Effect of Human Bone Marrow Mesenchymal Stem Cells with Ectopic High OCT4 Expression on T Lymphocyte Function / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of human bone marrow mesenchymal stem cells (MSCs) with ectopic high OCT4 expression on T-cell proliferation, activation and secretion in vitro.@*METHODS@#Peripheral blood mononuclear cells were isolated from healthy children. Anti-CD3 and anti-CD28 monoclonal antibodies were used to activate T lymphocytes, which were stimulated by interleukin (IL)-2 for one week in vitro. Then MSCs with ectopic high OCT4 expression (MSC-OCT4) were co-cultured with activated T lymphocytes. After one week of co-culture, the supernatant was collected and the levels of Th1/Th2 cytokines [IL-2, IL-4, IL-6, IL-10, tumor necrosis factor (TNF)-α and interferon (IFN)-γ] were determined by flow cytometry. The lymphocytes after one week of co-culture were collected and counted by Countstar software. After the proportions of activated/inactivated T cell subsets were determined by flow cytometry, the absolute lymphocyte counts were calculated and expressed as mean ± standard deviation.@*RESULTS@#Compared with control T cell alone culture group, the proliferation of CD3+ T cells, CD3+CD4+ T cells, and CD3+CD8+ T cells were significantly inhibited in MSC group and MSC-OCT4 group. Compared with MSC, MSC-OCT4 could inhibit CD3+CD8+ T cell proliferation better (P =0.049), and mainly inhibited early T cell activation. Compared with control T cell alone culture group, the levels of IL-2 and INF-γ were significantly down-regulated both in MSC group and MSC-OCT4 group.After co-culture with T cells for one week, the level of IL-6 significantly increased in MSC group and MSC-OCT4 group compared with that before co-culture. Compared with control MSC group, MSC-OCT4 group had higher viable cell numbers after 1 week of co-culture (P =0.019), and could resist the inhibition of proliferation by higher concentration of mitomycin C.@*CONCLUSION@#Both MSC and MSC-OCT4 can inhibit the proliferation and activation of IL-2-stimulated T cells in vitro. After overexpression of OCT4, MSC has better proliferation ability in vitro and can inhibit the proliferation of CD3+CD8+ T cells more effectively, which may have a better and more lasting immunosuppressive ability to regulate the balance of Th1/Th2.

Effect of OCT4 Gene Expression Upregulation on the Expression of iPSC-related Transcription Factors in Human Bone Marrow Derived Mesenchymal Stem Cells / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of OCT4 over-expression on the expression of induced pluripotent stem cell (iPSC)-related transcription factors (cMYC，KLF4，LIN28，NANOG and SOX2) in human bone marrow derived mesenchymal stem cells (hBMMSCs), so as to provide fundamental basis for exploring the pathogenesis of hematological diseases (leukemia, aplastic anemia, etc.) from the perspective of hemopoietic microenvironment in the future.@*METHODS@#Recombinant plasmid pcDNA3.1-OCT4 was constructed and transferred into the optimal generation P3-4 hBMMSCs by liposome transfection. The cells with stable and high expression of OCT4（hBMMSCs-OCT4）were screened by G418 resistance screening (limited dilution) and subcloning, the expression of OCT4 mRNA and OCT4 protein was verified by RT-PCR and FCM, respectively. The expression of iPSC-related transcription factors (cMYC, KLF4, LIN28, NANOG and SOX2) were also determined by FCM and RT-PCR, so as to evaluate the effect of ectopic high expression of OCT4 on the expression of iPSC related transcription factors in hBMMSCs.@*RESULTS@#Recombinant plasmid pcDNA3.1-OCT4 was successfully constructed and cells with stable and high expression of OCT4 were successfully screened from hBMMSCs by limited dilution and subcloning. The result of flow cytometry showed that the mean expression level of OCT4 protein increased from (3.03±1.49)% to (95.46±1.40)% compared with the untransfected parental MSCs, which was also confirmed by RT-PCR analysis. At the same time, the expression levels of OCT4 protein and mRNA were compared between transient transfection (day 4) and stable expression cells (day 96), respectively, it was showed that the OCT4 protein level increased from (36.36±0.28)% at day 4 to (96.25±1.38)% at day 96, and the OCT4 mRNA level increased from 2.75-folds to 6.23-folds, respectively. Compared with the untransfected parental MSCs, the average expression levels of stemness transcription factors increased from (1.12±0.47)% (cMYC), (0.84±0.30)% (KLF4), (2.14±0.79)% (LIN28), (0.63±0.37)% (NANOG) and (14.34±2.44)% (SOX2) to (80.65±4.75)%, (73.03±4.70)%, (68.08±3.05)%, (39.39±1.85)%and (91.45±4.56)% in hBMMSCs-OCT4, respectively, which were consistent with results of RT-PCR analysis. Moreover, the expression levels of NANOG and SOX2 positively correlated with the mean expression of OCT4 (OCT4 vs NANOG: r=0.7802，OCT4 vs SOX2: r=0.4981；NANOG vs SOX2: r=0.7426).@*CONCLUSION@#Cells with stable and high expression of OCT4 have been successfully established from hBMMSCs. Ectopic high expression of transcription factor OCT4 in hBMMSCs can up-regulate the expression of other iPSC-related transcription factors such as cMYC, KLF4, LIN28, NANOG and SOX2.

HFE genetic variability and risk of alcoholic liver disease: A meta-analysis / 华中科技大学学报（医学）（英德文版）

Studies examining the association of hemochromatosis (HFE) gene polymorphisms and susceptibility to alcoholic liver disease (ALD) yielded inconsistent results. Thus, we performed a metaanalysis to investigate whether the variations in HFE gene increase the risk of ALD. The studies published up to Feb. 2014 were identified by searching PubMed/MEDLINE, ISI Web of Science, EMBASE and China National Knowledge Infrastructure databases, which was complemented by screening the references of the retrieved studies. For all genotypes and alleles, the odds ratios (ORs) with 95% confidence intervals (CIs) according to the heterogeneity were pooled using fixed-effect model. Sixteen studies with 1933 cases and 9874 controls were included for this meta-analysis. C282Y/C282Y, C282Y/wild type, H63D/wild type and C282Y/H63D were found not to be associated with susceptibility to ALD, but increased risk of H63D/H63D (OR: 1.52, 95% CI: 1.05-2.22, P=0.029) was observed for ALD when compared to total control. Comparison of ALD patients with alcoholics without liver damage revealed a significant association of D allele, as well as a marginal association of H63D/wild type with ALD, while H63D/H63D was not significantly associated with ALD although increased value of OR was obtained. The presence of Y allele and other genotypes yielded insignificant findings when ALD patients were compared with alcoholics without liver damage. No evident publication bias or significant heterogeneity among studies was detected in this meta-analysis. In conclusion, our metaanalysis showed a marginal higher prevalence of H63D variant in ALD but did not support an increased risk of C282Y mutation.

Visual detection of murray valley encephalitis virus by reverse transcription loop-mediated isothermal amplification / 生物医学与环境科学（英文）

A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of Murray valley encephalitis virus (MVEV) infection. The reaction was performed in one step in a single tube at 63 °C for 60 min with the addition of the hydroxynaphthol blue (HNB) dye prior to amplification. The detection limit of the RT-LAMP assay was 100 copies per reaction based on 10-fold dilutions of in vitro transcribed RNA derived from a synthetic MVEV DNA template. No cross-reaction was observed with other encephalitis-associated viruses. The assay was further evaluated using spiked cerebrospinal fluid sample with pseudotype virus containing the NS5 gene of MVEV.

Correction of secondary nasal deformity of cleft lip with autogenous costal cartilage framework / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the correction of secondary nasal deformity of cleft lip with autogenous costal cartilage framework.</p><p><b>METHODS</b>237 cases with secondary nasal deformity of unilateral cleft lip were treated. The rib cartilage was harvested through a mini-invasive incision, and was fabricated as a C-shaped framework, as well as some cartilage fragments. Through transcolumella incision, the C-shaped framework was implanted to support the depressed alar and the cartilage fragments were used to augment the nasal base.</p><p><b>RESULTS</b>Satisfactory cosmetic and functional results were achieved in all the patients with primary healing. 93 patients were followed up one year after operation with good cosmetic results.</p><p><b>CONCLUSIONS</b>Autogenous costal cartilage framework can be used for the correction of secondary nasal deformity of cleft lip with satisfactory results.</p>

Total nasal reconstruction with total rib cartilage framework / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate a new technique for nasal reconstruction with total rib cartilage framework.</p><p><b>METHODS</b>The expanded frontal flap was fabricated by skin expansion and flap delay to cover the reconstructed nose. The dorsal flap was reversed as the lining of reconstructed nose. The whole framework was made by rib cartilage. Secondary revision operation was also performed to make the reconstructed nose more natural.</p><p><b>RESULTS</b>Total nasal reconstruction was performed successfully in 37 cases. Each patient underwent 4-7 operation during a period of 6-8 months. 32 patients were followed up for 12-24 months. The reconstructed nose had a natural skin color and symmetric appearance with good ventilation and less scar. Both doctors and patients were satisfied with the results.</p><p><b>CONCLUSION</b>Satisfactory cosmetic result and ventilation function can be achieved by nasal reconstruction with total rib cartilage framework.</p>

Closure of cystic cavity-type bedsore by subcutaneous undermining dissection with continuous negative pressure drainage / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the clinical effect of subcutaneous undermining dissection with continuous negative pressure drainage for the closure of cystic cavity-type bedsore.</p><p><b>METHODS</b>12 patients with cystic cavity-type bedsore underwent surgical debridement and the wounds were closed after subcutaneous undermining dissection. The negative pressure drainage was put in the deep space. The healing process was observed.</p><p><b>RESULTS</b>Completed healing was achieved in all the 12 cases. The skin wounds healed after 17-20 days and the deep spaces closed after 36-43 days. 12 cases were followed up for 1 year with no occurrence.</p><p><b>CONCLUSIONS</b>It is an easy and effective method to treat cystic cavity -type bedsore by subcutaneous undermining dissection with continuous negative pressure drainage.</p>

Analysis of the 1000 Hz tympanometry in normal hearing neonates / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To analysis the clinical characteristics of 1000 Hz probe tone tympanometry obtained from normal neonates who passed the OAE screening. To calculate the normal range of the variances of the tympanometry, which may serve as the guide for newborn hearing screening and detect middle ear function in neonates.</p><p><b>METHODS</b>OAE screening were performed to screen the hearing with GSI-70 Automated OAE. The 1000 Hz probe tone tympanograms were obtained from 650 neonates who passed the TEOAE screening in both ears and were on the normal physiological conditions after birth with GSI Tympstar Version II Middle Ear Analyzer. The means, the standard deviation, the 95% confidence interval were analyzed.</p><p><b>RESULTS</b>The 1000 Hz tympanometric data showed the 1Y1B1G tympanogram in 732 ears (56.3%), the 1Y3B1G tympanogram in 145 ears (11.2%), the 0Y0B0G tympanogram in 269 ears (20.7%), other shapes in 154 ears (11.8%) according to the Vanhuyse model. The 1000 Hz tympanometric datas showed the single-peaked tympanogram in 967 ears, the 95% confidence interval of the tympanometric data were as follows: tympanometric peak pressure (Tpp) was from -55.0 to 180.0 daPa, peak compensated static acoustic admittance (Peak Ytm) was 0.03-1.18 mmHo, tympanometric width (TW) was 70.0-230.0 daPa.</p><p><b>CONCLUSIONS</b>The majority of 1000 Hz tympanograms from this study show the single-peaked in normal neonates. The 95% confidence interval of the tympanometric data may serve as a guide for hearing screening and detecting middle ear function in neonates.</p>

A quantitative study of anterior chamber angle with ultrasound biomicroscopy after cataract surgery with phacoemulsification and foldable intraocular lens implantation / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To study the changes in anterior chamber angle after phacoemulsification and foldable IOL implantation with ultrasound biomicroscopy.METHODS: Small-incision phacoemulsification and foldable IOL implantation were performed in 50 eyes of 46 senior patients, and the changes of anterior chamber angle were determined quantitatively by using ultrasound biomicroscopy before and one month after the surgery.RESULTS: In all patients, the angle was widened significantly one month after the surgery (P＜0.01). The measurements of TIA500 ( trabecular-iris angle at 500μm from the scleral spur) , AOD250 (angle-opening distance at 250μm from the scleral spur) and AOD500 (angle-opening distance at 500μm from the scleral spur) increased significantly after the operation ( P＜ 0.01). The mean post/pre-operative TIA500ratio, AOD250 ratio and AOD500 ratio were 1.65 (1.12-4.91),1.81 (1.06-2.67) and 1.65 (1.01-2.76), respectively. A significant negative correlation existed between preoperative and postoperative data.CONCLUSION: Small-incision cataract surgery deepens the anterior chamber and widens anterior chamber angle significantly in senior patients. The narrower the preoperative angle, the higher ratio of post/preoperative ratio found.

Evaluation of international classification criteria (2002) for primary Sjögren's syndrome in Chinese patients / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the sensitivity and specificity of international classification criteria (2002) for primary Sjögren's syndrome (pSS) and the role of lower lip biopsy in diagnosis of pSS in Chinese patients.</p><p><b>METHODS</b>Patients who were diagnosed by the experts/rheumatologists as pSS during 1990-2002 from the Department of Rheumatology, Peking Union Medical College Hospital were retrospectively collected as experimental group. Patients who were diagnosed as non-pSS connective tissue diseases or non-connective tissue diseases served as control group. Those with a history of head-neck radiation, hepatitis C virus infection, AIDS, lymphoma, sarcoidosis, graft versus host disease (GVHD), and anti-acetylcholine drug use were exempted. Both groups were required to complete questionnaires about symptoms such as dry eyes and dry mouth, and complete the objective tests of keratoconjunctivitis and xerostomia including Schirmer test, corneal staining, unstimulated salivary flow, sialography, lower lip biopsy, and antinuclear antibodies (including anti-SSA/SSB antibodies) test.</p><p><b>RESULTS</b>A total of 330 pSS patients were included in experimental group and 185 non-pSS patients in control group. The mean age of both groups matched (47.8 +/- 10.9 years vs. 46.2 +/- 13.6 years, P > 0.05). The sensitivities of the criteria in pSS patients with lower lip biopsy and in pSS patients without lower lip biopsy were 89.2% and 87.2%, respectively; the overall sensitivity was 88.5%. The specificity was 97.3%. A total of 11.3% pSS patients with negative anti-SSA/SSB anti- bodies were diagnosed as pSS by lower lip biopsy.</p><p><b>CONCLUSION</b>The international classification criteria (2002) for pSS is feasible in Chinese patients. It has high sensitivity and specificity, and may serve as diagnosis criteria in routine clinical practice.</p>