Effects of flurochloridone on apoptosis and Nrf2/HO-1 and NFκB signaling pathways in mouse testis and TM4 cells / 环境与职业医学

Background Flurochloridone (FLC) is toxic to male reproduction and can induce apoptosis of testicular tissue and supporting cells under oxidative stress. Of particular concern is whether nuclear factor-erythrocyte 2-related factor 2/heme oxygenase-1 (Nrf2/HO-1) signaling pathway and nuclear factor kappa B (NFκB) signaling pathway participate this process. Objective To observe apoptosis of testicular tissue and sertoli TM4 cells and alterations of Nrf2/HO-1 and NFκB signaling pathways in mice treated with FLC in vivo/in vitro. Methods (1) Animal experiment. Testis samples were harvested from male C57BL/6 mice after 28-day FLC (0, 3, 15, 75, and 375 mg·kg−1 per day) exposure via oral route. Malondialdehyde (MDA) and superoxide dismutase (SOD) in homogenate of testicular tissue were measured by colorimetry. Apoptosis of testicular tissue was evaluated by TUNEL staining. Expression and distribution of Nrf2 and NFκB were detected by immunohistochemistry. Protein expression levels of Nrf2, HO-1, NAD(P)H: quinone oxidoreductase 1 (NQO1), NFκB, inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ), and phosphorylated recombinant inhibitory subunit of nuclear factor kappa-B alpha (P-IκBα) in testicular tissue homogenate were determined by Western blotting. (2) Cell experiment. TM4 cell lines were treated with 40, 80, 120, 160, and 200 μmol·L−1 FLC for 6 h, and cell viability was detected by CCK-8. After 6 h exposure to 40, 80, and 160 μmol·L−1 FLC, the apoptosis rate was detected by flow cytometry, and the protein expression levels of Nrf2, HO-1, NQO1, NFκB, IKKβ, and IκBα were detected by Western blotting. Results (1) Animal experiment. Apoptosis occurred in the interstitial and basal parts of spermatogenic tubules in male C57BL/6 mice after 28 days of oral FLC exposure. Compared with the control group, the MDA level in testicular tissue of the 375 mg·kg−1 FLC-treated group was significantly increased (P<0.05), and the SOD activity was significantly decreased (P<0.05). After 375 mg·kg−1 FLC exposure, apoptosis occurred in the interstitial and basal parts of spermatogenic tubules. The results of immunohistochemistry showed the expression of Nrf2 and NFκB in the interstitium and basal part of spermatogenic tubules of the treated groups. Compared with the control group, the protein levels of Nrf2, NQO1, P-IκBα, NFκB, and IKKβ in the 15, 75, and 375 mg·kg-1 groups were significantly increased (P<0.001), and the HO-1 protein level was significantly increased in the 375 mg·kg−1 group (P<0.001). (2) Cell experiment. Compared with the control group, the TM4 cell viabilities in the 40, 80, 120, 160, and 200 μmol·L−1 FLC-treated groups significantly decreased (P<0.01). The apoptosis rates were significantly increased (P<0.05), and the apoptosis rates increased from 5.7% in the control group to 7.4%, 9.4%, and 11.7% in the 40, 80, and 160 μmol·L−1, respectively. The Nrf2 protein level in the 40 μmol·L−1 group was significantly increased (P<0.01), while the levels significantly decreased in the 80 and 160 μmol·L−1 groups (P<0.01). The HO-1 protein levels in the 40, 80, and 160 μmol·L−1 groups were significantly increased (P<0.01). The level of NQO1 protein in the 40 μmol·L−1 group was significantly increased (P<0.01). The NFκB protein levels were significantly increased in the 80 and 160 μmol·L−1 groups (P<0.001). The IκBα protein levels were significantly decreased in all treated groups (P<0.001). The IKKβ protein had no significant change. Conclusion FLC induces testicular tissue apoptosis, and the process affects Nrf2/HO-1 signaling pathway and NFκB signaling pathway. The in vitro study confirms that FLC could induce apoptosis of TM4 cells and activate Nrf2/HO-1 and NFκB signaling pathways.

RBMS3 Expression and Its Correlation with Clinicopathological Features and Prognosis of Epithelial Ovarian Cancer / 中山大学学报(医学科学版)

ObjectiveTo investigate the expression of RNA binding motif single stranded interacting protein 3 （RBMS3） in epithelial ovarian cancer （EOC） tissues and its relationship with the clinicopathological features and prognosis of EOC. MethodsThe study enrolled the paraffin-embedded tissues from 110 EOC cases and 73 benign epithelial ovarian tumor cases pathologically diagnosed in the first affiliated Hospital of Bengbu Medical College from January 2015 to December 2019. By using anti-RBMS3 polyclonal antibody， the immunohistochemical staining was employed to detect RBMS3 expression in the tissues and then its correlation with the clinicopathological parameters and prognosis of EOC was analyzed. ResultsRBMS3 was expressed in both EOC and benign epithelial ovarian tumor tissues. RBMS3 expression in EOC tissues， significantly related with International Federation of Gynecology and Obstetrics （FIGO） stage， histological grade， CEA levels and survival status， was significantly lower than that in benign epithelial ovarian tumor tissues （P<0.05）. Kaplan–Meier survival curve showed that low RBMS3 expression in EOC patients was correlated with decreased progression-free survival （PFS） and overall survival （OS） （P<0.05）. Univariate analysis showed that RBMS3 expression， FIGO stage， residual lesion size， intestinal metastasis and intraperitoneal implantation were associated with OS of EOC patients （P<0.05）； multivariate analysis showed that low RBMS3 expression and intestinal metastasis were independent risk factors for poor prognosis in EOC patients （P<0.05）. ConclusionsRBMS3 is expressed at low levels in EOC tissues， which is closely related to poor prognosis of EOC patients. RBMS3 may function as a tumor suppressor gene in EOC tissues and can be used as an EOC-independent prognostic marker for targeted therapy against EOC.

Inhibitory Effect of Cinobufotalin on Macrophage Inflammatory Factor Storm and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the inflammatory effects of Cinobufotalin on monocytes in resting state and macrophages in activated state and its molecular mechanism.@*METHODS@#THP-1 cells were stimulated with Phorbol 12-myristate 13-acetate to induce differentiation into macrophages. Lipopolysaccharides was added to activate macrophages in order to establish macrophage activation model. Cinobufotalin was added to the inflammatory cell model for 24 h as a treatment. CCK-8 was used to detect cell proliferation, Annexin V /PI double staining flow cytometry was used to detect cell apoptosis, flow cytometry was used to detect macrophage activation, and cytometric bead array was used to detect cytokines. Transcriptome sequencing was used to explore the gene expression profile regulated by Cinobufotalin. Changes in the significantly regulated molecules were verified by real-time quantitative polymerase chain reaction and Western blot.@*RESULTS@#1∶25 concentration of Cinobufotalin significantly inhibited the proliferation of resting monocytes(P<0.01), and induced apoptosis(P<0.01), especially the activated macrophages(P<0.001, P<0.001). Cinobufotalin significantly inhibited the activation of macrophages, and significantly down-regulated the inflammatory cytokines(IL-6, TNF-α, IL-1β, IL-8) released by activated macrophages(P＜0.001). Its mechanism was achieved by inhibiting TLR4/MYD88/P-IκBa signaling pathway.@*CONCLUSION@#Cinobufotalin can inhibit the inflammatory factors produced by the over-activation of macrophages through TLR4/MYD88/P-IκBa pathway, which is expected to be applied to the treatment and research of diseases related to the over-release of inflammatory factors.

Establishment of a Patient-Derived T-Cell Acute Lymphoblastic Leukemia Xenograft Model in Novel Immunodeficient NCG Mice / 中国实验血液学杂志

OBJECTIVE@#The leukemia cells from patients with T-cell acute lymphoblastic leukemia (T-ALL) were inoculated into NCG mice to establish a stable human T-ALL leukemia animal model.@*METHODS@#Leukemia cells from bone marrow of newly diagnosed T-ALL patients were isolated, and the leukemia cells were inoculated into NCG mice via tail vein. The proportion of hCD45 positive cells in peripheral blood of the mice was detected regularly by flow cytometry, and the infiltration of leukemia cells in bone marrow, liver, spleen and other organs of the mice was detected by pathology and immunohistochemistry. After the first generation mice model was successfully established, the spleen cells from the first generation mice were inoculated into the second generation mice, and after the second generation mice model was successfully established, the spleen cells from the second generation mice were further inoculated into the third generation mice, and the growth of leukemia cells in peripheral blood of the mice in each group was monitored by regular flow cytometry to evaluate the stability of this T-ALL leukemia animal model.@*RESULTS@#On the 10th day after inoculation, hCD45+ leukemia cells could be successfully detected in the peripheral blood of the first generation mice, and the proportion of these cells was gradually increased. On average, the mice appeared listless 6 or 7 weeks after inoculation, and a large number of T lymphocyte leukemia cells were found in the peripheral blood and bone marrow smear of the mice. The spleen of the mice was obviously enlarged, and immunohistochemical examination showed that hCD3+ leukemia cells infiltrated into bone marrow, liver and spleen extensively. The second and third generation mice could stably develop leukemia, and the average survival time was 4-5 weeks.@*CONCLUSION@#Inoculating leukemia cells from bone marrow of patients with T-ALL into NCG mice via tail vein can successfully construct a patient-derived tumor xenografts (PDTX) model.

From the body-mind holism to psychosomatic medicine: theory and practice of acupuncture and moxibustion / 中国针灸

The traditional life concept of "body-mind holism" in Chinese medicine has drawn a special attention with the development of modern psychosomatic medicine. The "body-mind holism" is valuable in the guidance for clinical acupuncture practice, but, it is only remained on the theoretical significance by the medical masters in the past dynasties. In the paper, based on the understanding of Huangdi Neijing (Yellow Emperor's Internal Classic), the similarities and differences are compared between the body-mind theory and psychosomatic medicine. In association with clinical observation, from three aspects, i.e. body-mind pathogenesis, acupoint detection and observing the mind, and body-mind treatment, the guidance of "body-mind holism" is explored for the clinical diagnosis and treatment of acupuncture and moxibustion; the referents of "body" and "mind" are clarified in views of disease, diagnosis and treatment; and the treatment strategies of acupuncture and moxibustion are discussed in the perspective of "body-mind holism".

Rapid identification of chemical constituents in Xuezhikang Capsule by UPLC-Q-TOF/MSE combined with the establishment of UNIFI MS database / 药学学报

An UPLC-Q-TOF/MSE technology coupled with UNIFI database was used to develop a rapid, high coverage, accurate and efficient chemical composition qualitative method for Xuezhikang Capsule. A UNIFI database was established utilizing compound name, formula, structure, following automatic matching with high-resolution mass numbers, isotope distributions, mass deviations, fragment ion matching, and chromatographic retention features in UNIFI database to achieve the qualitative results of natural products in Xuezhikang Capsules. Combined with manual confirmation, 82 chemical components were identified in Xuezhikang Capsules, and the MS2 fragmentation pathway of typical organic acids, flavonoids, monacrines, and monascus were analyzed to ensure accuracy of the LC-MS workflow. This study clarified the chemical substance basis of Xuezhikang Capsules by LC-MS technology, providing experimental data support for the identification of key quality attributes, quality control and consistency evaluation in the manufacturing process of Xuezhikang Capsules.

Physico-chemical and biological properties of different magnesium modified calcium phosphate bone cements / 中华创伤杂志

Objective:To investigate the physicochemical and biological properties of different magnesium modified calcium phosphate bone cements.Methods:The different magnesium modified calcium phosphate bone cements were divided into magnesium citrate, magnesium lactate, magnesium malate, magnesium phosphate and magnesium glycinate groups, each of which was added with different magnesium agents in the proportion of 0%, 1%, 3% and 5% of the total weight of calcium phosphate bone cements. The initial and final setting time, injectability, anti-collapse performance and compressive strength of different magnesium modified calcium phosphate bone cements were tested. Furthermore, the screened bone cement extracts were used to culture with third generation osteoblasts. Bioactivity assays were performed using the Cell Proliferation and Toxicity Assay Kit (CCK-8). Alkaline phosphatase (ALP) staining and Alizarin Red S (ARS) staining were performed on osteoblasts to observe the osteogenic activity of magnesium malate modified calcium phosphate bone cements.Results:The addition of different proportions of different magnesium agents led to the shortening of the initial and final setting time of modified calcium phosphate bone cements. Moreover, the final setting time of 5% magnesium malate modified calcium phosphate bone cements was the shortest (<40 minutes), which was significantly shorter compared with other magnesium agents in the same proportion (all P<0.05). With the addition of different magnesium agents in different proportions, the injectability of bone cements was gradually increased, and the injectability of 5% magnesium malate calcium phosphate bone cements reached the highest for (87.3±1.9)%, which was significantly increased compared with other magnesium agents in the same proportion (all P<0.05). The anti-collapse performance of bone cements was decreased with the addition of different magnesium agents in different proportions. Magnesium citrate, magnesium phosphate and magnesium glycinate modified calcium phosphate bone cements could not resist the flushing of deionized water. In particular, magnesium malate modified calcium phosphate bone cements had the best anti-collapse performance, with the maximum weight loss rate for only (9.8±2.3)% after 30 minutes of deionized water flushing, which was better than the rest of the groups (all P<0.05). The compressive strength of magnesium lactate and magnesium phosphate modified calcium phosphate bone cements showed a decrease compared with original calcium phosphate bone cements, while the compressive strength of magnesium citrate and magnesium malate modified calcium phosphate bone cements was significantly increased compared with original calcium phosphate bone cements, of which 3% magnesium malate modified calcium phosphate bone cements had the greatest compressive strength of (6.2±0.2)MPa, significantly higher than the rest of the groups (all P<0.05). The sieve test yielded magnesium malate modified calcium phosphate bone cement, which had a weight loss of (27.0±0.9)% at 35 days in vitro. The release of magnesium ions was increased with increasing magnesium malate dose in the in vitro environment of magnesium malate modified calcium phosphate bone cements in different ratios. A stable magnesium ion release was achieved within 35 days.Also, the pro-proliferative and osteogenic effects of modified calcium phosphate bone cements on osteoblasts were more obvious with increase of magnesium malate dose. For 5% magnesium malate modified calcium phosphate bone cements, the cell number, ALP staining area ratio and calcium nodule area ratio were significantly increased compared with the groups in the proportion of 0% and 1% magnesium malate (all P<0.05). Conclusions:Among magnesium citrate, magnesium lactate, magnesium malate, magnesium phosphate and magnesium glycinate modified calcium phosphate bone cements, magnesium malate modified calcium phosphate bone cements have relatively suitable setting time, excellent anti-collapse performance and mechanical strength. Meanwhile, 5% magnesium malate modified calcium phosphate bone cements have better biological activity among different ratios of magnesium malate modified calcium phosphate bone cements, suggesting a potential value for clinical application.

In vitro effect of a short hairpin RNA targeting epidermal growth factor receptor combined with sirolimus on proliferation and apoptosis of the human cutaneous squamous cell carcinoma cell line Colo-16 / 中华皮肤科杂志

Objective:To investigate the effect of a short hairpin RNA (shRNA) targeting epidermal growth factor receptor (EGFR) combined with sirolimus on proliferation and apoptosis of the human cutaneous squamous cell carcinoma cell line Colo-16, and to explore underlying mechanisms.Methods:Cultured Colo-16 cells were divided into 5 groups: normal cell group receiving conventional culture and treatment with phosphate-buffered saline (PBS) , negative control group transfected with a shRNA-NC-expressing plasmid and treated with PBS, sirolimus group receiving conventional culture and sirolimus treatment, EGFR shRNA group transfected with an EGFR shRNA-expressing plasmid and treated with PBS, and combined group transfected with an EGFR shRNA-expressing plasmid and treated with sirolimus. Methyl thiazol tetrazolium (MTT) assay was performed to evaluate cellular proliferative activity in the above groups from 24 to 96 hours, and flow cytometry to detect cell apoptosis after 48-hour treatment. Semiquantitative RT-PCR was conducted to determine the mRNA expression of Bcl-2 and Bax, and Western blot analysis to determine the expression of apoptosis-related proteins cleaved caspase-3, cleaved caspase-9, Bcl-2, Bax, cell proliferation-related proteins phosphorylated mammalian target of rapamycin (p-mTOR) , phosphorylated protein kinase B (p-AKT) , phosphorylated 70-kDa ribosomal protein S6 kinase (p-P70S6k) , and cyclin D1. Comparisons among groups were carried out by using one-way analysis of variance, and multiple comparisons between 2 groups by using Student-Newman-Keuls q test. Results:MTT assay showed that the proliferative activity of Colo-16 cells was significantly lower in the sirolimus group, EGFR shRNA group and combined group during 24 - 96 hours than in the normal cell group (all P < 0.05) , and higher in the combined group than in the sirolimus group and EGFR shRNA group at 24-96 hours (all P < 0.001) , and there was no significant difference in the cellular proliferative activity at any time points between the normal cell group and negative control group (all P > 0.05) . Flow cytometry showed that the apoptosis rate was significantly higher in the sirolimus group, EGFR shRNA group and combined group (9.52% ± 0.25%, 12.65% ± 0.23%, 19.81% ± 0.31%, respectively) than in the normal cell group (3.33% ± 0.18%, q = 60.07, 78.08, 122.81, respectively, all P < 0.001) and negative control group (3.42% ± 0.19%, q = 59.90, 77.91, 122.64, respectively, all P < 0.001) , and was highest in the combined group. As RT-PCR and Western blot analysis revealed, the sirolimus group, EGFR shRNA group and combined group showed significantly decreased mRNA expression of Bcl-2 and protein expression of cyclin D1, p-AKT, p-mTOR, p-P70S6K and Bcl-2, but significantly increased mRNA expression of Bax and protein expression of cleaved caspase-3, cleaved caspase-9 and Bax compared with the normal cell group (all P < 0.05) . Compared with the sirolimus group and EGFR shRNA group, the combined group showed significantly decreased mRNA expression of Bcl-2 and protein expression of cyclin D1, p-AKT, p-mTOR, p-P70S6K and Bcl-2 (all P < 0.05) , but significantly increased mRNA expression of Bax and protein expression of cleaved caspase-3, cleaved caspase-9 and Bax (all P < 0.01) . Conclusion:EGFR shRNA and sirolimus exerted a synergistic effect in inhibiting the proliferation and promoting the apoptosis of Colo-16 cells, which may be related to the inhibition of the phosphoinositide 3-kinase (PI3K) /AKT/mTOR pathway.

Comparison of postoperative visual quality between toric and non-toric multifocal intraocular lens implantation / 中华实验眼科杂志

Objective:To evaluate and compare the postoperative visual quality after phacoemulsification combined with toric (ART) and non-toric (ReSTOR) multifocal intraocular lens (IOL) implantation.Methods:A cohort study was conducted.Thirty-nine cataract patients (50 eyes) who underwent phacoemulsification combined with ART IOL implantation were enrolled as ART group, and 32 patients (41 eyes) who received ReSTOR IOL implantation were enrolled as ReSTOR group in Tianjin Medical University Eye Hospital from January 2017 to January 2018.Three months after surgery, the uncorrected distance visual acuity (UDVA), uncorrected intermediate visual acuity (UIVA) and uncorrected near visual acuity (UNVA), manifest refraction, defocus curve, contrast sensitivity (CS), modulation transfer function cutoff, Strehl2D ratio (SR), objective scattering index, OQAS values under 100%, 20%, 9% contrasts (OV 100%, OV 20%, OV 9%), total aberrations, total lower-order aberrations, total higher-order aberrations, spherical aberrations, coma and trefoil aberrations of the two groups were tested and compared.The study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Tianjin Medical University Eye Hospital (No.2019KY[L]-04). Written informed consent was obtained from each subject prior to entering the cohort.Results:UDVA at 3 months after surgery was (0.07±0.09)LogMAR and (0.09±0.12)LogMAR in ART group and ReSTOR group, and CDVA was (-0.01±0.07)LogMAR and (-0.01±0.07)LogMAR, and UIVA was (0.23±0.11)LogMAR and (0.22±0.13)LogMAR, and UNVA was (0.11±0.15)LogMAR and (0.06±0.11)LogMAR.UNVA was slightly better in ReSTOR group than ART group, and the difference was statistically significant ( t=2.085, P=0.040). The mean depth of focus was (4.12±0.79)D in ART group and (4.24±0.95)D in ReSTOR group.The postoperative residual astigmatism (0.32±0.31)D was significantly lower than preoperative corneal astigmatism (1.27±0.40)D in ART group ( t=13.209, P<0.001). CS values at 6, 12, and 18 c/d under photopic without glare, 12 and 18 c/d under photopic with glare, 3, 6, and 12 c/d under scotopic without glare in ART group were slightly lower than those in ReSTOR group, showing statistically significant differences (all at P<0.05) without clinical significance.SR and OV 20% values in ART group were 0.14±0.05 and 0.55±0.24, which were slightly lower than 0.17±0.06 and 0.66±0.29 in ReSTOR group, with statistically significant differences ( t=-2.012, P=0.048; t=-2.557, P=0.043). Total aberrations and coma aberration under 5 mm pupil diameter in ART group were 0.88(0.59, 1.13)μm and 0.21(0.13, 0.30)μm, which were higher than 0.58(0.47, 0.74)μm and 0.10(0.08, 0.21)μm in ReSTOR group, showing statistically significant differences ( Z=-2.073, P=0.038; Z=-2.101, P=0.036). Conclusions:Cataract phacoemulsification combined with ART IOL implantation can provide good vision and visual quality while correcting preoperative corneal astigmatism, and the resolution in dim light or low to medium spatial frequencies is slightly weaker than eyes implanted with ReSTOR IOL.

Consideration on the construction of acupuncture and moxibustion clinical diagnosis and treatment system with the core of "differentiation of disease location": taking "treatment" of Science of Acupuncture and Moxibustion as an example / 中国针灸

Based on the difference between disease and syndrome differentiation of acupuncture and moxibustion and each department of TCM, i.e. internal medicine, external medicine, gynecology and pediatrics, it is believed that "differentiation of disease location" is the key of disease and syndrome differentiation of acupuncture and moxibustion. According to the disease location, in the section "treatment" of Science of Acupuncture and Moxibustion, the disorders/illness are classified as zangfu disorder, disorders of limbs and meridians, dermatological disorder, ophthalmological and otorhinolaryngological disorder, gynecological disorder, as well as qi, blood and body fluid disorder. Besides, the rules of disease and syndrome differentiation, as well as treatment characteristics of each category are explained separately. It is considered that the clinical diagnosis and treatment system of acupuncture and moxibustion should be constructed by focusing on "differentiation of disease location".

Result analysis of serology test and NAT quantitative detection of HBV DNA reactive blood donors / 中国输血杂志

【Objective】 To investigate the related viral markers of HBV DNA reactive samples among voluntary blood donors in Shiyan area, so as to provide some reference for donor re-entry. 【Methods】 From August 2019 to June 2021, 78 samples with HBV DNA reactivity from voluntary blood donors in our blood station were collected, and they were further detected for HBV viral markers(using chemiluminescence assays), nucleic acid testing(NAT) of virus quantification and sequencing analysis. 【Results】 Forty-seven (60.3%)out of 78 HBV DNA reactive samples were from repeated blood donors, and 56.4%(44/78)of them were over 45 years old. Among the five viral markers of hepatitis B, 37.2%(29/78) were positive for anti-HBs alone, 19.2%(15/78) were positive for anti-HBe+ anti-HBc and anti-HBs+ anti-HBe+ anti-HBc, and 11.5% (9/78) were all items negative.A total of 62.8%(49/78) of samples were detected by NAT quantification and seven samples had been successfully sequenced for HBV. 【Conclusion】 NAT can effectively minimize the missed detection of HBsAg methodology and reduce the risk of HBV transmission through blood. The re-entry of HBV DNA reactive blood donors should be treated with care.

Clinical features of preterm infants with a birth weight less than 1 500 g undergoing different intensities of resuscitation: a multicenter retrospective analysis / 中国当代儿科杂志

OBJECTIVE@#To evaluate the clinical features of preterm infants with a birth weight less than 1 500 g undergoing different intensities of resuscitation.@*METHODS@#A retrospective analysis was performed for the preterm infants with a birth weight less than 1 500 g and a gestational age less than 32 weeks who were treated in the neonatal intensive care unit of 20 hospitals in Jiangsu, China from January 2018 to December 2019. According to the intensity of resuscitation in the delivery room, the infants were divided into three groups:non-tracheal intubation (@*RESULTS@#Compared with the non-tracheal intubation group, the tracheal intubation and ECPR groups had significantly lower rates of cesarean section and use of antenatal corticosteroid (@*CONCLUSIONS@#For preterm infants with a birth weight less than 1 500 g, the higher intensity of resuscitation in the delivery room is related to lower rate of antenatal corticosteroid therapy, lower gestational age, and lower birth weight. The infants undergoing tracheal intubation or ECRP in the delivery room have an increased incidence rate of adverse clinical outcomes. This suggests that it is important to improve the quality of perinatal management and delivery room resuscitation to improve the prognosis of the infants.

Role of the LRP1-pPyk2-MMP9 pathway in hyperoxia-induced lung injury in neonatal rats / 中国当代儿科杂志

OBJECTIVES@#To study the role of the low-density lipoprotein receptor-related protein 1 (LRP1)-proline-rich tyrosine kinase 2 phosphorylation (pPyk2)-matrix metalloproteinases 9 (MMP9) pathway in hyperoxia-induced lung injury in neonatal rats.@*METHODS@#A total of 16 neonatal rats were randomly placed in chambers containing room air (air group) or 95% medical oxygen (hyperoxia group) immediately after birth, with 8 rats in each group. All of the rats were sacrificed on day 8 of life. Hematoxylin and eosin staining was used to observe the pathological changes of lung tissue. ELISA was used to measure the levels of soluble LRP1 (sLRP1) and MMP9 in serum and bronchoalveolar lavage fluid (BALF). Western blot was used to measure the protein expression levels of LRP1, MMP9, Pyk2, and pPyk2 in lung tissue. RT-PCR was used to measure the mRNA expression levels of LRP1 and MMP9 in lung tissue.@*RESULTS@#The hyperoxia group had significantly higher levels of sLRP1 and MMP9 in serum and BALF than the air group (@*CONCLUSIONS@#The activation of the LRP1-pPyk2-MMP9 pathway is enhanced in hyperoxia-induced lung injury in neonatal rats, which may be involved in the pathogenesis of bronchopulmonary dysplasia.

Effect of calcium phosphate bone cement scaffolds loaded with emodin on osteogenic activity of osteoblasts / 中华创伤杂志

Objective:To explore the effect of calcium phosphate cement（CPC）scaffold loaded with emodin（EMO）on osteogenic activity of osteoblasts.Methods:The bone cement scaffold was prepared by mixing EMO powder and CPC powder（ratio 1∶9），adding citric acid and then was poured into polytetrafluoroethylene mold（EMO-CPC group）. A dose of 0.36 g CPC powder was mixed with citric acid and injected into the polytetrafluoroethylene mold（CPC group）. General morphology，setting time（initial setting time and final setting time），injection rate and compressive strength of stents were compared between the two groups. Primary osteoblasts were extracted and co-cultured with two sets of scaffolds. After co-culture for 3 days，their characterization was observed by scanning electron microscopy. Live/dead cell staining and 3-（4,5-dimethylthiazole-2）-2,5-diphenyltetrazolium bromide（MTT）colorimetric method were used to detect cell viability，toxicity and proliferation activity of scaffolds. Two sets of scaffolds were stained with immunofluorescence for osteopontin（OPN），and protein expression was observed under an inverted fluorescence microscope. After co-culture for 7 days，tetrazolium nitro blue/5-bromo-4-chloro- 3-indolyl-phosphate（NBT/BCIP）staining method was used for alkaline phosphatase（ALP）staining. After co-culture for 14 days，two sets of scaffolds were stained with Alizarin Red to detect their osteogenic activity.Results:Two sets of stents showed relatively smooth and flat topography under the scanning electron microscope. There were no significant differences in initial setting time，final setting time，injection rate and compressive strength of stents between two groups（ P > 0.05）. After co-culture for 3 days，the osteoblast clusters were adhered to the surface of the EMO-CPC scaffold，with good shape. Viable cell rate reached（98.2 ± 0.1）% in EMO-CPC group and（90.2% ± 0.1）% in CPC group（ P <0.05）. Cell proliferation activity in EMO-CPC group was stronger than that in CPC group（ P < 0.05）. OPN-specific staining showed that EMO-CPC group had stronger OPN protein fluorescence expression compared to CPC group. After co-culture for 7 days，expression of ALP in EMO-CPC group was higher than that in CPC group. After co-culture for 14 days，staining intensity of Alizarin Red in EMO-CPC group was more significant than that in CPC group. Conclusions:The EMO-CPC scaffold can provide a suitable environment for the growth of osteoblasts for it has better biocompatibility，cell proliferation and osteogenic activity than the CPC scaffold.

MiR-542-5p Inhibits Hyperglycemia and Hyperlipoidemia by Targeting FOXO1 in the Liver

Purpose@#This research was designed to investigate how miR-542-5p regulates the progression of hyperglycemia and hyperlipoidemia. @*Materials and Methods@#An in vivo model with diabetic db/db mice and an in vitro model with forskolin/dexamethasone (FSK/DEX)-induced primary hepatocytes and HepG2 cells were employed in the study. Bioinformatics analysis was conducted to identify the expression of candidate miRNAs in the liver tissues of diabetic and control mice. H&E staining revealed liver morphology in diabetic and control mice. Pyruvate tolerance tests, insulin tolerance tests, and intraperitoneal glucose tolerance test were utilized to assess insulin resistance. ELISA was conducted to evaluate blood glucose and insulin levels. Red oil O staining showed lipid deposition in liver tissues. Luciferase reporter assay was used to depict binding between miR-542-5p and forkhead box O1 (FOXO1). @*Results@#MiR-542-5p expression was under-expressed in the livers of db/db mice. Further in vitro experiments revealed that FSK/DEX, which mimics the effects of glucagon and glucocorticoids, induced cellular glucose production in HepG2 cells and in primaryhepatocytes cells. Notably, these changes were reversed by miR-542-5p. We found that transcription factor FOXO1 is a target of miR-542-5p. Further in vivo study indicated that miR-542-5p overexpression decreases FOXO1 expression, thereby reversing increases in blood glucose, blood lipids, and glucose-related enzymes in diabetic db/db mice. In contrast, anti-miR-542-5p exerted an adverse influence on blood glucose and blood lipid metabolism, and its stimulatory effects were significantly inhibited by sh-FOXO1 in normal control mice. @*Conclusion@#Collectively, our results indicated that miR-542-5p inhibits hyperglycemia and hyperlipoidemia by targeting FOXO1.

Effects of polypyramidine tract binding protein-associated splicing factor overexpression on apoptosis of human Müller cells under advanced glycation end products treatment / 中华眼底病杂志

Objective To observe the effect ofpolypyramidine tract binding protein-associated splicing factor (PSF) towards advanced glycation end products (AGEs) induced the apoptosis of Müller cells in vitro.Methods Experimental study.Müller cells were cultured and divided into groups according to the project design,plasmid enhanced green fluorescent protein-PSF were transfected into the cells to achieve the overexpression of PSF Müller cells in vitro,then cells were exposed to AGEs and the Morphological changes were observed by HE staining and Hoechst 33258 staining while the survival rate of cells were detected by MTT assay.The effects of PSF on AGEs-induced Müller apoptosis was measured by Cell Death Detection ELISA kit.Meanwhile,2',7'-diehlorofluorescin diaeetate staining was performed to monitor the protective effects of PSF on AGEs-induced Müller cells ROS.Results The morphology of cells in normal group was full and the cytoplasm staining was uniform.In N+AGEs group and Vec+AGEs group,cell volume decreased,cytoplasm was dense and concentrated,and eosinophilic staining was enhanced.The cell morphology of PSF+AGEs group was still full,with uniform cytoplasm staining and uniform nucleus staining.The viability of N+AGEs group,Vec+AGEs group and PSF+AGEs group were 0.42±0.11,0.35±0.12 and 0.68±0.12.The apoptosis values were 1.08 ± 0.16,0.96± 0.20 and 0.44± 0.08.The intracellular ROS levels were 28 833.67± 3 550.06,28 356.67±4 854.81,186 163.00±382.54.Compared with N+AGEs group and Vec+AGEs group,the cell viability of PSF+AGEs group was significantly improved (F=20.65,P=0.000),ce11 apoptosis value (F=43.43,P=0.000) and intracellular ROS level (F=1 8.86,P=0.000).Conclusion PSF overexpression play a protective role in AGEs-induced apoptosis by inhibiting the production of ROS in Müiller cells.

Improvement of education for clinical nutrition interns based on Joint Commission International standards / 中华医学教育探索杂志

Huashan Hospital became the first Academic Medical Center Hospital accredited by Joint Commission International (JCI) in 2013. The Department of Clinical Nutrition has constantly improved internship education through the introduction of JCI standards. Based on the flexible combination of teaching and practicing modules, clinical nutrition knowledge is closely integrated with professional dietitian skills, and in-depth practical training helps students to acquire much more experience of the occupation as a dietitian. Teachers think highly of students' initiative and knowledge conversion ability, and in the recent five years, undergraduate projects have been enhanced in both quantity and quality.

Improvement of education for clinical nutrition interns based on Joint Commission International standards / 中华医学教育探索杂志

Huashan Hospital became the first Academic Medical Center Hospital accredited by Joint Commission International (JCI) in 2013. The Department of Clinical Nutrition has constantly improved internship education through the introduction of JCI standards. Based on the flexible combination of teaching and practicing modules, clinical nutrition knowledge is closely integrated with professional dietitian skills, and in-depth practical training helps students to acquire much more experience of the occupation as a dietitian. Teachers think highly of students' initiative and knowledge conversion ability, and in the recent five years, undergraduate projects have been enhanced in both quantity and quality.

Experts consensus on the management of delirium in critically ill patients / 中华内科杂志

To establish the experts consensus on the management of delirium in critically ill patients.A special committee was set up by 15 experts from the Chinese Critical Hypothermia-Sedation Therapy Study Group.Each statement was assessed based on the GRADE (Grading of Recommendations Assessment,Development,and Evaluation) principle.Then the Delphi method was adopted by 36 experts to reassess all the statements.(1) Delirium is not only a mental change,but also a clinical syndrome with multiple pathophysiological changes.(2) Delirium is a form of disturbance of consciousness and a manifestation of abnormal brain function.(3) Pain is a common cause of delirium in critically ill patients.Analgesia can reduce the occurrence and development of delirium.(4) Anxiety or depression are important factors for delirium in critically ill patients.(5) The correlation between sedative and analgesic drugs and delirium is uncertain.(6) Pay attention to the relationship between delirium and withdrawal reactions.(7) Pay attention to the relationship between delirium and drug dependence/ withdrawal reactions.(8) Sleep disruption can induce delirium.(9) We should be vigilant against potential risk factors for persistent or recurrent delirium.(10) Critically illness related delirium can affect the diagnosis and treatment of primary diseases,and can also be alleviated with the improvement of primary diseases.(11) Acute change of consciousness and attention deficit are necessary for delirium diagnosis.(12) The combined assessment of confusion assessment method for the intensive care unit and intensive care delirium screening checklist can improve the sensitivity of delirium,especially subclinical delirium.(13) Early identification and intervention of subclinical delirium can reduce its risk of clinical delirium.(14) Daily assessment is helpful for early detection of delirium.(15) Hopoactive delirium and mixed delirium are common and should be emphasized.(16) Delirium may be accompanied by changes in electroencephalogram.Bedside electroencephalogram monitoring should be used in the ICU if conditions warrant.(17) Pay attention to differential diagnosis of delirium and dementia/depression.(18) Pay attention to the role of rapid delirium screening method in delirium management.(19) Assessment of the severity of delirium is an essential part of the diagnosis of delirium.(20) The key to the management of delirium is etiological treatment.(21) Improving environmental factors and making patient comfort can help reduce delirium.(22) Early exercise can reduce the incidence of delirium and shorten the duration of delirium.(23) Communication with patients should be emphasized and strengthened.Family members participation can help reduce the incidence of delirium and promote the recovery of delirium.(24) Pay attention to the role of sleep management in the prevention and treatment of delirium.(25) Dexmedetomidine can shorten the duration of hyperactive delirium or prevent delirium.(26) When using antipsychotics to treat delirium,we should be alert to its effect on the heart rhythm.(27) Delirium management should pay attention to brain functional exercise.(28) Compared with non-critically illness related delirium,the relief of critically illness related delirium will not accomplished at one stroke.(29) Multiple management strategies such as ABCDEF,eCASH and ESCAPE are helpful to prevent and treat delirium and improve the prognosis of critically ill patients.(30) Shortening the duration of delirium can reduce the occurrence of long-term cognitive impairment.(31) Multidisciplinary cooperation and continuous quality improvement can improve delirium management.Consensus can promote delirium management in critically ill patients,optimize analgesia and sedation therapy,and even affect prognosis.

Effects of Krüppel-like factor 6 overexpression towards apoptosis of human lens epithelial cells with ultra violetradiation B treatment / 中华实验眼科杂志

Objective To investigate the effect of the overexpression of Krüppel-like factor 6 (KLF6)towards the apoptosis of human lens epithelial cells (HLECs) induced by ultraviolet B (UVB) radiation.Methods The eukaryotic expression plasmid pEGFP-C2-KLF6 which was successfully constructed were transfected into HLECs,followed by the detection of KLF6 level by using Western blot,and then companied by UVB stimulation.Cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay.The morphology of the cells was observed by using hematoxylin-eosin staining method.The cell damage was examined by Live/Dead staining.The apoptotic markers bax and bcl-2 were detected by Western blot.Quantitative apoptotic levels were measured with the apoptosis detection kit;the expression level of reactive oxygen species (ROS) was analyzed by DCFH-DA probe.Results The cell viability of the 0.5 μg transfection group and the 1.0 μg transfection group was significantly lower than that of the blank vector control group (both at P＜0.05).In high KLF6 expression group,the cells were sparse,long and narrow in size and shape,and the cytoplasm was concentrated.The cells in the normal control group were green living cells with stable morphology and even quantity.The number of red dead cells was increased significantly in the KLF6 highexpression group.After UVB irradiation,the apoptosis value,relative bax expression,bax/bcl-2 ratio and ROS expression of HLECs cells in the KLF6 high-expression group were all higher than those in the blank vector control group,with statistically significant differences between them (all at P＜0.05).Conclusions Overexpression of KLF6 can exacerbate apoptosis of HLECs caused by UVB,by regulating the expression of apoptosis-related proteins and promoting the accumulation of ROS in the endoplasmic reticulum.Down-regulation of KLF6 expression by biological tools may play a protective role on LECs to a certain extent.