Generation of an induced pluripotent stem cell line (KNIHi001-A) by reprogramming peripheral blood mononuclear cells isolated from a patient with Parkinson's disease.

Parkinson's disease is a degenerative brain disorder characterized by dopamine neuronal degeneration and dopamine transporter loss. In this study, we generated an induced pluripotent stem cell (iPSC) line, KNIHi001-A, from the peripheral blood mononuclear cells (PBMCs) of a 76-year-old man with Parkinson's disease. The non-integrating Sendai virus was used to reprogram iPSCs. iPSCs exhibit pluripotent markers, a normal karyotype, viral clearance, and the ability to differentiate into the three germ layers.

Trophoblast glycoprotein is a marker for efficient sorting of ventral mesencephalic dopaminergic precursors derived from human pluripotent stem cells.

Successful cell therapy for Parkinson's disease (PD) requires large numbers of homogeneous ventral mesencephalic dopaminergic (vmDA) precursors. Enrichment of vmDA precursors via cell sorting is required to ensure high safety and efficacy of the cell therapy. Here, using LMX1A-eGFP knock-in reporter human embryonic stem cells, we discovered a novel surface antigen, trophoblast glycoprotein (TPBG), which was preferentially expressed in vmDA precursors. TPBG-targeted cell sorting enriched FOXA2+LMX1A+ vmDA precursors and helped attain efficient behavioral recovery of rodent PD models with increased numbers of TH+, NURR1+, and PITX3+ vmDA neurons in the grafts. Additionally, fewer proliferating cells were detected in TPBG+ cell-derived grafts than in TPBG- cell-derived grafts. Our approach is an efficient way to obtain enriched bona fide vmDA precursors, which could open a new avenue for effective PD treatment.

Fine-tuning of dual-SMAD inhibition to differentiate human pluripotent stem cells into neural crest stem cells.

OBJECTIVES: The derivation of neural crest stem cells (NCSCs) from human pluripotent stem cells (hPSCs) has been commonly induced by WNT activation in combination with dual-SMAD inhibition. In this study, by fine-tuning BMP signalling in the conventional dual-SMAD inhibition, we sought to generate large numbers of NCSCs without WNT activation. MATERIALS AND METHODS: In the absence of WNT activation, we modulated the level of BMP signalling in the dual-SMAD inhibition system to identify conditions that efficiently drove the differentiation of hPSCs into NCSCs. We isolated two NCSC populations separately and characterized them in terms of global gene expression profiles and differentiation ability. RESULTS: Our modified dual-SMAD inhibition containing a lower dose of BMP inhibitor than that of the conventional dual-SMAD inhibition drove hPSCs into mainly NCSCs, which consisted of HNK+ p75high and HNK+ p75low cell populations. We showed that the p75high population formed spherical cell clumps, while the p75low cell population generated a 2D monolayer. We detected substantial differences in gene expression profiles between the two cell groups and showed that both p75high and p75low cells differentiated into mesenchymal stem cells (MSCs), while only p75high cells had the ability to become peripheral neurons. CONCLUSIONS: This study will provide a framework for the generation and isolation of NCSC populations for effective cell therapy for peripheral neuropathies and MSC-based cell therapy.

Off-the-Shelf, Immune-Compatible Human Embryonic Stem Cells Generated Via CRISPR-Mediated Genome Editing.

Human embryonic stem cells (hESCs) hold promise in regenerative medicine but allogeneic immune rejections caused by highly polymorphic human leukocyte antigens (HLAs) remain a barrier to their clinical applications. Here, we used a CRISPR/Cas9-mediated HLA-editing strategy to generate a variety of HLA homozygous-like hESC lines from pre-established hESC lines. We edited four pre-established HLA-heterozygous hESC lines and created a mini library of 14 HLA-edited hESC lines in which single HLA-A and HLA-B alleles and both HLA-DR alleles are disrupted. The HLA-edited hESC derivatives elicited both low T cell- and low NK cell-mediated immune responses. Our library would cover about 40% of the Asian-Pacific population. We estimate that HLA-editing of only 19 pre-established hESC lines would give rise to 46 different hESC lines to cover 90% of the Asian-Pacific population. This study offers an opportunity to generate an off-the-shelf HLA-compatible hESC bank, available for immune-compatible cell transplantation, without embryo destruction. Graphical Abstract.

Glial cells function as neural stem cells and progenitor cells / 生理学报

Glial cells, including astrocytes, oligodendrocyte progenitor cells (OPCs), NG2-glia, etc, are broadly distributed throughout the central nervous system (CNS). Also, it has been well known that glial cells play multi-roles in physiological and pathological processes in the CNS, such as maintaining homeostasis, providing neurotrophins for neurons and regulating neural signal transmission. Recently, increasing evidence showed that glial cells may also function as neural stem/progenitor cells and contribute to adult neurogenesis or neuroregeneration. In pathological conditions, for instance, astrocytes and OPCs could be activated to proliferate and differentiate. When cultured in vitro, they could form neurospheres which possess the ability to differentiate into astrocytes, oligodendrocytes and neurons. Additionally, forced expression of exogenous genes in astrocytes and NG2-glia can successfully reprogram them into neurons, which may also be suggestive of their stem/progenitor cell features. Here, we review current knowledge of the stem cell-like properties of glial cells, including what types of glial cells can function as stem/progenitor cells, how they can acquire the stem/progenitor potential and what progenies can be produced. These insights may foster a better understanding of glial cell biology and function in physiological or pathological processes in the CNS and lead to the idea of using the stem/progenitor-like glial cells as endogenous cell source for neural repair.

Astrocytes as therapeutic targets after spinal cord injury / 生理学报

Spinal cord injury (SCI) is a challenging medical problem in the field of neurology, showing high incidence rate, disability rate, treatment cost and low-aged trend. Despite the clinical application of drug intervention, surgical treatment and modern rehabilitation training, no ideal curative effect has been achieved. Therefore, future study is necessary to clarify detailed pathological mechanism of SCI and identify the potential target cells for therapeutic intervention. In the central nervous system (CNS), astrocytes are the most abundant and widely distributed glial cells which play multiple key roles in maintaining homeostasis of the CNS in physiological and pathological conditions. Increasing evidence indicates that astrocytes are ideal therapeutic target cells for SCI. Here, we review current knowledge of the roles of astrocytes in the pathological reaction after SCI, astroglial transplantation and astrocyte reprogramming.

Downregulation of chromobox protein homolog 7 expression in multiple human cancer tissues / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between chromobox protein homolog 7 (cbx7) expression and the occurrence and development of colorectal carcinoma (CRC), gastric carcinoma (GC) and hepatocarcinoma (HCC) tissues.</p><p><b>METHODS</b>The samples of neoplastic tissues and the corresponding cutting-edge normal tissues from 22 cases of CRC, 20 cases of GC, 30 cases of HCC were surgically collected. Level of cbx7 mRNA was detected with a fluorescent quantitative RT-PCR assay, and the correlationship among expression of cbx7 mRNA, the patients' clinicopathologic features and the surviving time after surgery was analyzed.</p><p><b>RESULTS</b>The relative copy number of cbx7 mRNA in carcinomas and the normal tissues was 0.010 ± 0.015 vs 0.053 ± 0.042 for CRCs, 0.197 ± 0.195 vs 1.891 ± 1.254 for GCs, and 0.008 ± 0.008 vs 0.030 ± 0.021 for HCCs, respectively. Compared with the corresponding normal tissues, cbx7 expression was significantly downregulated in CRCs, GCs, and HCCs (t = -7.351, -5.417 and -6.680, respectively, P < 0.01). The expression of cbx7 mRNA in CRCs had significant differences not only between two age groups (the relative copy number of cbx7 mRNA in age > 55 group was 0.007 ± 0.015, but 0.017 ± 0.012 in age ≤ 55 group, t = -2.586, P = 0.022); but also between vascular embolus-positive and negative groups (the level of cbx7 mRNA in positive and negative group was 0.022 ± 0.021 vs 0.006 ± 0.011, t = -3.175, P = 0.010). The area under the receiver operating characteristics (ROC) curve is 0.769 (P = 0.033). when the Cut-off value of the relative copy number of cbx7 mRNA was 0.002 in CRCs. The values less-than 0.002 were defined as low expression. The CRC patients with low expression of cbx7 had a shorter overall survival time; whose 5 years survival rate was only 30.8% (4/13); while the rate was 77.8% (7/9) in high expression of cbx7 group. The difference had statistical significance (χ(2) = 4.329, P = 0.037). The similar differences could not be found among GC and HCC patients.</p><p><b>CONCLUSION</b>Downregulation of cbx7 expression was very common among multiple carcinomas cases, and the downregulation influenced the prognosis of CRC patients.</p>

Case-control study on the treatment of Hawskins type II fractures of talar neck with minimally invasive or traditional approach / 中国骨伤

<p><b>OBJECTIVE</b>To compare the differences of radiographical and functional outcomes between traditional and minimally invasive approach in treating Hawskins type I fractures of the talar neck.</p><p><b>METHODS</b>From February 2000 to February 2008, 37 patients underwent reduction and internal fixation using two different approaches. In minimally invasive approach group, there were 16 males and 2 females, ranging in age from 20 to 47 years, averaged (35.2 +/- 3.1) years, and in traditional approach group, there were 15 males and 4 females, ranging in age from 22 to 44 years, averaged (35.3 +/- 2.3)years. All the patients had Hawskins type I fractures. The following data were compared between two groups: operation time, blood loss, length of incision and function of ankle joint. All the patients were graded by Hawkins points-scoring system after operation.</p><p><b>RESULTS</b>All patients were followed up. The duration ranged from 2 to 10 years, with an average of 5.6 years. The index such as operation time, blood loss and the length of incision of patients in minimally invasive approach group were significantly less than those of patients in traditional approach group. According to AOFAS points-scoring system, the score of minimally invasive group was higher than that of traditional approach group.</p><p><b>CONCLUSION</b>Compared with traditional approach, minimally invasive approach can protect blood supply better, get better clinical effects,which is worth of choosing to treat Hawskins type II fractures of talar neck.</p>

A new lignan from the seeds of Arctium lappa.

A new compound, neoarctin A (1), together with nine known compounds (2-10), were obtained from the ethanolic extract of the seeds of Arctium lappa. The structure of 1 was elucidated on the basis of spectral and chemical evidence.

Studies on chemical constituentsfrom leaves of Acer truncatum / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents from the leaves of Acer truncatum.</p><p><b>METHOD</b>Various chromatographic techniques were used to isolate and purify the constituents. The structures of these compounds were elucidated on the basis of spectral analysis.</p><p><b>RESULT</b>Seven compounds were isolated and identified as p-sitosterol (1), beta-amyrin (2), beta-amyrin acetate (3), 3, 5-dihydroxy-4-methoxybenzoic acid (4), astragalin (5), quercetin-3-O-beta-D-galactoside (6), and quercetin-3-O-alpha-L-rhamnoside (7).</p><p><b>CONCLUSION</b>All of compounds were isolated from this plant for the first time.</p>

Methylation specific binding activity of zinc finger protein Kaiso / 中华预防医学杂志

<p><b>OBJECTIVE</b>To compare binding activity of different zinc finger domain of human Kaiso with methylated CpG.</p><p><b>METHODS</b>pGEX constructs with different human Kaiso domain were generated and then corresponding fusion proteins were induced and purified. Electrophoretic mobility shift assays were applied to evaluate the binding activity of fusion proteins with methylated CpG.</p><p><b>RESULTS</b>The purified GST-KaisoZF fusion protein (without the POZ protein binding domain) could bind with methylated CpG probe specifically, but not for three or two zinc fingers without flanking domains.</p><p><b>CONCLUSION</b>Human zinc finger protein Kaiso could bind with methylated CpG specifically, only in the assistance of the neighboring flank sequence of the zinc finger domain.</p>

Effect of selenium-enriched garlic on chronic gastritis of the glandular stomach of Mongolian gerbils induced by H. pylori / 中华预防医学杂志

<p><b>OBJECTIVE</b>To detect the therapeutic effect of selenium-enriched garlic (SeG) on chronic gastritis.</p><p><b>METHODS</b>Chronic gastritis was induced of the glandular stomach of male Mongolian Gerbils via gastric instillation of H. pylori TN2 strain once every 4 days for 5 consecutive times followed by random classification into six groups. Fresh SeG suspension was administrated daily at dosages of 4.70, 1.5, 0.47, 0.15 g.kg(-1).d(-1) for four weeks. The gerbils in the positive control group were treated with omeprazole, clarithromycin, and amoxicillin for one week. The gerbils were killed for pathological examination four weeks after SeG-treatment.</p><p><b>RESULTS</b>Chronic gastritis (CAG), low-grade dysplasia or gastric intraepithelial neoplasia (DYS/GIN) were observed among 77% and 38.5% of the 13 H. pylori-treated animals in the negative control group, respectively; whereas 40% and 26.7% in the positive control group (n = 15), respectively. The incidences of CAG and DYS/GIN in the SeG groups (n = 21 - 27) were reduced dose-dependently, 16.7% - 38.7% and 11.1% - 14.3% for CAG and DYS/GIN, respectively.</p><p><b>CONCLUSION</b>SeG administration inhibits the development and progression of CAG induced by H. pylori remarkably.</p>

Study on the efficacy of hepatitis B virus vaccination and its influencing factors among children in rural area of Jiangsu province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of hepatitis B viruse (HBV) vaccination and its influencing factors among children in rural area of Jiangsu province.</p><p><b>METHODS</b>Twenty-five hundred and twenty-two children born after 1998 in rural area were selected as the study population using multistage cluster sampling method. HBsAg and anti-HBs were detected by enzyme linked immunoassay (ELISA) and radio-immunoassay (RIA), respectively. Anti-HBs negative children were boosted using different hepatitis B vaccines and the efficacy was compared. Factors causing HBV infection in HBsAg positive children were also investigated.</p><p><b>RESULTS</b>HBsAg positive rates in 1-7 year olds were 0.28%-1.28%, and the anti-HBs positive rates decreased from 76.7% to 45.5%. The HBsAg positive rate in children not timely vaccinated was significantly higher than those with HBV vaccine injection within 24 hours after birth (1.4% vs. 0.5%, P = 0.031). More than 90% of the anti-HBs negative children had protective level of anti-HBs after boosted with HBV vaccine.</p><p><b>CONCLUSION</b>HBsAg positive rate in children born after 1998 in rural area of Jiangsu province decreased significantly, with an average of 0.8%. The reason for HBsAg carriage in children might be attributed to mother-to-infant transmission or not timely HBV vaccination.</p>

Refinewent of extract of Sophora japonica L.by inorganic ceramic membrane / 中成药

AIM: To evaluate the effect of microfiltrating the extract of Sophora japonica L. by inorganic ceramic membrane. METHODS: The extract of Sophora japonica L. was processed by inorganic ceramic membrane, and the whole solid, effective ingredinent and flux of mebrane were examined. RESULTS: The extract of Sophora japonica L. became clear after microfiltration. The decreasing rate of the whole solid was 27.88%, the metastasis rate of the effective ingredient was 78.26, and the increasing rate of the effective ingredient was 8.48%. CONCLUSION: The microfiltration of inorganic ceramic membrane can improve the qualitry of extract of Sophora japonica L. refinery effect.