivin-directed anti-tumor strategies might be an effective method in the treatment of osteosarcoma.

Marsdeniae tenacissimae extract (MTE) suppresses cell proliferation by attenuating VEGF/VEGFR2 interactions and promotes apoptosis through regulating PKC pathway in human umbilical vein endothelial cells / 中国天然药物

Marsdeniae tenacissimae extract (MTE), commonly known as Xiao-Ai-Ping in China, is a traditional Chinese herb medicine capable of inhibiting proliferation and metastasis and boosting apoptosis in various cancer cells. However, little is known about the contribution of MTE towards tumor angiogenesis and the underlying mechanism. The present study aimed to evaluate the effects of MTE on the proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs) and the molecular mechanism. 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium, inner salt (MTS) and PI-stained flow cytometry assays revealed that MTE dose-dependently reduced the proliferation of HUVECs by arresting cell cycle at S phase (P < 0.05). Annexin V-FITC/PI-stained flow cytometry confirmed that MTE (160 μL·L) enhanced the apoptosis of HUVECs significantly (P < 0.001). Real-time quantitative RT-PCR and Western blot analyses showed an increase in Bax expression and a sharply decline in Bcl-2 expression; caspase-3 was activated simultaneously in a dose-dependent manner (P < 0.05). Further study observed the dose-dependent down-regulation of vascular endothelial growth factor (VEGF) receptor-2 (VEGFR-2), P2Y6 receptor (P2Y6R), and chemokine (C-C motif) ligand 2 (CCL-2), along with the activation of PKC Δ and up-regulation of p53 in a dose-dependent manner in MTE-treated selected cells (P < 0.05). Collectively, the results from the present study suggested that MTE suppressed the proliferation by attenuating CCL-2-mediated VEGF/VEGFR2 interactions and promoted the apoptosis through PKCΔ-induced p53-dependent mitochondrial pathway in HUVECs, supporting that MTE may be developed as a potent anti-cancer medicine.

Involvement of α5 integrin in survivin-mediated osteosarcoma metastasis

Objective To investigate the role of survivin in osteosarcoma metastasis. Methods Small interfering RNA (siRNA) was used to knockdown the expression of survivin and α5 integrin in the human osteosarcoma cell line MG63. Western blotting and immunostaining methods were used to assessed the effect of survivin knockdown on the expression of α5 integrin through flow cytometry and fluorescence microscopy detection. Meanwhile, the invasion and migration of transfected cells in Transwell and wound healing assays were probed, and the growth situation of these cells transplanted into nude mice was monitored. Results Knockdown of survivin expression could inhibit the invasion and migration of osteosarcoma MG64 cells in vitro and the expression of α5 integrin on osteosarcoma MG64 cell surface, suggesting that survivin can inhibit the invasion and migration of osteosarcoma cells through downregulation of α5 integrin. Anti-α5 integrin antibody could also markedly decrease the capability of invasion and migration of osteosarcoma MG64 cells. Additionally, knockdown of survivin expression could slow the growth of osteosarcoma MG63 cells transplanted into nude mice. Conclusions Survivin-directed anti-tumor strategies might be an effective method in the treatment of osteosarcoma.

Multipore-light brown fixation prevents PICC catheter from prolapsing / 现代临床护理

Objective To explore the effect of multipore-light brown fixation on preventing peripherally inserted central catheter from prolapsing. Methods Randomizedly 82 patients with PICC were equally divided into treatment group and control group. Multipore-Light Brown fixation was used in the treatment group to stabilize catheter , while the routine care was applied in the control group for stabilization . The two groups were compared in terms of prolapse rate and degree of the PICC . Result The prolapse rate and the prolapse degree of the treatment group were significantly lower than that of the control group ( P < 0 . 01 ) . Conclusions The multipore-light brown fixation is more effective in the stabilization of PICC. It can effectively prevent the PICC from prolapsing.

Direct measurement of blood nickel using compound matrix modifier-graphite furnace atomic absorption spectrometry / 中国职业医学

OBJECTIVE: To establish a methodology to dilute blood sample with compound matrix modifier,and to determine the blood level of nickel using improved graphite furnace atomic absorption spectrometry( GFAAS). METHODS: The blood samples( 100. 00 μL) were diluted 5 times in 0. 12 g / L mass concentration of palladium chloride combined with 1. 00 %(V /V) Triton X-100 and 0. 10%(V /V) nitric acid,and the blood level of nickel was detected by GFAAS. RESULTS: The good linearity range of nickel mass concentration in blood was 0. 30-25. 00 μg / L,the correlation coefficient was 0. 999 7,the detection limit was 0. 30 μg / L,the minimum detectable mass concentration was 1. 50 μg / L,the recovery rate was99. 00 %-102. 75 %. The within-run and between-run relative standard deviations were 1. 43 %-3. 70 % and 2. 20 %-5. 03 %,respectively. Blood samples can be stored for at least 7 days at 4 ℃. CONCLUSION: This method is simple for operation,with high accuracy and precision,which is suitable for determination of nickel in blood in normal individuals and workers with occupational exposed to nickel.

Comparative analysis of biological characteristics of adult mesenchymal stem cells with different tissue origins

Objective: To invest the differences among mesenchymal stem cells (MSCs) derived from different tissues and their impacts on clinical applications. Methods: In this study, MSCs were isolated from adipose tissue (AD), umbilical cord tissue (UC), and menstrual blood (Men) and compared their biological characteristics in terms of proliferation capacity, passage capacity, colony formation, and surface markers were compared. Results: The stem cells (SCs) obtained from different sources were all characterized as MSCs, but demonstrated some differences. Umbilical cord-derived MSCs (UCMSCs) were able to overcome density inhibition. The proliferation rate decreased in the order UCMSCs > MenSCs > ADSCs, while the colony-forming ability decreased in the order MenSCs > ADSCs > UCMSCs. Based on gene-expression data for MSCs from different sources within the same donor, 768 MenSC genes were found that were specifically upregulated or downregulated compared with bone marrow-derived MSCs and UCMSCs, most of which were involved in cell function-related pathways. In addition, MenSCs appeared to be superior in terms of immune inflammation, stress response, and neural differentiation potentials, but weaker in terms of osteogenic and chondrogenic differentiation capacities, compared with UCMSCs and bone marrow-derived MSCs. Conclusions: MenSCs have higher extraction efficiency, colony-forming ability, and long time passage capacity. Although the proliferation capacity is inferior to UCMSCs.

Comparative analysis of biological characteristics of adult mesenchymal stem cells with different tissue origins

OBJECTIVE@#To invest the differences among mesenchymal stem cells (MSCs) derived from different tissues and their impacts on clinical applications.@*METHODS@#In this study, MSCs were isolated from adipose tissue (AD), umbilical cord tissue (UC), and menstrual blood (Men) and compared their biological characteristics in terms of proliferation capacity, passage capacity, colony formation, and surface markers were compared.@*RESULTS@#The stem cells (SCs) obtained from different sources were all characterized as MSCs, but demonstrated some differences. Umbilical cord-derived MSCs (UCMSCs) were able to overcome density inhibition. The proliferation rate decreased in the order UCMSCs > MenSCs > ADSCs, while the colony-forming ability decreased in the order MenSCs > ADSCs > UCMSCs. Based on gene-expression data for MSCs from different sources within the same donor, 768 MenSC genes were found that were specifically upregulated or downregulated compared with bone marrow-derived MSCs and UCMSCs, most of which were involved in cell function-related pathways. In addition, MenSCs appeared to be superior in terms of immune inflammation, stress response, and neural differentiation potentials, but weaker in terms of osteogenic and chondrogenic differentiation capacities, compared with UCMSCs and bone marrow-derived MSCs.@*CONCLUSIONS@#MenSCs have higher extraction efficiency, colony-forming ability, and long time passage capacity. Although the proliferation capacity is inferior to UCMSCs.

Impact of sacral nerve root resection on the erectile and ejaculatory function of the sacral tumor patient / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the erectile and ejaculatory function of sacral tumor patients after sacral nerve root resection and investigate the relationship of erectile and ejaculatory dysfunction (EED) with the level of sacral nerve injury.</p><p><b>METHODS</b>This retrospective study included 47 male patients aged 16 to 63 (32.6 +/- 6.8) years treated by sacral tumor resection between January 2008 and August 2013. According to the levels of the sacral nerve roots spared in surgery, the patients were divided into four groups: bilateral S1-S3 (n=16), unilateral S1-S3 (n=21), unilateral S1-S2 (n=6), and unilateral S1 (n=4). The patients were followed up for 12 to 41 (27.2 +/- 10.9) months by questionnaire investigation, clinic review, and telephone calls about their erectile and ejaculatory function at 3, 6 and 12 months after surgery and in August 2013.</p><p><b>RESULTS</b>In the bilateral S1-S3 group, the incidence rates of EED were 31.25% (5/16), 25% (4/16), and 12.5% (2/16) at 3, 6, and 12 months respectively after surgery, with recovery of erectile and ejaculatory function in August 2013. The incidence rates of EED in the unilateral S1-S3 group were 85.71% (18/21), 71.43% (15/21), 52.38% (11/21), and 42.86% (9/21) at 3, 6 and 12 months and in August 2013, respectively; those in the unilateral S1-S2 group were 100% (6/6), 83.33% (5/6), 83.33% (5/6), and 66.67% (4/6) at the four time points; and those in the unilateral S1 group were all 100% (4/4). No statistically significant differences were found in the incidence rate of EED among the patients of different ages or tumor types (P > 0.05).</p><p><b>CONCLUSION</b>The incidence of postoperative EED in male patients treated by sacral tumor resection is closely related to the mode of operation. Sparing the S3 nerve root at least unilaterally in sacral tumor resection is essential for protecting the erectile and ejaculatory function of the patient.</p>

miRNA-101 inhibits the expression of the enhancer of zeste homolog 2 in androgen-independent prostate cancer LNCaP cell line / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.</p><p><b>METHODS</b>We divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.</p><p><b>RESULTS</b>Green fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.</p><p><b>CONCLUSION</b>miRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.</p>