Dissection of Cellular Communication between Human Primary Osteoblasts and Bone Marrow Mesenchymal Stem Cells in Osteoarthritis at Single-Cell Resolution

Background and Objectives@#Osteoblasts are derived from bone marrow mesenchymal stem cells (BMMSCs) and playimportant role in bone remodeling. While our previous studies have investigated the cell subtypes and heterogeneity in osteoblasts and BMMSCs separately, cell-to-cell communications between osteoblasts and BMMSCs in vivo in humans have not been characterized. The aim of this study was to investigate the cellular communication between human primary osteoblasts and bone marrow mesenchymal stem cells. @*Methods@#and Results: To investigate the cell-to-cell communications between osteoblasts and BMMSCs and identifynew cell subtypes, we performed a systematic integration analysis with our single-cell RNA sequencing (scRNA-seq) transcriptomes data from BMMSCs and osteoblasts. We successfully identified a novel preosteoblasts subtype which highly expressed ATF3, CCL2, CXCL2 and IRF1. Biological functional annotations of the transcriptomes suggested that the novel preosteoblasts subtype may inhibit osteoblasts differentiation, maintain cells to a less differentiated status and recruit osteoclasts. Ligand-receptor interaction analysis showed strong interaction between mature osteoblasts and BMMSCs. Meanwhile, we found FZD1 was highly expressed in BMMSCs of osteogenic differentiation direction. WIF1 and SFRP4, which were highly expressed in mature osteoblasts were reported to inhibit osteogenic differentiation. We speculated that WIF1 and sFRP4 expressed in mature osteoblasts inhibited the binding of FZD1 to Wnt ligand in BMMSCs, thereby further inhibiting osteogenic differentiation of BMMSCs. @*Conclusions@#Our study provided a more systematic and comprehensive understanding of the heterogeneity of osteogenic cells. At the single cell level, this study provided insights into the cell-to-cell communications between BMMSCs and osteoblasts and mature osteoblasts may mediate negative feedback regulation of osteogenesis process.

Musk and carterii birdw enhance the effect of polygonum extract on chronic non-bacterial prostatitis: an animal experimental study / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the effects of musk and carterii birdw on the pathology and the expressions of inflammatory cytokines in chronic non-bacterial prostatitis (CNBP) rats treated with polygonum extract.</p><p><b>METHODS</b>Five male Wistar rats were used for the preparation of SC purified prostate protein solution, and another 48 randomly divided into four groups: polygonum extract, polygonum extract + musk and carterii birdw, CNBP model control and normal control. CNBP models were established by injecting SC purified prostate protein solution and Freund's complete adjuvant. At 60 days after modeling, the CNBP model control and normal control rats were treated with normal saline, and the other two groups with polygonum extract and polygonum extract + musk and carterii birdw, respectively (polygonum 1.575 g per kg per d, musk 0.021 g per kg per d, and carterii birdw 1.05 g per kg per d). After 14 days of continuous intragastric medication, all the rats were sacrificed for pathological examination, determination of the levels of TNF-alpha, IL-1beta, IL-6 and IL-8 in the prostate tissue homogenate by ELISA, and detection of the mRNA and protein expressions of inflammatory cytokines MCP-1 (CCL2) and CCR2 by RT-PCR and Western blot.</p><p><b>RESULTS</b>The polygonum extract + musk and carterii birdw group showed apparent improvement in the structure of the prostate tissue but no inflammatory infiltration, as was quite obvious in the polygonum extract group. Polygonum extract + musk and carterii birdw significantly decreased the inflammatory cytokines TNF-alpha ( [11.04 +/- 4.07] pg/ml), IL-1beta ([16.94 +/- 4.26] pg/ml), IL-6 ([110.08 +/- 28.42] pg/ml) and IL-8 ([26.28 +/- 7.36] pg/ml) in the prostate tissue, as compared with polygonum extract alone ([63.21 +/- 21.37] pg/ml, [41.32 +/- 14.62] pg/ml, [177.64 +/- 42.65] pg/ml and [96.37 +/- 37.61] pg/ml) (P < 0.05, P < 0.01). The former also exhibited significantly lower expressions of MCP-1 mRNA (0.32 +/- 0.17), MCP-1 protein (0.28 +/- 0.15), CCR2 mRNA (0.28 +/- 0.11) and CCR2 protein (0.11 +/- 0.04) than either the model control group (1.15 +/- 0.39, 0.93 +/- 0.34, 0.83 +/- 0.26 and 0.93 +/- 0.34) (P < 0.01), or the polygonum extract group (0.65 +/- 0.27, 0.56 +/- 0.22, 0.78 +/- 0.24 and 0.25 +/- 0.09) (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>Musk and carterii birdw can enhance the effect of polygonum extract on chronic prostatitis, reduce inflammatory response and improve tissue repair of the prostate in rats.</p>

Mutation screening and prenatal diagnosis of tuberous sclerosis complex / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To screen mutations of tuberous sclerosis complex (TSC) patients to confirm a clinical diagnosis of TSC, and to perform prenatal diagnosis for families with mutations.</p><p><b>METHODS</b>In this study, PCR-denaturing high-performance liquid chromatography(DHPLC), supplemented with sequencing when necessary, was used to screen TSC1 and TSC2 mutations in 21 patients from 19 pedigrees visited author's hospital in the last five years. For novel mutations, one hundred unrelated healthy individuals were screened to exclude the possibility of polymorphism.</p><p><b>RESULTS</b>Seventeen different mutations were found in 21 patients of 19 pedigrees with 13 being novel mutations, including c. 2672delA, c. 2672insA of TSC1 gene and c.4918insCGCC, c.1143delG, Intron27+1 G>A, c.1957-1958delAG, Intron5+1 G>A, c.910insCT, c.2753 C>G, c.4078dupAGCAAGTCCAGCTCCTC, Intron 11 -1 G>A, Intron 14+1 G>A, c.684 C>A of TSC2 gene, indicating a high frequency of de novo mutations in TSC. Three of these mutations were in the TSC1 gene (N762S, c.2672insA and c. 2672delA), while all remaining 14 were in the TSC2 gene. Prenatal diagnosis for TSC was performed for 7 fetuses from these pedigrees. The six fetuses that tested negative for TSC mutations were carried to term and, to date, none of these children has shown symptoms of TSC.</p><p><b>CONCLUSION</b>Author's data showed that a mutation detection rate of tuberous sclerosis was 89.5%(17/19) among patients in author's hospital. The ratio of TSC2 and TSC1 mutations was about 1:1 in the familial cases, but TSC2 mutation was more common than TSC1 mutation in sporadic cases. Author's data demonstrated that birth of TSC children for those with familial history of TSC could be prevented through prenatal diagnosis.</p>

Priming stimulation modifies synaptic plasticity in the perforant path of hippocampal slice in rat / 神经科学通报·英文版

Objective The potential of all central nervous system synapses to exhibit long term potentiation (LTP) or long term depression (LTD) is subject to modulation by prior synaptic activity, a higher-order form of plasticity that has been termed metaplasticity. This study is designed to examine the plasticity and metaplasticity in the lateral perforant path of rat. Methods Field potential was measured with different priming and conditioning stimulation protocols. Results Ten-hertz priming, which does not affect basal synaptic transmission, caused a dramatic reduction in subsequent LTP at lateral perforant path synapses in vitro, and the reduced LTP lasted for at least 2 h. The LTD was unaffected. The reduction of LTP in the lateral perforant path was also readily induced by applying priming antidromically at the mossy fibers. Conclusion Priming with 10 Hz, which is within a frequency range observed during physiological activity, can cause potent, long-lasting inhibition of LTP, but not LTD. This form of metaplasticity adds a layer of complexity to the activity-dependent modification of synapses within the dentate gyrus.

Effects of melatonin on the induction of long term potentiation in the hippocampal CA3 area of rats / 中国应用生理学杂志

<p><b>AIM</b>To study the effect of melatonin on the induction of LTP in CA3 area of hippocampus and to investigated its possible mechanisms.</p><p><b>METHODS</b>Melatonin and other drugs (Tacrine or DNQX) were microinjected into the CA3 area. By using extracellular electrophysiological recordings to observe the changes of the slope of fEPSP in the CA3 area.</p><p><b>RESULTS</b>(1) Evoked potential and the induction of LTP were depressed by different concentration of melatonin (0.2 microg/microl, 1 microg/microl and 5 microg/microl). As the melatonin concentration increased, the induction of LTP was blocked more obviously. (2) Melatonin could attenuate the excitation effect of Tacrine (inhibitor of AChE) on LTP. (3) Inhibition of the melatonin-induced on LTP attenuated by DNQX.</p><p><b>CONCLUSION</b>The application of melatonin in rats inhibits the induction of LTP in the hippocampal CA3 area. The action of melatonin on the induction of LTP may be through the modulation of not only non-NMDA receptors but also cholinergic system.</p>

Mutation screening and prenatal diagnosis of hidrotic ectodermal dysplasia in a Chinese family / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the mutations in Cx30 gene in a Chinese family with hidrotic ectodermal dysplasia (HED) and to make prenatal diagnosis on the embryo which has been pregnant for 5 months.</p><p><b>METHODS</b>A family including 2 affected and 4 unaffected individuals was collected, and their informed consents were obtained. The affected woman had a five-month pregnancy. An 884 bp fragment containing the whole GJB6 coding sequence was amplified by PCR and the products were bi-direction sequenced directly. The mutation was further confirmed with restriction endoenzyme digesting. On the base of successful gene diagnosis, the following detection procedure on the pregnant baby was performed. First the whole coding region of Cx30 was amplified using primers Cx30-F and Cx30-R and the PCR products were digested by Hae II. Then the PCR products were cloned into pUCm-T vector. Blue-white blot screening method and PCR-restriction endoenzyme digesting technique were used to identify the correct clones. The mutant allele clone was sequenced to confirmed the mutation.</p><p><b>RESULTS</b>A heterozygous missense mutation 263C --> T in the Cx30 gene was detected in the affected little girl and her affected mother, which led to an amino acid substitution (A88V) in the second transmembrane domain of GJB6. The mutation was confirmed by Hae II digestion. A88V mutant allele cannot be cut while the wild normal allele can be cut into two fragments, 520 and 278 bp. The result of analyse on the five-month pregnancy show the embryo carried the A88V mutation too. So the embryo will be a patient.</p><p><b>CONCLUSION</b>An A88V missense mutation in the Cx30 gene can also cause HED in Chinese Han population. Based on the gene diagnosis, prenatal diagnosis can be played using bi-direction sequencing and confirmed with restriction endoenzyme digesting.</p>

Analysis of 92 ectopic pregnancy patients after in vitro fertilization and embryo transfer / 中南大学学报(医学版)

OBJECTIVE@#To investigate the occurrence of ectopic pregnancy among women who received in vitro fertilization and assess the influential factors.@*METHODS@#The indications, methods of assisted conception and ectopic types were analyzed retrospectively after the patients received in vitro fertilization and embryo transfer (IVF-ET), intracytoplasmic sperm injection (ICSI), or freezing-thawing embryo transfer (FET).@*RESULTS@#A total of 6007 embryo transfers were performed, and 2322 (38.7%) clinical pregnancies were obtained. Ninety-four (4.05%) of them were ectopic pregnancies; and 92 were tubal pregnancies. The occurrence rate was 3.96%, which constituted 97.87% (92/94) of all ectopic pregnancies. There were 2 cases of other parts: one in abdominal cavity and the other in cornual pregnancy with the occurrence rate of 0.86%, constituting 2.32% (2/94). Twenty heterotopic pregnancies occurred (0.86%), constituting 21.28% (20/94). Among all ectopic pregnancies, the assisted conception of 86 cases was tubal pathology and/or pelvic adherence (91.49%), and 24 patients had a history of ectopic pregnancy (25.53%). The differences of clinical pregnancy rates between IVF-ET, ICSI and FET were not significant (P>0.05). The ectopic rate of IVF-ET group was significantly higher than that of ICSI or FET group (P<0.05), respectively. The ectopic rate in FET group was also higher than that in ICSI group (P<0.05).@*CONCLUSION@#The occurrence rate of ectopic pregnancy after IVF is higher than that of spontaneous pregnancy, and the main cause for ectopic pregnancy is the tubal pathological changes.

Prenatal diagnosis of common chromosomal aneuploidies on uncultured amniotic fluid cells by fluorescence in situ hybridization / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To evaluate the feasibility of using fluorescence in situ hybridization(FISH) for the detection of a few common chromosome aneuploidies on interphase nuclei of uncultured amniotic fluid cells.</p><p><b>METHODS</b>Amniotic fluid samples were taken from 55 women at 16-32 weeks of pregnancy; interphase FISH was performed for diagnosing Down syndrome and aneuploidies of other four chromosomes 13, 18, X and Y. Then the karyotypes from standard cytogenetic analysis after percutaneous umbilical blood sampling(PUBS) were compared to the FISH results.</p><p><b>RESULTS</b>Each of the 55 uncultured amniotic fluid samples tested with FISH was enumerated 200 nuclei. Fifty-three samples were normal. Two samples were found to have trisomy 21(one is a case of standard trisomy 21 with three signals in all 200 nuclei, the other is a mosaic trisomy 21).</p><p><b>CONCLUSION</b>Interphase FISH analysis of uncultured amniotic fluid cells is a rapid, accurate and very sensitive method. It could be used in the prenatal cytogenetic laboratory.</p>

Influence of PAF receptors on long term potentiation attenuated by aluminium in hippocampal CA3 area of rats / 中国应用生理学杂志

<p><b>AIM</b>To investigate the influence of platelet-activating factor (PAF) receptor on long-term potentiation (LTP) attenuated by aluminium.</p><p><b>METHODS</b>The method of extracellular recording was used to investigate the effect of PAF receptors on PP-CA3 LTP by microinjection of PAF receptor antagonist Ginkgolide B or agonist mc-PAF into CA3 area.</p><p><b>RESULTS</b>(1) Amplitude of population spikes (PS) of evoked potential was not affected but LTP induction was blocked by 0.2 micromol/L ginkgolide B in CA3 area. (2) LTP induction was not influenced by 0.25 mol/L aluminium chloride, however, it could be blocked when aluminium was applicated with ginkgolide B. (3) LTP induction was influenced slightly by 40 micromol/L mc-PAF but it has no difference in statistic. LTP induction could be blocked completely by 0.5 mol/L aluminium, but when aluminium was coapplicated with mc-PAF, this effect could be relieved.</p><p><b>CONCLUSION</b>These results indicate that PAF receptors are involved in induction of LTP in CA3 area by stimulating perforant path. The inhibitory effect of aluminium on LTP is partly related to PAF receptors.</p>