[Treatment status of tyrosine kinase inhibitor for newly-diagnosed chronic myeloid leukemia: a domestic multi-centre retrospective real-world study].

Objective: To retrospectively analyze the treatment status of tyrosine kinase inhibitors (TKI) in newly diagnosed patients with chronic myeloid leukemia (CML) in China. Methods: Data of chronic phase (CP) and accelerated phase (AP) CML patients diagnosed from January 2006 to December 2022 from 77 centers, ≥18 years old, and receiving initial imatinib, nilotinib, dasatinib or flumatinib-therapy within 6 months after diagnosis in China with complete data were retrospectively interrogated. The choice of initial TKI, current TKI medications, treatment switch and reasons, treatment responses and outcomes as well as the variables associated with them were analyzed. Results: 6 893 patients in CP (n=6 453, 93.6%) or AP (n=440, 6.4%) receiving initial imatinib (n=4 906, 71.2%), nilotinib (n=1 157, 16.8%), dasatinib (n=298, 4.3%) or flumatinib (n=532, 7.2%) -therapy. With the median follow-up of 43 (IQR 22-75) months, 1 581 (22.9%) patients switched TKI due to resistance (n=1 055, 15.3%), intolerance (n=248, 3.6%), pursuit of better efficacy (n=168, 2.4%), economic or other reasons (n=110, 1.6%). The frequency of switching TKI in AP patients was significantly-higher than that in CP patients (44.1% vs 21.5%, P<0.001), and more AP patients switched TKI due to resistance than CP patients (75.3% vs 66.1%, P=0.011). Multi-variable analyses showed that male, lower HGB concentration and ELTS intermediate/high-risk cohort were associated with lower cytogenetic and molecular responses rate and poor outcomes in CP patients; higher WBC count and initial the second-generation TKI treatment, the higher response rates; Ph(+) ACA at diagnosis, poor PFS. However, Sokal intermediate/high-risk cohort was only significantly-associated with lower CCyR and MMR rates and the poor PFS. Lower HGB concentration and larger spleen size were significantly-associated with the lower cytogenetic and molecular response rates in AP patients; initial the second-generation TKI treatment, the higher treatment response rates; lower PLT count, higher blasts and Ph(+) ACA, poorer TFS; Ph(+) ACA, poorer OS. Conclusion: At present, the vast majority of newly-diagnosed CML-CP or AP patients could benefit from TKI treatment in the long term with the good treatment responses and survival outcomes.

Surgical treatment of nasopharyngeal cancer - a consensus recommendation from two Chinese associations.

BACKGROUND: Surgical treatment is playing an increasingly important role in the management of nasopharyngeal carcinoma (NPC). This consensus focuses on the indications for optimal surgery, and surgical methods in the whole process of treatment for NPC to provide a useful reference to assist these difficult clinical decisions. METHODOLOGY: A thorough review of available literature on NPC and surgery was conducted by the Association for the prevention and treatment of nasopharyngeal carcinoma in China, international exchange and promotion Association for medicine and healthcare, and the Committee on nasopharyngeal cancer of Guangdong provincial anticancer association. A set of questions and a preliminary draft guideline was circulated to a panel of 1096 experienced specialists on this disease for voting on controversial areas and comments. A refined second proposal, based on a summary of the initial voting and different opinions expressed, was recirculated to the experts in two authoritative medical science and technology academic groups in the prevention and treatment of NPC in China for review and reconsideration. RESULTS: The initial round of questions showed variations in clinical practice even among similar specialists, reflecting the lack of high-quality supporting data and resulting difficulties in formulating clinical decisions. Through exchange of comments and iterative revisions, recommendations with high-to-moderate agreement were formulated on general treatment strategies and details of surgery, including indications and surgical approaches. CONCLUSION: By standardizing the surgical indications and practice, we hope not only to improve the surgical outcomes, but also to highlight the key directions of future clinical research in the surgical management of NPC.

[To compare the efficacy and incidence of severe hematological adverse events of flumatinib and imatinib in patients newly diagnosed with chronic phase chronic myeloid leukemia].

Objective: To analyze and compare therapy responses, outcomes, and incidence of severe hematologic adverse events of flumatinib and imatinib in patients newly diagnosed with chronic phase chronic myeloid leukemia (CML) . Methods: Data of patients with chronic phase CML diagnosed between January 2006 and November 2022 from 76 centers, aged ≥18 years, and received initial flumatinib or imatinib therapy within 6 months after diagnosis in China were retrospectively interrogated. Propensity score matching (PSM) analysis was performed to reduce the bias of the initial TKI selection, and the therapy responses and outcomes of patients receiving initial flumatinib or imatinib therapy were compared. Results: A total of 4 833 adult patients with CML receiving initial imatinib (n=4 380) or flumatinib (n=453) therapy were included in the study. In the imatinib cohort, the median follow-up time was 54 [interquartile range (IQR), 31-85] months, and the 7-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.2%, 88.4%, 78.3%, and 63.0%, respectively. The 7-year FFS, PFS, and OS rates were 71.8%, 93.0%, and 96.9%, respectively. With the median follow-up of 18 (IQR, 13-25) months in the flumatinib cohort, the 2-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.4%, 86.5%, 58.4%, and 46.6%, respectively. The 2-year FFS, PFS, and OS rates were 80.1%, 95.0%, and 99.5%, respectively. The PSM analysis indicated that patients receiving initial flumatinib therapy had significantly higher cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) and higher probabilities of FFS than those receiving the initial imatinib therapy (all P<0.001), whereas the PFS (P=0.230) and OS (P=0.268) were comparable between the two cohorts. The incidence of severe hematologic adverse events (grade≥Ⅲ) was comparable in the two cohorts. Conclusion: Patients receiving initial flumatinib therapy had higher cumulative incidences of therapy responses and higher probability of FFS than those receiving initial imatinib therapy, whereas the incidence of severe hematologic adverse events was comparable between the two cohorts.

[Clinical and genetic characteristics of young patients with myeloproliferative neoplasms].

Objectives: To investigate the clinical and genetic features of young Chinese patients with myeloproliferative neoplasms (MPN). Methods: In this cross-sectional study, anonymous questionnaires were distributed to patients with MPN patients nationwide. The respondents were divided into 3 groups based on their age at diagnosis: young (≤40 years) , middle-aged (41-60 years) , and elderly (>60 years) . We compared the clinical and genetic characteristics of three groups of MPN patients. Results: 1727 assessable questionnaires were collected. There were 453 (26.2%) young respondents with MPNs, including 274 with essential thrombocythemia (ET) , 80 with polycythemia vera (PV) , and 99 with myelofibrosis. Among the young group, 178 (39.3%) were male, and the median age was 31 (18-40) years. In comparison to middle-aged and elderly respondents, young respondents with MPN were more likely to present with a higher proportion of unmarried status (all P<0.001) , a higher education level (all P<0.001) , less comorbidity (ies) , fewer medications (all P<0.001) , and low-risk stratification (all P<0.001) . Younger respondents experienced headache (ET, P<0.001; PV, P=0.007; MF, P=0.001) at diagnosis, had splenomegaly at diagnosis (PV, P<0.001) , and survey (ET, P=0.052; PV, P=0.063) . Younger respondents had fewer thrombotic events at diagnosis (ET, P<0.001; PV, P=0.011) and during the survey (ET, P<0.001; PV, P=0.003) . JAK2 mutations were found in fewer young people (ET, P<0.001; PV, P<0.001; MF, P=0.013) ; however, CALR mutations were found in more young people (ET, P<0.001; MF, P=0.015) . Furthermore, mutations in non-driver genes (ET, P=0.042; PV, P=0.043; MF, P=0.004) and high-molecular risk mutations (ET, P=0.024; PV, P=0.023; MF, P=0.001) were found in fewer young respondents. Conclusion: Compared with middle-aged and elderly patients, young patients with MPN had unique clinical and genetic characteristics.

[A real world study on the relationship between drug resistance of targeted therapy and prognosis of HER-2-positive advanced breast cancer].

Objective: To explore the effect of primary and acquired resistance to anti-human epidermal growth factor receptor 2 (HER-2) on the overall survival of patients with HER-2 positive advanced breast cancer. Methods: The clinical characteristics of HER-2 positive patients with advanced breast cancer admitted to Cancer Hospital of Chinese Academy of Medical Sciences from January 1998 to December 2018 were collected, and their neoadjuvant/adjuvant and advanced three-line chemotherapy were summarized. Among them, targeted drugs for HER-2 included trastuzumab, pertuzumab, T-DM1, RC48-ADC, lapatinib, pyrotinib, allitinib, sipatinib, seratinib. Based on the duration of benefit from anti HER-2 treatment, the patients were divided into two groups: primary anti HER-2 resistance group and acquired anti HER-2 resistance group. In this study, the overall survival (OS) was used as the main end point. Kaplan-Meier analysis and Cox proportional risk regression model were used to analyze the effects of different drug resistance mechanisms on the overall survival. Results: The whole group of 284 patients were included. The median age of recurrence and metastasis was 48 years old, 155 (54.6%) were hormone receptor (HR) positive and 129 (45.4%) were HR negative, 128 cases (45.1%) were premenopausal and 156 cases (54.9%) were postmenopausal, 277 cases (97.5%) had a score of 0-1 in ECoG PS and 7 cases (2.5%) had a score of more than 2 in the first diagnosis of relapse and metastasis. There were 103 cases (36.3%) in the primary drug resistance group and 181 cases (63.7%) in the secondary drug resistance group. The median overall survival time of the two groups was 24.9 months and 40.4 months, respectively, with statistical significance (P<0.001). Conclusion: Primary resistance to HER-2 is one of the factors of poor prognosis in HER-2 positive breast cancer, and its mechanism needs to be further explored.

[Health-related quality of life and its associated variables in Chinese patients with Philadelphia-negative myeloproliferative neoplasms].

Objectives: To explore health-related quality of life (HRQoL) and identify its associated variables in Chinese patients with Philadelphia-negative myeloproliferative neoplasms (MPNs) . Methods: In this cross-sectional study, anonymous questionnaires were distributed to adult patients with MPNs to assess symptom burden measured by MPN-10 and HRQoL measured by Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (EORTC QLQ-C30) . Results: The data from 1405 respondents with MPNs, including 645 (45.9%) with essential thrombocythemia (ET) , 297 (21.1%) with polycythemia vera (PV) , and 463 (33.0%) with myelofibrosis (MF) , were analyzed. 646 (46.0%) respondents were male. The median age was 56 (range, 18-99) years. The mean MPN-10 scores were 13.0±12.7, 15.0±14.7, and 21.0±16.6 (P<0.001) , and the physical component summary (PCS) and mental component summary (MCS) scores were 48.0±8.5, 47.0±9.0, and 42.0±10.0 (P<0.001) and 51.0±11.0, 50.0±10.8, and 49.0±11.1 (P=0.002) for respondents with ET, PV, and MF, respectively. Respondents with MF reported the lowest score of physical functioning, role functioning, emotional functioning, cognitive functioning, social function, and global health status (all P<0.01) and the highest score of fatigue, pain, dyspnea, appetite loss, diarrhea, and financial problems (all P<0.05) in EORTC QLQ-C30. Multivariate analyses revealed that higher MPN-10 scores were significantly associated with lower PCS (-0.220 to -0.277, P<0.001) and MCS (-0.244 to -0.329, P<0.001) scores; increasing age (-1.923 to -4.869; all P<0.05) , lower PCS score. Additionally, comorbidity (ies) , symptom at diagnosis, splenomegaly, anemia, unknown driver gene, and higher annual out-of-pocket cost were significantly associated with lower PCS and/or MCS scores. However, age ≥ 60 years, urban household registration, concomitant medication, and receiving ruxolitinib therapy in respondents with MF were associated with higher MCS scores. Weak correlations were found between MPN-10 score (except the subscale of appetite loss and constipation) and EORTC QLQ-C30 score in majority of subscales in respondents with ET (|r| = 0.193-0.457, all P<0.001) , PV (|r| = 0.192-0.529, all P<0.01) , and MF (|r| = 0.180-0.488, all P<0.001) , respectively. Conclusions: HRQoL in patients with MPN was significantly reduced, especially in patients with MF. Sociodemographic and clinical variables were significantly associated with the HRQoL in patients with MPNs.

[Effect of integrated schistosomiasis control measures in Hunan Province from 2004 to 2019].

OBJECTIVE: To examine the effect of the integrated schistosomiasis control measures in Hunan Province from 2004 to 2019, so as to provide insights into the development of the schistosomiasis elimination strategy. METHODS: The integrated schistosomiasis control measures implemented by the health, agriculture, water resources, forestry, land and resources sectors were retrospectively collected in Hunan Province from 2004 to 2019, and the completion of each measure, cost of control measures, Schistosoma japonicum infections in humans and bovines, and snail status were analyzed each year. An index system for assessing the integrated schistosomiasis control effect was constructed using the Delphi method to calculate the integrated schistosomiasis control effect index. In addition, a cost-effect analysis was performed in terms of the decline in the prevalence of S. japonicum infections in humans and bovines, areas with snails in inner embankments, and areas with infected snails. RESULTS: A total of 7 110 926 500 Yuan were invested into the integrated schistosomiasis control program of Hunan Province from 2004 to 2019. During the 16-year period, a total of 277 437.12 hm2 snail habitats received molluscicidal treatments, 6 927 230 person-times given expanded chemotherapy, 2 116 247 bovine-times given expanded chemotherapy, 954 850 harmless toilets built, 290 359 bovines fenced, 136 666 bovines eliminated, 141 905 machines used to replace bovines, 39 048.63 hm2 water lands improved as dry lands, 724.12 km irrigation regions improved, 3 994 300 populations covered with safe water, 191 102.89 hm2 forests planted and 38 535.27 hm2 lands leveled. The prevalence of S. japonicum infections was 4.29% in humans and 4.48% in bovines in Hunan Province in 2004, with 2 449.37 hm2 snail habitats in inner embankments and 3 423.74 hm2 infected snail areas. In 2019, the prevalence of S. japonicum infections reduced to 0 in both humans and bovines, and areas of snail habitats reduced to 540.92 hm2 (77.92% reductions), while the areas with infected snails reduced to 0. The overall integrated schistosomiasis control effect index appeared a tendency towards a rise over years since 2004, and the integrated schistosomiasis control effect index was 97.35 in 2019; the annual mean costs for a 1% reduction in the prevalence of S. japonicum infections in 100 populations and 100 bovines were 70.11 Yuan and 4 204.78 Yuan, and the annual mean costs for a 1% reduction in the snail areas in inner embankments and infected snail areas were 2 010.20 Yuan and 1 298.09 Yuan, respectively. CONCLUSIONS: The integrated control measures achieve remarkable effectiveness for schistosomiasis control in Hunan Province, with a remarkable decline in the prevalence of S. japonicum infections in humans and bovines and great shrinking of snail areas in inner embankments and infected snail areas. Adequate fund investment is required to improve the integrated schistosomiasis control measures and consolidate the control achievements.

[Cervial vestibular-evoked myogenic potential induced by galvanic vestibular stimulation in normal people].

Objective:To establish a new method for detecting vestibular function by testing cervical vestibular-evoked myogenic potential induced by galvanic vestibular stimulation in normal population. Method:Twenty normal ears were tested for cervical vestibular evoked myogenic potential induced by galvanic vestibular stimulation. SPSS 18.0 software was used to analyze the obtained data. Result:In all healthy subjects mastoid-forehead galvanic vestibular stimulation produced a positive-negative biphasic EMG responses on SCM ipsilateral to the cathodal electrode. The latency of p13 was（11.52±3.05） ms. The latency of n23 was（15.31±3.38） ms. The amplitude of p13-n23 was（40.55±27.93） µV. The interval of p13-n23 was（3.53±1.38） ms. The interaural asymmetry ratio（AR, %） of p13, n23 latency, the amplitude and interval were respectively（6.96±6.79）%, （6.47±5.93）%, （28.08±26.42）% and （16.61±11.11）%. There was no significant difference in all parameters between the right and left ears of all subjects. Conclusion:The value of cervical vestibular-evoked myogenic potential induced by galvanic vestibular stimulation in normal subjects can be established to explore methods for diagnosis, treatment and researching mechanism of auditory neuropathy and vestibular neuropathy.

MicroRNA-577 inhibits cardiomyocyte apoptosis induced by myocardial infarction via targeting PARP1.

OBJECTIVE: The aim of this study was to investigate whether microRNA-577 could inhibit myocardial infarction (MI)-induced cardiomyocyte apoptosis by regulating poly ADP-ribose polymerase 1 (PARP1). MATERIALS AND METHODS: MI model was successfully established in mice by ligation of the left anterior descending coronary artery (LAD). The expression levels of microRNA-577 and PARP1 in myocardial tissues of mice were examined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). MI model in cells was established by hypoxia pre-treatment in primary cardiomyocytes and MCM cells. Subsequently, the expression levels of microRNA-577 and PARP1 in hypoxia preconditioning cardiomyocytes were determined as well. Meanwhile, Caspase3 activity in cardiomyocytes overexpressing microRNA-577 or PARP1 was detected using a relative commercial kit. Furthermore, the binding relationship between microRNA-577 and PARP1 was examined by Dual-Luciferase reporter gene assay. RESULTS: Infarct size/risk region and risk region/LV in MI group were (62.1±2.2)% and (57.6±1.9)%, respectively. Both of the above two indexes in the MI group were significantly higher than those of the control group. The serum level of LDH in MI mice increased by 2.8-fold when compared with controls. Meanwhile, the expressions of microRNA-577 and PARP1 in myocardial tissues of MI mice were markedly down-regulated in a time-dependent manner. Compared with normoxia preconditioning cardiomyocytes, microRNA-577 expression in hypoxia preconditioning MCM cells and primary cardiomyocytes was remarkably decreased. Dual-Luciferase reporter gene assay confirmed that microRNA-577 could bind to PARP1. After transfection of microRNA-577 mimics, the expression of PARP1 was significantly down-regulated. Moreover, microRNA-577 over-expression inhibited caspase3 expression in hypoxia preconditioning cells, which could be reversed by PARP1 up-regulation. Similarly, microRNA-577 over-expression markedly decreased infarct size, risk region and serum level of LDH in MI mice, which could be reversed by PAPR1 over-expression. CONCLUSIONS: MicroRNA-577 inhibits MI-induced cardiomyocyte apoptosis by degrading PARP1.

[Analysis on the distribution and drug resistance features of anaerobes isolated from clinical infectious specimens].

Objective: To clarify distribution and drug resistance characteristics of anaerobes isolated from clinical infectious samples, and to provide experimental data for guiding on treatment of infections caused by anaerobes. Methods: The anaerobes, isolated from 1 057 different clinical specimens from inpatients admitted to the Affiliated Hospital of Inner Mongolia Medical University from March 2016 to November 2017, were identified by VITEK-2 anaerobes and corynebacterium (ANC) card and bacteroides fragilis isolates were further verified by 16S-rRNA sequencing. Meanwhile, broth dilution method was employed to detect the drug sensititities of bacteroides fragilis and PCR method was used to detect the carbapenem resistance gene cfiA. Results: Totally 303 strains of anaerobic bacteria were isolated, among which 199 strains were gram-negative bacteria, accounted for 65.68%. Among the gram negative anaerobes, bacteroides species were the most common, accounted for 35.64%, followed by prevotella spp., which accounted for 19.14%. B. fragilis were the most common among Bacteroides spp., accounted for 21.54%, followed by Prevotella bivia, accounted for 5.94%. The coincidence rate of 16S-rRNA sequencing results of Bacteroides fragilis and that of ANC card identification results was 100% (65/65). The drug resistance of B. fragilis to penicillin, clidamycin, ampicillin/sulbactam, amoxicillin/clavulanic, were 100%, 90.77%, 56.92% and 66.67%, respectively. The resitance rate of B. fragilis to chloramphenicol was the lowest, which was 1.54%.Moreover, the resistance rate of B. fragilis to imipenem and metronidazole were both higher, which were 38.46%(25/65) and 23.08%(15/65), respectively, and the positive rate of beta lactamase was 100% (65/65). The carrying rate of carbapenems resistance gene cfiA for 65 strains of Bacteroides fragilis was 52.3% (34/65), and 72.00%(18/25)for imipenem resistant strains. Conclusions: The obligate anaerobic bacteria are widely distributed in clinical specimens, and the great majority are gram negative anaerobic bacteria, and the most frequently isolated one is Bacteroides fragilis. Bacteroides fragilis isolates are found to be resistant to several kinds of common antibiotics, especially for imipenem and metronidazole, which should be given more attention to.

Telomere-associated factor expression in replicative senescence of human embryonic lung fibroblasts.

The objective of this study was to find the key regulatory molecules in the cell senescence process through observing the expression of telomere-associated factor during the normal cell replicative senescence process. Based on the established cell replicative senescence model, reverse transcription-polymerase chain reaction and western blot analyses were used to detect telomere-associated factor expression at the mRNA and protein levels, including that of human telomere binding protein 1, tankyrase 1, telomerase RNA, telomere protection protein 1 (POT1), and p53 during the process of human embryonic lung fibroblast replicative senescence. The results showed that transcription of human telomere binding protein 1 did not change with cell senescence, whereas the protein expression of human telomere binding protein 1 increased gradually and then decreased rapidly; there was no change in the mRNA and protein expression of POT1; with the replicative senescence of human embryonic lung fibroblasts, expression of POT1 decreased gradually; TRF1 showed an increasing trend with cell senescence; and p53 protein expression did not change. Together, the results from this study suggest that human telomere binding protein 1, POT1, and TRF1 played important roles in cell senescence.

Clinical perspectives on mesenchymal stem cells promoting wound healing in diabetes mellitus patients by inducing autophagy.

BACKGROUND: The pathogenesis of wound healing in diabetes mellitus is complicated, and results in less effective healing. Mesenchymal stem cells (MSCs) have been thought to promote wound healing in diabetes. However, the underlying mechanisms are not well understood. Autophagy plays an important role in wound healing. It has been speculated that the mesenchymal stem cells derived from the umbilical cord can improve wound healing in diabetes mellitus by inducing autophagy. Hence, we reviewed the research progress in this field to identify new strategies of clinical treatment for wound healing in diabetes mellitus.

Molecular cloning and tissue distribution profiles of the chicken R-spondin1 gene.

Rspo1 belongs to the Rspo family, which is composed of 4 members (Rspo1-4) that share 40 to 60% sequence homology and similar domain organizations, and regulate the WNT signaling pathway via a common mechanism. Rspo1 plays a key role in vertebrate development and is an effective mitogenic factor of gastrointestinal epithelial cells. We report the cloning of chicken Rspo1 and its gene expression distribution among tissues. It contained an open reading frame of 783 bp encoding a protein of 260 amino acids, and its molecular weight was predicted to be 28.80 kDa. Reverse transcription-polymerase chain reaction-based gene expression analysis indicated that chicken Rspo1 was highly expressed in the stomach muscle tissue, but was expressed at low levels in the lung, brain, jejunum, cecum, ileum, spleen, pancreas, kidney, and glandular stomach. These results suggest that Rspo1 plays a major role in muscular immune protection.

Wnt3a promotes human umbilical cord mesenchymal stem cells to differentiate into epidermal-like cells.

OBJECTIVE: Mesenchymal stem cells are a population of pluripotent cells that can differentiate into epidermal-like cells under certain conditions. Wnt3a can promote the proliferation and differentiation of stem cells. However, the role of Wnt3a in the differentiation of human umbilical cord mesenchymal stem cells (hUCMSCs) into epidermal-like cells is unknown. MATERIALS AND METHODS: Third-generation hUCMSCs were cultured in normal medium, epidermal stem cell-conditioned medium, and conditioned medium with added Wnt3a. After culturing for 5 days, the expression of cytokeratin 19 (CK19), an antigen specific for epidermal-like cells, was assessed by immunofluorescence and flow cytometry. The expression of CK19 mRNA was confirmed by reverse transcription-polymerase chain reaction(RT-PCR), and ß-catenin expression was detected by western blot. RESULTS: hUCMSCs differentiated into epidermal-like cells when cultured in conditioned medium as shown by positive immunofluorescence staining for CK19. Flow cytometry showed that the number of cells positive for CK19 in the epidermal stem cell-conditioned medium group was significantly higher than that of control group, but lower than that of the Wnt3a-conditioned group (p < 0.05). RT-PCR showed that the expression level of CK19 mRNA in the conditioned medium group was significantly lower than that of the Wnt3a group (p < 0.01). Westernblots showed that the expression of ß-catenin in the conditioned medium group was significantly lower than that of the Wnt3a group (p < 0.01). CONCLUSIONS: These results suggest that Wnt3a can effectively promote the differentiation of hUCMSCs into epidermal-like cells.

SAHA attenuates sevoflurane-induced learning and memory impairments in fetal mice.

Previous studies have found that children with multiple exposures to anesthesia at an early age are at increased risk of learning and memory impairment. Sevoflurane is the most commonly used inhalational anesthetic for general anesthesia in children. Multiple exposures to sevoflurane have been shown to induce neuroinflammation, inhibit neurogenesis, and cause subsequent learning and memory impairments in fetal mice. Histone-tail acetylation has been implicated in memory formation. In this study, we employed suberanilohydroxamic acid (SAHA), an inhibitor of histone deacetylases, to treat sevoflurane-induced learning and memory impairments. Six-day-old C57BL/6 mice were exposed to sevoflurane for 2 h daily for 3 days. Morris water maze test performed to evaluate learning and memory impairments and the expression of genes related in to synaptic remodeling/plasticity, or regulated by neuronal activity or the cell cycle were detected by real-time PCR. We found that SAHA attenuated sevoflurane-induced learning and memory impairments in fetal mice. Our findings suggest that SAHA may have potential as a therapeutic agent for preventing or treating the neurotoxicity associated with anesthesia.

Therapeutic potential of umbilical cord mesenchymal stem cells with Wnt/ß-catenin signaling pathway pre-activated for the treatment of diabetic wounds.

The pathogenesis of diabetes mellitus wounds is complicate, and there lacks effective treatment strategies. Mesenchymal stem cells can promote wound healing. Compared with bone marrow mesenchymal stem cells, umbilical cord mesenchymal stem cells have obvious advantages in biological property. Wnts are potent regulatory molecules for stem cell turnover and skin regeneration, while Wnt signaling is not well activated in diabetic wounds. Umbilical cord mesenchymal stem cells with Wnt/ß-catenin signaling pathway pre-activated have some potential in the treatment of diabetic wounds. In this paper, we review the research status as well as problems in this field.

Epimorphin-induced differentiation of human umbilical cord mesenchymal stem cells into sweat gland cells.

OBJECTIVES: Mesenchymal stem cells (MSCs) have the potential for multi-directional differentiation and can be induced to differentiate into sweat gland cells under certain conditions. Epimorphin (EPM) plays an important role in the promotion of epithelial cell morphogenesis; however, its effect on sweat gland-cell differentiation of MSCs remains unknown. The purpose of this study was to investigate how EPM regulates sweat gland cell differentiation of human umbilical cord mesenchymal stem cells (hUCMSCs). MATERIALS AND METHODS: hUCMSCs were labeled with 5-bromo-2-deoxyuridine (BrdU) before differentiation induction; were cultured in common culture medium, conditioned medium, or EPM-conditioned medium; and then induced to differentiate into sweat gland cells. Five days after induction, the expression rates of the sweat gland-cell antigens cytokeratin 14 (CK14), cytokeratin 19 (CK19), and carcinoembryonic antigen (CEA) in hUCMSCs were detected by flow cytometry, and the messenger ribonucleic acid (mRNA) and protein levels of CK14, CK19, and CEA were determined by reverse transcription polymerase chain reaction (RT-PCR) and western blot, respectively. RESULTS: hUCMSCs can be induced to differentiate into sweat gland cells in conditioned medium, and expression of CEA was detected by immunofluorescence assay. Flow cytometry results showed that the expression rate of the sweat gland-cell antigens CK14, CK19, and CEA in the conditioned medium were significantly lower than that in the EPM conditioned medium (p < 0.05). RT-PCR and western blot results showed that the mRNA and protein levels of CK14, CK19, and CEA in the conditioned medium were all significantly lower than that in the EPM-conditioned medium (p < 0.01). CONCLUSIONS: These results suggest that EPM can effectively induce the differentiation of hUCMSCs into sweat gland cells.

Optimization of in vitro cell labeling methods for human umbilical cord-derived mesenchymal stem cells.

BACKGROUND AND OBJECTIVES: Human umbilical cord-derived mesenchymal stem cells (hUCMSCs) are a novel source of seed cells for cell therapy and tissue engineering. However, in vitro labeling methods for hUCMSCs need to be optimized for better detection of transplanted cells. AIM OF THE STUDY: To identify the most stable and efficient method for labeling hUCMSCs in vitro. MATERIALS AND METHODS: hUCMSCs were isolated using a modified enzymatic digestion procedure and cultured. hUCMSCs of passage three (P3) were then labeled with BrdU, PKH26, or lentivirus-GFP and passaged further. Cells from the first labeled passage (LP1), the fourth labeled passage (LP4) and later passages were observed using a fluorescence microscope. The differentiation potential of LP4 cells was assessed by induction with adipogenic and osteogenic medium. Flow cytometry was used to measure the percentage of labeled cells and the percentage of apoptotic or dead cells. The labeling efficiencies of the three hUCMSC-labeling methods were compared in vitro. RESULTS: BrdU, PKH26, and lentivirus-GFP all labeled LP1 cells with high intensity and clarity. However, the BrdU labeling of the LP4 cells was vague and not localized to the cell nuclei; LP9 cells were not detected under a fluorescence microscope. There was also a significant decrease in the fluorescence intensity of PKH26-labeled LP4 cells, and LP11 cells were not detected under a fluorescence microscope. However, the fluorescence of LP4 cells labeled with lentivirus-GFP remained strong, and cells labeled with lentivirus-GFP were detected up to LP14 under a fluorescence microscope. Statistical analyses indicated that percentages of LP1 cells labeled with PKH26 and lentivirus-GFP were significantly higher than that of cells labeled with BrdU (p < 0.05), and that the LP4 cells were more efficiently labeled with lentivirus-GFP than with PKH26 or BrdU (p < 0.05). BrdU-, PKH26-, and lentivirus-GFP labeled LP4 cells were all differentiated to adipocytes or osteoblasts with adipogenic and osteogenic medium. No statistical significance (p > 0.05) was observed between the death rates of labeled and unlabeled cells. CONCLUSIONS: Lentivirus-GFP is a valid method for long-term in vitro labeling, and it may be used as a long-term hUCMSC tracker following transplantation in vivo.

Microembolic signals predict cerebral ischaemic events in patients with moyamoya disease.

BACKGROUND AND PURPOSE: Recent studies found that microembolic signals (MESs) could be detected by transcranial Doppler in patients with moyamoya disease. However, the clinical significance of MESs in moyamoya disease remains unclear. Our aim was to investigate whether the MESs could predict cerebral ischaemic events in patients with moyamoya disease. METHODS: Fifty-four consecutive patients with moyamoya disease were recruited. MESs were monitored by transcranial Doppler for 30 min in the bilateral middle cerebral arteries of each patient on admission. Patients were followed up for 1 year. The primary end-point was cerebral ischaemic events including stroke and transient ischaemic attack (TIA). RESULTS: MESs were detected in 11 (20.4%) patients, with a frequency of 11 (10.2%) in 108 hemispheres. Logistic regression analysis revealed that previous ischaemic events within 3 months were associated with the presence of MESs (odds ratio 4.41, 95% CI 1.11-17.59). During a median follow-up of 384 days, 14 (13.0%) hemispheres had ischaemic events (seven strokes and seven TIAs). Cox regression showed that the hazard ratio for the risk of new ischaemic stroke and TIA in the hemispheres with MESs was 6.84 (95% CI 1.82-25.66) compared with those without, and 10.61 (95% CI 1.66-67.70) for ischaemic stroke alone, after controlling for age, sex, presence of ischaemic events at baseline, Suzuki stages and revascularization surgery. CONCLUSIONS: In patients with moyamoya disease, the presence of MESs is associated with recent ischaemic symptoms and independently predicts cerebral ischaemic events. MES detection may be of potential clinical value in the management of patients with moyamoya disease.

Preparation of microencapsulated VEGF gene-modified human umbilical cord mesenchymal stem cells and in vitro culture.

BACKGROUND AND OBJECTIVES: The application of microencapsulated stem cells has been shown to have many advantages in various fields of medical research. However, optimal modes for preparation of microencapsulate stem cells need to be improved, and expression and release of products of microencapsulated gene modified stem cells need to be studied in vitro. AIM OF THE STUDY: To explore the optimal parameters when preparing microencapsulated stem cells, and to investigate the effect of microencapsulation on growth, secretion, and metabolism of genetically modified human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs). MATERIALS AND METHODS: In this study, the parameters of preparation were regulated by observing the microcapsule shape and size. Live/dead cell viability kits and fluorescein isothiocyanate-labeled dextrans (FD) were used to detect the microencapsulated cell viability, and the permeability of microcapsules, respectively. Vascular endothelial growth factor (VEGF) production in the supernatant of microencapsulated and non-microencapsulated VEGF gene-modified hUCMSCs cultures was measured by ELISA. RESULTS: The optimal parameters of preparing microcapsules were regulated as followed: bolus velocity was 6 ml/h, and airflow velocity was 3 L/min. The morphology of microcapsules was a spherical structure with a diameter of 450 ± 30 µm. More than 90% of the cells were viable after 21 days of culture. Low and middle molecular weight FD was able to pass through the microcapsules; however, high molecular weight FD was not. Also, the VEGF concentration in microencapsulated and non-microencapsulated cell culture supernatants exhibited no significant difference at each time point. CONCLUSIONS: Microencapsulated stem cells can be ideally prepared via specifically regulated preparation. Lastly, microencapsulation does not alter growth, secretion, and metabolism of the genetically modified hUCMSCs.