Biallelic mutations in spermatogenesis and centriole-associated 1 like (SPATC1L) cause acephalic spermatozoa syndrome and male infertility / 亚洲男科学杂志(英文版)

Acephalic spermatozoa syndrome is a rare type of teratozoospermia that severely impairs the reproductive ability of male patients, and genetic defects have been recognized as the main cause of acephalic spermatozoa syndrome. Spermatogenesis and centriole-associated 1 like (SPATC1L) is indispensable for maintaining the integrity of sperm head-to-tail connections in mice, but its roles in human sperm and early embryonic development remain largely unknown. Herein, we conducted whole-exome sequencing (WES) of 22 infertile men with acephalic spermatozoa syndrome. An in silico analysis of the candidate variants was conducted, and WES data analysis was performed using another cohort consisting of 34 patients with acephalic spermatozoa syndrome and 25 control subjects with proven fertility. We identified biallelic mutations in SPATC1L (c.910C>T:p.Arg304Cys and c.994G>T:p.Glu332X) from a patient whose sperm displayed complete acephalia. Both SPATC1L variants are rare and deleterious. SPATC1L is mainly expressed at the head-tail junction of elongating spermatids. Plasmids containing pathogenic variants decreased the level of SPATC1L in vitro. Moreover, none of the patient's four attempts at intracytoplasmic sperm injection (ICSI) resulted in a transplantable embryo, which suggests that SPATC1L defects might affect early embryonic development. In conclusion, this study provides the first identification of SPATC1L as a novel gene for human acephalic spermatozoa syndrome. Furthermore, WES might be applied for patients with acephalic spermatozoa syndrome who exhibit reiterative ICSI failures.

A 14-year multi-institutional collaborative study of Chinese pelvic floor surgical procedures related to pelvic organ prolapse / 中华医学杂志(英文版)

BACKGROUND@#It has been a global trend that increasing complications related to pelvic floor surgeries have been reported over time. The current study aimed to outline the development of Chinese pelvic floor surgeries related to pelvic organ prolapse (POP) over the past 14 years and investigate the potential influence of enhanced monitoring conducted by the Chinese Association of Urogynecology since 2011.@*METHODS@#A total of 44,594 women with POP who underwent pelvic floor surgeries between October 1, 2004 and September 30, 2018 were included from 22 tertiary academic medical centers. The data were reported voluntarily and obtained from a database. We compared the proportion of each procedure in the 7 years before and 7 years after September 30, 2011. The data were analyzed by performing Z test (one-sided).@*RESULTS@#The number of different procedures during October 1, 2011-September 30, 2018 was more than twice that during October 1, 2004-September 30, 2011. Regarding pelvic floor surgeries related to POP, the rate of synthetic mesh procedures increased from 38.1% (5298/13,906) during October 1, 2004-September 30, 2011 to 46.0% (14,107/30,688) during October 1, 2011-September 30, 2018, whereas the rate of non-mesh procedures decreased from 61.9% (8608/13,906) to 54.0% (16,581/30,688) (Z = 15.53, P < 0.001). Regarding synthetic mesh surgeries related to POP, the rates of transvaginal placement of surgical mesh (TVM) procedures decreased from 94.1% (4983/5298) to 82.2% (11,603/14,107) (Z = 20.79, P < 0.001), but the rate of laparoscopic sacrocolpopexy (LSC) procedures increased from 5.9% (315/5298) to 17.8% (2504/14,107).@*CONCLUSIONS@#The rate of synthetic mesh procedures increased while that of non-mesh procedures decreased significantly. The rate of TVM procedures decreased while the rate of LSC procedures increased significantly.@*TRIAL REGISTRATION NUMBER@#NCT03620565, https://register.clinicaltrials.gov.

Study on the structure of eye anterior segment after laser treatment in ROP with Sirius anterior segment analysis system / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To observe the changes of the development of the anterior segment in children after the treatment of laser photocoagulation(LP)for retinopathy of prematurity(ROP)by corneal topographic. <p>METHODS: In this retrospective case study, 25 children(50 eyes)as ROP group and 23 children(46 eyes)born at term as control group had participated in the study. The best corrected visual acuity(BCVA)was examined in both groups, and converted to LogMAR vision when statistical analysis was performed. The observation indicators of the Sirius anterior segment analysis system in the study were as follows: the horizontal iris diameter(HVID), corneal radius of the thinnest point, the thinnest point of the cornea, the maximum curvature of the cornea, the maximum radius of cornea curvature, central corneal thickness(CCT), corneal volume(CV), anterior chamber depth(ACD), anterior chamber volume and the anterior chamber angle.<p>RESULTS: The HVID, the thinnest point of the cornea, ACD, and the anterior chamber volume in ROP group were smaller than that in the control group(all <i>P</i><0.05). The CV and the anterior chamber angle in ROP group were smaller than that in the control group, but there was no significant difference between the two groups(all <i>P</i>>0.05). The BCVA was significantly better in the control group than that in the ROP group(0.07±0.10 <i>vs</i> 0.24±0.25, <i>P</i><0.05). <p>CONCLUSION: The development of eye anterior ganglion tissues of the ROP group had changed with steeper cornea, shallower anterior chamber, smaller angle of the anterior chamber and poor BCVA. All of the changes above might make it easier for the development of refractive errors and glaucoma.

Discussion on the effect of comprehensive nursing in endoscopic treatment of esophageal and gastric varicose bleeding / 中国实用护理杂志

Objective:To explore and analyze the effect of comprehensive nursing care in endoscopic treatment of esophageal and gastric varicose bleeding.Methods:A retrospective analysis of 60 cases of patients with esophageal and gastric varicose hemorrhage admitted to the hospital was conducted. The patients were randomly divided into 30 cases of control group and 30 cases of observation group. Routine nursing and comprehensive nursing were adopted respectively to evaluate different nursing effects.Results:Intraoperative and postoperative heart rate of the observation group (91.0 ± 4.0) times/min, (89.2 ± 4.2) times/min, which were lower than (98.0 ± 4.2) times/min, (94.7 ± 4.5) times/min of the control group. There was statistically significant difference ( t values were 6.610, 4.894, P < 0.05). The bleeding prevention effectiveness rate of the observation group (93.33%, 28/30) was higher than the control group (73.33%, 22/30), The difference was statistically significant ( χ 2 value was 4.32, P < 0.05). The total nursing satisfaction rate (100.00%, 30/30) in the observation group was higher than that in the control group (86.70%, 26/30), the difference was statistically significant ( χ 2 value was 4.29, P< 0.05). Quality of life, disease cognition, compliance behavior, proper rest, self-protection, General Quality of life Inventory (GQOLI) score in the observation group were (33.5 ± 3.5), (21.5 ± 3.7), (32.5 ± 2.8), (21.5 ± 3.3), (22.0±4.2), (130.2 ± 17.4) points, which were higher than those in the control group (28.2 ± 3.2), (17.5 ± 2.5), (28.5 ± 3.0), (17.2 ± 3.0), (18.0 ± 3.0), and (110.2 ± 15.2) points. The differences were statistically significant ( t values were 4.245-6.121, P<0.05). Conclusions:Endoscopic treatment of esophageal and gastric varicose bleeding patients combined with comprehensive nursing intervention, can effectively prevent the occurrence of rebleeding, improve the quality of life of patients. The application effect is significant.

Discussion on the effect of comprehensive nursing in endoscopic treatment of esophageal and gastric varicose bleeding / 中国实用护理杂志

Objective@#To explore and analyze the effect of comprehensive nursing care in endoscopic treatment of esophageal and gastric varicose bleeding.@*Methods@#A retrospective analysis of 60 cases of patients with esophageal and gastric varicose hemorrhage admitted to the hospital was conducted. The patients were randomly divided into 30 cases of control group and 30 cases of observation group. Routine nursing and comprehensive nursing were adopted respectively to evaluate different nursing effects.@*Results@#Intraoperative and postoperative heart rate of the observation group (91.0 ± 4.0) times/min, (89.2 ± 4.2) times/min, which were lower than (98.0 ± 4.2) times/min, (94.7 ± 4.5) times/min of the control group. There was statistically significant difference (t values were 6.610, 4.894, P < 0.05). The bleeding prevention effectiveness rate of the observation group (93.33%, 28/30) was higher than the control group (73.33%, 22/30), The difference was statistically significant (χ2 value was 4.32, P < 0.05). The total nursing satisfaction rate (100.00%, 30/30) in the observation group was higher than that in the control group (86.70%, 26/30), the difference was statistically significant (χ2 value was 4.29, P< 0.05). Quality of life, disease cognition, compliance behavior, proper rest, self-protection, General Quality of life Inventory (GQOLI) score in the observation group were (33.5 ± 3.5), (21.5 ± 3.7), (32.5 ± 2.8), (21.5 ± 3.3), (22.0±4.2), (130.2 ± 17.4) points, which were higher than those in the control group (28.2 ± 3.2), (17.5 ± 2.5), (28.5 ± 3.0), (17.2 ± 3.0), (18.0 ± 3.0), and (110.2 ± 15.2) points. The differences were statistically significant (t values were 4.245-6.121, P<0.05).@*Conclusions@#Endoscopic treatment of esophageal and gastric varicose bleeding patients combined with comprehensive nursing intervention, can effectively prevent the occurrence of rebleeding, improve the quality of life of patients. The application effect is significant.

Comparison of clinical efficacy of complete and incomplete radical debridement for spinal tuberculosis: a Meta-analysis / 中国骨伤

<p><b>OBJECTIVE</b>To compare clinical efficacy of complete and incomplete radical debridement for spinal tuberculosis by Meta-analysis.</p><p><b>METHODS</b>The literatures of RCT or non-RCT with complete and incomplete radical debridement for spinal tuberculosis from Medline, EMBASE, Cochrane Library, Web of Science, CBM, CNKI and Wanfang were searched from the time of creating database to July, 2017. Two independent reviewers identified eligible studies, extracted data and evaluated risk of bias of included studies. Meta analysis were performed by Revman 5.3 and GRADE system were used to grade evidence. Recurrence rate, adverse effects, healing time, chemotherapy duration, spinal deformity by correction angle, bone fusion time in interface of intervertebral, erythrocyte sedimentation rate and C-reaction protein were compared between two groups.</p><p><b>RESULTS</b>Totally 9 literatures were chosen, including 5 RCT and 4 non-RCT with 1 302 patients. Compared with incomplete radical debridement, complete radical debridement had lower recurrence rate [=0.14, 95%CI(0.08, 0.22), <0.000 01], lower rate of adverse effects[=0.18, 95%CI(0.12, 0.27), <0.000 01], shorter healing time[MD=-4.80, 95%CI(-5.14, -4.45), <0.000 01]and chemotherapy duration [MD=-5.25, 95%CI(-5.64, -4.86), <0.000 01], larger spinal deformity by correction angle[MD=4.88, 95%CI(3.55, 6.27), <0.000 01], smaller erythrocyte sedimentation rate[MD=-8.74, 95%CI(-11.99, -5.49), <0.000 01] and C-reaction protein [MD=-4.75, 95%CI(-8.61, -0.88), =0.02] . However, there was no difference on bone fusion time in interface of intervertebral between two groups[MD=-0.19, 95%CI(-0.50, 0.12), =0.23].</p><p><b>CONCLUSIONS</b>Compared with incomplete radical debridement, complete radical debridement has advantages of lower incidence of recurrence, lower rate of adverse reaction, shorten healing time and chemotherapy time, recovered faster. Techniques are selected according to indication of patients individual, complete radical debridement is recommended at the same indications.</p>

Diffusion tensor imaging study of white matter fiber in patients with schizophrenia episode in early adulthood / 新乡医学院学报

Objective To explore the structure changes of white matter of the patients with schizophrenia episode in early adulthood by diffusion tensor imaging(DTI) and in order to provide a structural neuroimaging basis for understanding the pathology of schizophrenia.Methods Twenty-six patients with schizophrenia episode in early adulthood in the Second Affiliated Hospital of Xinxiang Medical University from July 2012 to March 2014 were selected as study group and twenty-eight healthy subjects whose age,sex,education were matched with study group were selected as control group.All subjects received structural magnetic resonance imaging and DTI scans.The fractional anisotropy (FA) and the mean diffusivity (MD) of the white matter of the same encephalic region were compared between the two groups by voxel-based analyses.Results The FA values of the right anterior cingulate gyrusthe,genu of corpus callosum,the right limb of internal capsule,the bilateral external capsule,the bilateral posterior of coronal radiate,the right anterior coronal radiate of patients in the study group were significantly lower than those in the control group (P ＜ 0.05).The MD values of the bilateral limbs of internal capsule,the right cingulate gyrus,left superior longitudinal tract,corpus callosum and right anterior coronal radiate of patients in the study group were significantly higher than those in the control group (P ＜ 0.05).Conclusion Schizophrenia patients who episode in early adulthood exist widespread microstructural damage of white matter.These changes may be related to the pathological change of schizophrenia.

Clinical comparison of percutaneous transluminal angioplasty and surgical resection for Cimino-Brescia arteriovenous fistula stenosis in hemodialysis patients / 医学研究生学报

Objeetive As to the high incidence of arteriovenous fistula(AVF) stenosis,surgical operation will result in the exhaustion of vascular resources in patients,while percutaneous transluminal angioplasty(PTA) can maintain vascular resources for ostomy.However,there is still no clear definition between the choices of PTA and surgical resection.The aim of this study was to compare the efficacy of PTA and surgical resection followed by reconstruction for the treatment of arteriovenous fistula stenosis in order to find appropriate treatment.Methods Retrospective analysis had been done on 46 hemodialysis patients with arteriovenous fistula stenosis in Nanjing BenQ hospital from January 2015 to March 2017,which included 22 cases treated with PTA (PTA group) and 24 cases treated with surgical operation (operation group).Comparison was made in general clinical situation,patency rate at six months after surgery,over patency time and adverse reactions to surgery between the two groups.Results The number of stenoses in PTA group was bigger than that in operation group and the difference was of statistic significance (2.78±1.43 vs 1.67±0.71,P＜0.05).There was no significant difference in patency rate between the two groups (P =0.828).There were 57 venous stenoses in PTA group,among which 12 stenoses were anastomotic (21.05％) with 79.3％ average stenosis degree and 43 stenoses were at venous outflow tract of fistula (75.44％) with 84.26 average stenosis degree.In PTA group,3 patients had hematoma brachial puncture position and recovered by self-absorption without special treatment.In operation group,1 patient had mild blood oozing and recovered after treatment;4 patients recovered gradually from mild swelling on the back of the hand of the operation side.No difference was found in adverse reactions between two groups (P＞0.05).Conclusion PTA treatment is preferred for multiple stenoses(n ≥ 3),which ensures better preservation of vascular resources at a comparable patency rate.

Risk Factors for Mesh Exposure after Transvaginal Mesh Surgery / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Mesh exposure after surgery continues to be a clinical challenge for urogynecological surgeons. The purpose of this study was to explore the risk factors for polypropylene (PP) mesh exposure after transvaginal mesh (TVM) surgery.</p><p><b>METHODS</b>This study included 195 patients with advanced pelvic organ prolapse (POP), who underwent TVM from January 2004 to December 2012 at the First Affiliated Hospital of Chinese PLA General Hospital. Clinical data were evaluated including patient's demography, TVM type, concomitant procedures, operation time, blood loss, postoperative morbidity, and mesh exposure. Mesh exposure was identified through postoperative vaginal examination. Statistical analysis was performed to identify risk factors for mesh exposure.</p><p><b>RESULTS</b>Two-hundred and nine transvaginal PP meshes were placed, including 194 in the anterior wall and 15 in the posterior wall. Concomitant tension-free vaginal tape was performed in 61 cases. The mean follow-up time was 35.1 ± 23.6 months. PP mesh exposure was identified in 32 cases (16.4%), with 31 in the anterior wall and 1 in the posterior wall. Significant difference was found in operating time and concomitant procedures between exposed and nonexposed groups (F = 7.443, P = 0.007; F = 4.307, P = 0.039, respectively). Binary logistic regression revealed that the number of concomitant procedures and operation time were risk factors for mesh exposure (P = 0.001, P = 0.043).</p><p><b>CONCLUSION</b>Concomitant procedures and increased operating time increase the risk for postoperative mesh exposure in patients undergoing TVM surgery for POP.</p>

Correlation of the inner diameter parameters of the spermatic vein in different positions and states of the varicocele patient with the results of seminal examination / 中华男科学杂志

<p><b>Objective</b>To study the correlation of the inner diameter parameters of the spermatic vein in different positions and states of the varicocele (VC) patient with the results of seminal examination.</p><p><b>METHODS</b>A total of 149 VC patients underwent ultrasonography, routine semen examination, and sperm morphological analysis. The parameters obtained from ultrasonography included the bilateral testis volume in a supine position, the largest spermatic vein diameter in a supine position at rest (DSR), the largest spermatic vein diameter in a supine position following Valsalva manoeuvre (DSV), the largest spermatic vein diameter in an upright position at rest (DUR), and the largest spermatic vein diameter in an upright position following Valsalva manoeuvre (DUV). Then we calculated the parameters △DS=DSV－DSR, △DU=DUV－DUR, △DR=DUR－DSR, and △DV=DUV－DSV and analyzed the correlation of the above parameters with the results of semen examination using the ROC curve.</p><p><b>RESULTS</b>Based on the results of semen examination, 119 (79.87%) of the patients were allocated to the abnormal group and the other 30 (20.13%) to the normal group. Statistically significant differences were observed between the two groups in △DU (P=0.007), △DR (P=0.0001), and △DV (P=0.04), but not in DSR (P=0.35), DSV (P=0.34), DUR (P=0.06), DUV (P=0.12), and △DS (P=0.64), nor in the volume of the testis affected (P=0.323). The area under the ROC curve was 0.55 for DSR, 0.57 for DSV, 0.64 for DUR, 0.62 for DUV, 0.49 for △DS, 0.28 for △DU, 0.86 for △DR, and 0.69 for △DV. The corresponding cutoff values were 2.25, 2.51, 2.48, 2.63, 0.30, 0.23, 0.25, and 0.20, the corresponding sensitivities of semen detection were 50.42%, 65.55%, 60.50%, 60.50%, 49.90%, 29.41%, 79.83%, and 65.55%, and the corresponding specificities were 56.67%, 63.33%, 63.33%, 63.33%, 56.67%, 33.33%, 80%, and 63.33%, respectively.</p><p><b>CONCLUSIONS</b>The difference between the largest spermatic vein diameters in supine and upright positions at rest provides a high diagnostic accuracy for semen detection and helps to predict abnormality in seminal examination for VC patients.</p>

Prognostic factors for hemophagocytic lymphohistiocytosis in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the main factors influencing prognosis of hemophagocytic lymphohistiocytosis (HLH) in children by summarizing the clinical features of HLH and investigating the relationship between relevant factors and prognosis.</p><p><b>METHODS</b>The medical data of 63 children with HLH were retrospectively reviewed. Kaplan-Meier method was employed to draw survival curves. Factors influencing prognosis were assessed with Cox univariate analysis, and Cox multivariate analysis was done on statistically significant factors.</p><p><b>RESULTS</b>The 3-year and 5-year survival rates were both 62.9%. The survival rate decreased from 98.4% at 1 day after definite diagnosis to 73.2% at 4 months. Univariate analysis demonstrated only one factor, which was that the condition of platelet recovery after treatment of 2 to 3 weeks was significantly related to prognosis (P=0.002). In children receiving etoposide therapy, temperature recovery after one day of treatment was significantly related to prognosis (P=0.016).</p><p><b>CONCLUSIONS</b>Children with HLH have a satisfactory prognosis, but the survival rate reduces rapidly in the first 4 months after definite diagnosis. Platelet recovery after treatment of 2 to 3 weeks and temperature recovery after one day of treatment are factors influencing prognosis of HLH in children.</p>

Gene expression profile changes caused by the dysfunction of Mer during retinal pigment epithelium phagocytosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Studies indicated that Mer might be the main contributor to the specific internalization of photoreceptor outer segments (POS) in retinal pigment epithelium (RPE). It is very important to understand the mechanism of POS phagocytosis under the pathway of Mer and its ligands. The objective of this study was to identify changes in gene expression profiles caused by Mer gene knockout (Mer-/-) during phagocytosis of POS in RPE.</p><p><b>METHODS</b>RPE from both Mer-/- and wild-type (WT) mice were isolated and cultured to the 3rd passage. POS were subjected to culture medium with 20 nmol/L Gas6 and protein S to activate specific mer-mediated phagocytosis. RPE phagocytosis was evaluated by phagocytosis assays and differential gene expression identified by microarray at 3 and 12 hours; the 0-hour time point served as the control. Three independent samples for each Mer-/- or WT RPE were subjected to the same protocol of microarray. Five genes were confirmed by real-time quantitative PCR (QPCR).</p><p><b>RESULTS</b>The Mer-/- RPE had less internalized POS than WT RPE after both 3 and 12 hours in phagocytosis assay. Compared to WT RPE and the 0-hour control, 38 and 45 different known genes were increased and 68 and 59 known genes were decreased in Mer-/- RPE after 3 and 12 hours, respectively. Abnormal POS phagocytosis in Mer-/- RPE was associated with significant gene expression changes in, for example, signal transduction (WNT, MAPK), phagocytosis (Vav3, Hsd11b1), cytoskeleton components (Myo7a), and metabolism, in a time-specific manner. QPCR results showed Vav3, Hsd11b1, Myo7a, Rtn2 and Itga8 in those independent samples were consistent with microarray.</p><p><b>CONCLUSION</b>Gene expression profiles modulated in a time-specific manner in Mer-/- RPE indicate a possible internalization mechanism for abnormal POS phagocytosis, which gives insight into the mechanism of retinitis pigmentosa caused by the mutation of MerTK in humans.</p>

Segmental meshing of brain tissues and simulation of soft tissue deformation / 南方医科大学学报

Virtual surgery provides important assistance in preoperative planning of brain surgeries. A virtual surgery system essentially consists of visualization and modeling of the medical data and deformation simulation of human organs (mainly soft tissues). We used the hybrid level set theory, threshold segmentation, and morphological methods to segment the human head MRI volumetric data into five parts, namely the outline of the head, gray matter, white matter, cerebrospinal fluid and ventricles. The iso2mesh toolkit based on Delaunay algorithm was then employed to generate the tetrahedral mesh and mark the five parts. We further studied the soft tissue deformation, and established a mechanic model based on Tensor-Mass system to simulate the point-touch and ball-touch models for soft tissue deformation. This approach can help to improve the positioning accuracy of a virtual brain surgery and represent the process of brain tissue deformation.

Identification of fresh and old vertebral compression fractures by MRI / 法医学杂志

OBJECTIVE@#To analyze and explore the value of MRI in distinguishing fresh from old vertebral compression fractures.@*METHODS@#The features of MRI in 43 cases with compression fractures of thoracic or lumbar vertebral bodies were analyzed. MRI sequences included T1WI, T2WI and STIR.@*RESULTS@#Fifty-five vertebral bodies in total were found compression fractures in 43 cases. Forty-six vertebral bodies, which showed low signals or low signals mixing a few high signals on T1WI, high signals on T2WI and significantly high signals on STIR, were identified as fresh compression fractures. Nine vertebral bodies were identified as old compression fractures, because they showed the same signals as normal vertebral bodies on T1WI, T2WI and STIR.@*CONCLUSION@#MRI could accurately distinguish fresh and old vertebral compression fractures, so it is valuable for the distinguishment in forensic identification.

Effects of decreased leptin expression on liver fibrosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the effects of decreased leptin expression on liver fibrosis.</p><p><b>METHODS</b>The small interfering RNA, targeting leptin gene, was designed according to the secondary structure of leptin gene. The recombinant plasmids were encapsulated with lipofectamine and then injected into carbon tetrachloride (CCl4) induced rat liver fibrosis models. Leptin and I, III collage were detected by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The mRNA and protein levels of leptin in the fibrotic liver transfected with leptin shRNA were significantly decreased compared with those in controls (P less than 0.01). The depositions of type I and type III collagens were also decreased (P less than 0.01).</p><p><b>CONCLUSION</b>Decreased leptin expression prevents liver fibrosis.</p>

Construction, expression and location of eukaryotic expression vector CMV-FLAG-RB in human prostate cancer PC-3 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct an eukaryotic recombinant expression vector for retinoblastoma 1 gene (RB-1) and investigate the role of RB-1 in prostate cancer.</p><p><b>METHODS</b>The coding sequence of RB-1 gene tagged with FLAG was amplified from the plasmid CMV-RB by PCR method. The fragment was cloned into CMV expression vector and identified by restriction enzyme digestion and sequence analysis. Western Blotting was used to detect RB-1 expression and immunofluorescence was used to observe RB-1 distribution in PC-3 cells transfected with the recombinant.</p><p><b>RESULTS</b>The expression vector CMV-FLAG-RB was successfully constructed as confirmed by PCR, endonuclease digestion and DNA sequence analysis. RB-1 protein was highly expressed and showed a nuclear distribution in PC-3 cells transfected with the recombinant.</p><p><b>CONCLUSIONS</b>The eukaryotic expression vector for RB-1 has been successfully constructed and can be efficiently expressed in PC-3 cells. The expression of RB-1 is located in the cell nuclei.</p>

Differential expressions of the receptor for advanced glycation end products in prostate cancer and normal prostate / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the differential expressions of the receptor for advanced glycation end products (RAGE) in the tissues of prostate cancer and normal prostate, and to find the role of RAGE in the pathogenesis of prostate cancer.</p><p><b>METHODS</b>We collected the tissue of prostate cancer and that of normal prostate from the same patient, and compared the differential expressions of RAGE at the tissue, protein and mRNA levels between prostate cancer and normal prostate tissues of 10 patients by immunohistochemistry, Western blot and real-time quantitative PCR.</p><p><b>RESULTS</b>Immunohistochemistry exhibited a significantly higher expression of RAGE in the prostate cancer tissue than in the normal prostate tissue; Western blot showed that the RAGE protein expression was 2.13 times higher in the former than in the latter (P < 0.05); and real-time quantitative PCR revealed the RAGE mRNA expression of the former to be 4.2 times that of the latter (P < 0.05).</p><p><b>CONCLUSION</b>RAGE may play an important role in the pathogenesis and progression of prostate cancer.</p>

Establishment of a high expressing system of human coagulant factor VIII in vitro / 中华血液学杂志

<p><b>OBJECTIVE</b>To construct a recombinant lentiviral vector (pXZ208-BDDhFVIII) mediating B-domain-deleted human coagulation factor VIII (BDDhFVIII) gene and investigate its expression in HLF, Chang-Liver and MSC cells.</p><p><b>METHODS</b>BDDhFVIII gene fragment was separated by endonuclease digestion and was cloned into the multiple cloning sites of pXZ208 to construct a recombinant lentiviral vector pXZ208-BDDhFVIII. Viral particles were prepared by means of three-plasmid cotransfection of 293T package cells by calcium phosphate precipitation. After infection, the coagulant activity of human FVIII in the culture medium of 293T, HLF, Chang-Liver and MSC cells was assayed by one-stage method. The gene transduction efficiency was assayed by flow cytometry (FCM). Furthermore, PCR was performed to test the integration of BDDhFVIII.</p><p><b>RESULTS</b>The infection rates of HLF, Chang-Liver and MSC were (74.52 +/- 7.57)%, (27.24 +/- 6.53)% and (42.34 +/- 5.84)% respectively. The activities of FVIII in supernatants of HLF, Chang-Liver and MSC were (54.1 +/- 5.6)%, (22.5 +/- 2.9)% and (12.5 +/- 2.7)% respectively. BDDhFVIII gene integration was detected in all the infected cells.</p><p><b>CONCLUSION</b>The recombinant lentiviral vector pXZ208-BDDhFVIII was successfully constructed and efficiently integrated into target cells to express human FVIII activity in vitro.</p>

Effect of bortezomib on acute graft-versus-host disease in mice model and its mechanism / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of bortezomib on prophylaxis of acute graft-versus-host disease (aGVHD) after mouse allogeneic-bone marrow transplantation (allo-BMT) and its mechanism.</p><p><b>METHODS</b>C57BL/6 (H-2(b)) mice were used as donors and BALB/c (H-2d+) mice as recipients. After allo-BMT, the BALB/c mice were divided into 3 groups, ie. group A:BMT control, group B: BMT + early infusion of bortezomib (1 mg kg(-1) d(-1), day 0-3), group C: BMT + late infusion of bortezomib (1 mg kg(-1) d(-1), day 5-7). Clinical manifestations of aGVHD, pathohistological changes, survival rate and levels of recipients H-2(b) cells detected by flow cytometry in the recipient mice were observed. Monodirectional mixed lymphocyte culture (MLC) system was established ex vivo and different concentrations of bortezomib (0, 2, 4, 8 nmol/L) were added to the system. The viability of the cells was detected by CCK-8 assay and cells apoptosis by flow cytometry. The concentrations of IL-2, IFN-gamma, TNF-alpha in the supernatant were detected by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The mice in group A developed typical aGVHD and all died of aGVHD within 3 weeks after transplantation, with a median survival time of (16.1 +/- 2.5) d. The symptoms of aGVHD was milder in group B than in group A, and the median survival time was significantly longer. The 60-day survival rate in group B was 70%, being significantly higher than that in other two groups(P<0.05). The mean value of donor-derived cell (H-2(b) cells) in group B was (98.1 +/- 1.1)% at 60 days. The symptoms of aGVHD was significantly severer in group C than in group A, and the median survival time was shorter. Bortezomib inhibited the cells viability in MLC system in a dose-dependent manner. After treated with 8 nmol/L bortezomib for 24 h, the inhibition ratio of cells viability was (41.4 +/- 6.0)%. The cell apoptosis rate increased gradually with bortezomib treatment for 12 h, 24 h and 36 h. After treated with 8 nmol/L bortezomib for 36 h, the apoptosis rate was (62.8 +/- 7.0)%. After treated for 24 h, the levels of IL-2, IFN-gamma and TNF-alpha in the supernatant were decreased.</p><p><b>CONCLUSIONS</b>Bortezomib administered immediately after allogeneic BMT can prevent aGVHD, improve the survival rate and have no influence of engraftment in the recipient mice. Delayed administration of bortezomib results in acceleration of aGVHD-induced mortality. Its mechanism maybe inhibition of the lymphocyte viability, increase of the cells apoptosis rate, and inhibition of secretion of IL-2, IFN-gamma, and TNF-alpha.</p>

Association of EGFR expression with angiogenesis and chemoresistance in ovarian carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To clarify the association of EGFR expression with angiogenesis and chemoresistance in ovarian cancer.</p><p><b>METHODS</b>Immunohistochemical PV-6000 staining was used to detect the expression of EGFR, LRP protein and MVD in 102 ovarian tumor specimens.</p><p><b>RESULTS</b>EGFR, LRP positive rates and MVD in borderline and malignant ovarian specimens were significantly higher than those in the normal and benign ones (P < 0.01). EGFR positive expression rate in stage III-IV carcinoma tissues, poor differentiation and with ascites was higher than that in stage I-II carcinomas of well differentiation and without ascites (P < 0.05). MVD was related to histological grade, residual tumor and ascites, LRP positive expression had no correlation with the clinicopathologic parameters (P > 0.05). The effective rate of chemotherapy in patients with EGFR and LRP-positive expression were 57.1% and 53.7%, respectively, significantly lower than that in cases with EGFR and LRP-negative expression (85.0% and 90.9%, P < 0.05). In the 64 cases with complete data, the three-year survival rate was 53.0%. The survival time was shorter in the cases with EGFR and LRP-positive expression, poor differentiation, ascites and chemoresistance (P < 0.01), and only LRP-positive expression and chemotherapeutic effect were independently related to survival time (P < 0.05). There was a correlation between EGFR and MVD (r = 0.548, P < 0.01), EGFR and LRP positive expression (P = 0.020).</p><p><b>CONCLUSION</b>The expression of EGFR in ovarian cancer is related to angiogenesis and chemoresistance. EGFR and LRP-positive expression are related to chemoresistance, and detection of the two proteins may be helpful in guiding chemotherapy choice for ovarian cancer. LRP-positive expression and chemotherapeutic effect may be independent prognostic factors.</p>