Taking Microbiota into Consideration in Mesenchymal Stem Cell Research.

Mesenchymal stem cells (MSCs) are a promising therapy in regenerative medicine, but the clinical efficacy has yet to be identified, because the functions of MSCs are modulated by many factors, including the age and health condition of donors, origin of the tissue, and several other unknown factors. Recently, it has been revealed that, besides host factors, the microbiota that inhabits the human body is a modulator of MSCs as well. Here, we highlight the role of microbiota in the alteration of MSCs functions, with a specific focus on the self-renewal ability, multiple differentiation potential, and the immunomodulation capacity of MSCs. We also review the clinical trials and model research on the synergic and antagonistic effects of microbiota in stem cell therapy. In addition, we discuss the underlying mechanisms of the interplay between microbiota and MSCs, which are elucidated using omics approaches followed by verification experiments. As oral and maxillofacial tissues are important sources of MSCs, as well as a major access to diverse microbes, further studies are needed to elucidate these interactions in the oral field to make greater advancements in regenerative medicine.

[Value of the simplified JSTH score criteria in the early diagnosis of sepsis-associated disseminated intravascular coagulation].

Objective: To evaluate the early diagnostic value of various indicators in the simplified JSTH score criteria for sepsis-associated disseminated intravascular coagulation (DIC). Methods: A retrospective study was conducted. Patients admitted to Intensive Care Unit (ICU) of the Second Affiliated Hospital of Kunming Medical University from January in 2017 to December in 2018 were enrolled. Totally of 365 patients were recruited, with 224 males and 132 females. The simplified JSTH score criteria was used to diagnose DIC. The patients were divided into sepsis with DIC group and sepsis without DIC group according to the diagnostic criteria of sepsis. Platelet (PLT), fibrin degradation products (FDPs), prothrombin time (PT), antithrombin (AT), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) scores, sequential organ failure assessment (SOFA) scores and the simplified JSTH scores were recorded on the first ICU day. Correlation analyses were conducted.Receiver operating characteristic (ROC) curves for diagnosis of DIC with each indicator were drawn to evaluate the diagnostic efficiency and predictive ability of 28-day mortality. Results: According to the simplified JSTH score, 143 cases of sepsis complicated with DIC were diagnosed. There were significant differences in PLT, FDP, AT, PT, APACHE Ⅱ score, SOFA score, 28-day mortality rate between the two groups (all P<0.01). It was shown by Pearson correlation analysis that the criteria has the best correlation with APACHEⅡ score and SOFA score (r=0.496 and 0.612, both P<0.01). The correlation between PLT and APACHE Ⅱ score or SOFA score was the best (r=-0.440 or-0.568, both P<0.01). It was shown by ROC curve that area under ROC curve (AUC) of PLT was 0.933, and the sensitivity and specificity was 93.0% and 85.0%, respectively. The 28-day mortality was predicted by using the indicators in the criteria. The AUG of AT was 0.813, and both the sensitivity (81.6%) and specificity (73.6%) were the highest. Conclusions: The simplified JSTH score criteria can be used for early diagnosis of sepsis-associated DIC and it is positively correlated with the severity of the disease. The correlation between PLT and the severity of disease is the best, and early diagnosis efficiency of PLT is the strongest. AT has a good predictive value for 28-day mortality.

Comparative transcriptome analysis of matched primary and distant metastatic ovarian carcinoma.

BACKGROUND: High grade serous ovarian carcinoma (HGSOC) is the most common subtype of epithelial ovarian cancers (EOC) with poor prognosis. In most cases EOC is widely disseminated at the time of diagnosis. Despite the optimal cytoreductive surgery and chemotherapy most patients develop chemoresistance, and the 5-year overall survival being only 25-35%. METHODS: Here we analyzed the gene expression profiles of 10 primary HGSOC tumors and 10 related omental metastases using RNA sequencing and identified 100 differentially expressed genes. RESULTS: The differentially expressed genes were associated with decreased embryogenesis and vasculogenesis and increased cellular proliferation and organismal death. Top upstream regulators responsible for this gene signature were NR5A1, GATA4, FOXL2, TP53 and BMP7. A subset of these genes were highly expressed in the ovarian cancer among the cancer transcriptomes of The Cancer Genome Atlas. Importantly, the metastatic gene signature was suggestive of poor survival in TCGA data based on gene enrichment analysis. CONCLUSION: By comparing the gene expression profiles of primary HGSOC tumors and their matched metastasis, we provide evidence that a signature of 100 genes is able to separate these two sample types and potentially predict patient survival. Our study identifies functional categories of genes and transcription factors that could play important roles in promoting metastases and serve as markers for cancer prognosis.

HMGA2 is associated with the aggressiveness of tongue squamous cell carcinoma.

OBJECTIVE: To analyze the effects of HMGA2 on proliferation, invasion, and metastasis in tongue squamous cell carcinoma (TSCC). METHODS: HMGA2 knockdown was performed in SCC15 cell lines, and functional assay was applied to observe the effects on cell migration and invasion. Real-time PCR, Western blotting, and immunohistochemistry (IHC) were also used to measure the expression of HMGA2 and EMT markers. RESULTS: HMGA2 expression was decreased after lentivirus infection. Functional assay showed that silence of HMGA2 can inhibit the proliferation of SCC15 cells and arrest the cells in G1/S phase. Moreover, knockdown of HMGA2 enhanced apoptosis of SCC15 cells. Wound-healing assay and transwell assay indicated that knockdown of HMGA2 significantly inhibited migration and invasion ability of SCC15 cells. Expression detection suggested that HMGA2 may be involved in the metastasis of SCC15 cells by activating Twist family expression and inducing epithelial-mesenchymal transition (EMT) process. IHC analysis showed that HMGA2 and vimentin were up-regulated in TSCC tissues, while E-cadherin was down-regulated. Clinicopathological analysis indicated that expression of HMGA2, E-cadherin, and Vimentin were associated with recurrence of patients with TSCC. CONCLUSION: Our findings demonstrated that HMGA2 may promote malignant transformation of TSCC through EMT process and may be an independent prognosis biomarker for TSCC.

Tauroursodeoxycholic Acid Attenuates Angiotensin II Induced Abdominal Aortic Aneurysm Formation in Apolipoprotein E-deficient Mice by Inhibiting Endoplasmic Reticulum Stress.

OBJECTIVE/BACKGROUND: Abdominal aortic aneurysm (AAA) is characterised by the infiltration of smooth muscle cell (SMC) apoptosis, inflammatory cells, neovascularisation, and degradation of the extracellular matrix. Previous work has shown that endoplasmic reticulum (ER) stress and SMC apoptosis were increased both in a mouse model and human thoracic aortic aneurysm. However, whether the ER stress is activated in AAA formation and whether suppressing ER stress attenuates AAA is unknown. METHODS: Human AAA and control aorta samples were collected. Expression of ER stress chaperones glucose-regulated protein (GRP)-78 and GRP-94 was detected by immunohistochemical staining. The effect of ER stress inhibitor tauroursodeoxycholic acid (TUDCA) on AAA formation in angiotensin (Ang) II induced apolipoprotein E-/- mice was explored. Elastin staining was used to observe the rupture of elastic fragmentation. Immunohistochemistry and Western blot analysis were performed, to detect the protein expression of ER stress chaperones and apoptosis molecules. RESULTS: There was significant upregulation of GRP-78 and GRP-94 in aneurysmal areas of human AAA and Ang II induced ApoE-/- mice (p < .05). TUDCA significantly attenuated the maximum diameters of abdominal aortas in Ang II induced ApoE-/- mice (p < .05). TUDCA significantly reduced expression of ER stress chaperones and the apoptotic cell numbers (p < .05). Furthermore, TUDCA significantly reduced expression of apoptosis molecules, such as caspase-3, caspase-12, C/EBP homologous protein, c-Jun N-terminal kinase activating transcription factor 4, X-box binding protein, and eukaryotic initiation factor 2α in Ang II induced ApoE-/- mice (p < .05). CONCLUSION: The results suggest that ER stress is involved in human and Ang II induced AAA formation in ApoE-/- mice. TUDCA attenuates Ang II induced AAA formation in ApoE-/- mice by inhibiting ER stress mediated apoptosis.

Spatial and temporal expression of histone demethylase, Kdm2a, during murine molar development.

The histone demethylase, lysine (K)-specific demethylase 2A (Kdm2a), is highly conserved and expressed ubiquitously. Kdm2a can regulate cell proliferation and osteo/dentinogenic, adipogenic and chondrogenic differentiation of mesenchymal stem cells (MSCs) derived from dental tissue. We used quantitative real-time RT-PCR analysis and immunohistochemistry to detect Kdm2a expression during development of the murine molar at embryonic days E12, E14, E16 and E17 and postnatal days P3 and P14. Immunohistochemistry results showed no positive staining of Kdm2a at E12. At E14, Kdm2a was expressed weakly in the inner enamel epithelium, stellate reticulum cells and dental sac. At E16, Kdm2a was expressed mainly in the inner and outer enamel epithelium, stratum intermedium and dental sac, but weaker staining was found in cervical loop and dental papilla cells adjacent to the basement membrane. At E17, the strongest Kdm2a staining was detected in the ameloblasts and stronger Kdm2a staining also was detected in the stratum intermedium, outer enamel epithelium and dental papilla cells compared to the expression at E16. Postnatally, we found that Kdm2a was localized in secretory and mature ameloblasts and odontoblasts, and dentin was unstained. Real-time RT-PCR showed that Kdm2a mRNA levels in murine germ cells increased from E12 to E14 and from E14 to E16; no significant change occurred at E16, E17 or P3, then the levels decreased at P14 compared to P3. Kdm2a expression may be closely related to cell proliferation, to ameloblast and odontoblast differentiation and to the secretion of extracellular enamel and dentin during murine tooth development.

Inducible nitric oxide inhibitor enhances the anti-tumor effect of cisplatin on CNE-2 cells by inducing cell apoptosis.

OBJECTIVE: Inducible nitric oxide (NO) synthase (iNOS) inhibitor S-methylisothiourea (SMT) has been reported to have anti-tumor effects on several types of cancers.  We aimed to investigate whether SMT can inhibit nasopharyngeal carcinoma cells CNE-2 proliferation through raise chemotherapy effect of diaminodichloroplatinum (DDP). MATERIALS AND METHODS: CNE-2 cells were treated with SMT, DDP and both of them respectively. MTT and colony-forming assay was performed to detect the proliferation effect of the treatment. Hoechst 33258 staining and apoptosis analysis were performed to investigate the apoptosis effect of chemotherapy. Additionally, the NO level was detected to estimate the activity of iNOS. RESULTS: CNE-2 cells expressed high level of iNOS. SMT can inhibit CNE-2 cells growth in a dose-dependent manner and have the effect on reducing dosage of DDP as well as enhancing the anti-tumor efficacy by promote cell apoptosis. CONCLUSIONS: Our findings suggested that SMT play a synergism role in the inhibition process of DDP on nasopharyngeal carcinoma, and SMT could be a promising therapeutic factor for cancer prevention.

Discrimination between Sjögren's and non-Sjögren's sicca syndrome by sialoscintigraphy and antibodies against alpha-fodrin and Ro/La autoantigens.

Both Sjögren's syndrome (SS) and non-Sjögren's syndrome (NSS) can present with the sicca symptoms of dry eyes and a dry mouth but they are distinct pathological entities that require diagnostic discrimination. This study included 82 sicca syndrome patients and examined the ability of sialoscintigraphy and antibodies against the autoantigens alpha-fodrin, Ro and La to discriminate between SS and NSS. A total of 30.8% of SS patients compared with 58.8% of NSS patients were alpha-fodrin positive. The prevalence of Ro positivity was 69.4% for SS patients compared with 0% for NSS patients. The prevalence of La positivity was 52.4% for SS compared with 0% for NSS patients. Sialoscintigraphy showed that more NSS patients had grade III salivary gland impairment compared with SS patients (64.7% versus 19.4%). These data suggest that using sialoscintigraphy in combination with measuring the levels of serum alpha-fodrin, Ro and La might be useful for SS and NSS discrimination.

Histone deacetylase-dependent transcriptional repression by pRB in yeast occurs independently of interaction through the LXCXE binding cleft.

We have developed a yeast model system to address transcriptional repression by the retinoblastoma protein (pRB). When fused to the DNA-binding domain of Gal4p (DB-pRB), pRB can repress transcription of reporter genes containing Gal4p binding sites; the histone deacetylase activity encoded by yeast RPD3 is required for DB-pRB repression. Mutation of the LXCXE binding cleft in pRB, a region reported to be required for histone deacetylase recruitment, does not interfere with pRB-mediated repression. From these findings based on yeast experiments, we surmise that the small pocket region of pRB must contain an additional domain that confers histone deacetylase-dependent transcriptional repression. This hypothesis was verified by experiments examining pRB-dependent histone deacetylase association in mammalian cells. In addition to RPD3, repression by pRB in yeast requires MSI1, an ortholog of RbAp48, but not SIN3 or SAP30. By comparing the genetic requirements of DB-pRB repression in yeast to those of other DB-repressor fusions, we can suggest a mechanism by which pRB recruits histone deacetylase activity.

Foix-Chavany-Marie syndrome.

Anarthria and bilateral central faciolinguopharyngeomasticatory paralysis with automatic voluntary dissociation are the clinical hallmarks of Foix-Chavany-Marie syndrome (FCMS), the cortico-subcortial type of suprabulbar palsy. We present a 64-year-old man who was admitted due to acute onset of speechlessness for days. Neurological examinations demonstrated typical features of FCMS. Cranial MRI showed a right opercular lesion and several lacunar infarcts in bilateral hemispheres. Hexamethylpropylenamine oxime (HMPAO) brain SPECT revealed hypoperfusion in bilateral opercular regions and notably on the right side. In review of previous reports, the most common etiology of FCMS is stroke in the region of either operculum. Our case may be a good example of FCMS after multiple cerebrovascular insults over bilateral opercular cortico-subcortical areas as demonstrated in MRI and SPECT studies.

Selective photoreceptor damage in albino rats using continuous blue light. A protocol useful for retinal degeneration and transplantation research.

PURPOSE: To develop a retinal degeneration model with selective photoreceptor loss and RPE sparing, to be used as recipient for evaluating retinal transplants. METHODS: Albino rats were exposed to blue light, continuously, for 1-7 days (24-168 h) in a specially designed cage. Eyes were histologically analyzed at periods between 2 h and 8 months after the light exposure. Electroretinograms (ERGs) were recorded from some rats at 12-216 days after exposure. Using behavioral methods, visual thresholds of some rats were determined before exposure and re-measured between 18 and 52 days following exposure. RESULTS: Apoptotic nuclei appeared exclusively in the photoreceptor layer after 1-5 days exposure to blue light. Light microscopy revealed that 2-4 days of light exposure reduced the outer nuclear layer (normally eight to ten rows) to 1 row of cells in the central retina and to two to three rows in the periphery, both in the superior and the inferior retina. Average ERG a- and b-wave amplitudes of light-damaged rats were both reduced by about 98%. Visual performance in the behavioral test was substantially impaired. CONCLUSIONS: Continuous exposure of albino rats to moderate blue light for 2-5 days selectively eliminates most of the photoreceptors while leaving the RPE initially intact.

Molecular cloning and analysis of Ca2+/calmodulin-dependent protein kinase II from the chicken brain.

The goals of this study were to identify specific mRNA for isoforms of calmodulin-dependent protein kinase II in chicken forebrain, prepare a cDNA expression library, and perform a sequence analysis of the kinase cDNA. Specific mRNAs for alpha- and beta-subunits of the kinase were identified in Northern blots. The mRNA for the alpha-subunit is larger in the chicken that in the rat, and for the beta-subunit is smaller in the chicken than the rat. Nucleotide sequencing of selected clones demonstrated the presence of an alpha-subunit with a 33 nucleotide insert known as the alpha-B-isoform. Clones of the beta-subunit showed it to contain a deletion of six nucleotides relative to previously described sequences. Variability in the mRNAs of calmodulin kinase II, as shown here, reflect the presence of species-dependent variability in gene structure as well as the presence of different functional isoforms.

Technetium-99m HIDA hepatic lobar distribution and retention ratios in detection of intrahepatic lithiasis.

Technetium-99m HIDA hepatic lobar distribution and retention ratios were developed to evaluate patients with intrahepatic lithiasis. The data of 57 cases were analyzed. Results reveal a highly significant difference in these ratios between the patients and normal individuals. They are simple, objective, and easily obtainable. Thus, the determination of these ratios may replace [99mTc]HIDA sequential scintiphotography, which is qualitative and time consuming, for screening intrahepatic lithiasis.

Sequential scintiphotography with technetium-99m pyridoxylideneglutamate in the detection of intrahepatic lithiasis: concise communication.

Sequential scintiphotography with Tc-99m pyridoxylideneglutamate was used to evaluate patients under strong suspicion of intrahepatic stones. The scintiphotos of 19 cases were analyzed. Results indicate that the sequential study reveals an excretion pattern pertinent to the diagnosis of intrahepatic stone. This simple and innocuous approach would be a useful screening technique for this disease.