Objective To explore the proliferation-promoting effect of bovine mammary gland epithelial cells (BMECs) co-cultured with umbilical cord mesenchymal stem cells (UC-MSCs) in serum-free culture mediuum.Methods Bovine UC-MSCs and BMECs were selected for co-culturing in direct or indirect contact.In the direct contact culture groups, UC-MSCs and BMECs were co-cultured at concentration ratios of 2∶1, 1∶1, 1∶2, 1∶3, 1∶4, 1∶5, and 1:10, respectively.In the indirect contact culture group, the supernatant of UC-MSCs was used as the conditioned medium to re-suspend BMECs.In the control groups, UC-MSCs and BMECs were cultured alone.The cell growth status in each group was observed at 0, 4, 8, 12, 24, 36, 48, 60, 72 h after culture, and cell proliferation was detected by cell counting kit-8 (CCK-8) assay.Results At 48 h, the optical density of the conditioned medium-BMECs group was significantly higher compared with the control groups (P<0.05).Meanwhile, the optical density in the direct contact group at a concentration ratio of 1∶2 reached the peak, which was extremely significantly higher compared with the control groups (P<0.01) and significantly higher compared with the other direct contact culture groups and the conditioned medium-BMECs group (P<0.05).Conclusions Co-culture of UC-MSCs and BMECs in serum-free culture medium is capable to promote the proliferation of BMECs, and the co-culture by cell-to-cell contact has a better effect.The optimal concentration ratio of UC-MSCs to BMECs is 1∶2, and the optimal culture time is 48 h.

Objective To analyze the clinical and nursing care of peripartum reversible posterior leukoencephalopathy syndrome (RPLS).Methods Thirty five cases with peripartum RPLS were retrospectively reviewed.Results All patients developed acutely,22.9％ in the antepartum period and 77.1％ in the postpartum period.The major clinical characteristics were headache (65.7％),seizures(62.9％),visual disturbances (54.3％),altered mental status (22.9％).The most common abnormality on neuro-imaging was edema predominantly involving the cortex and subcortical white matter in the posterior portions of the cerebral hemispheres,including the occipital lobes (77.1％),parietal lobes (65.7％),temporal lobes (28.6％) and frontal lobe (20.0％).MR diffusion-weighted imaging and ADC mapping revealed vasogenic edema.The treatments included aggressive blood pressure control and anticonvulsants with special nursing care.The clinical symptoms and the lesions on neuroimaging improved or resolved within 1 day to 2 months.Conclusions The prognosis for peripartum RPLS is ensured as long as the diagnosis is made properly and reasonable application of care program.

Objective To investigate the level of CD4+CD25+CD127low regulatory T cells (Tregs) in the peripheral blood of hepatocellular carcinoma (HCC) patients with HBV infection and to explore the relationship between these cells and HBV infection. Methods Peripheral blood mononuclear cells were isolated from 41 HCC patients infected with HBV, 34 HCC patients without HBV infection and 29 healthy persons. These mononuclear cells were labeled with the monoclonal anti-bodies of CD4, CD25 and CD127, and then Tregs level was determined by flow cytometry (FCM). Moreover, the load of HBV DNA in the blood serum of HCC patients with HBV infection was detected by fluorescent quantitative polymerase chain reac-tion (FQ-PCR). Results The percentage of CD4+CD25+CD127low Tregs in CD4+T cells in the HCC patients with HBV infec-tion was higher than that in the HCC patients without HBV infection [(8.24±2.20)%vs (7.06±2.46)%, P＜0.05] and that in the healthy persons [(8.24 ± 2.20)% vs (6.51 ± 1.99)%, P ＜ 0.01]. Furthermore, the percentage of CD4+CD25+CD127low Tregs in CD4+T cells in the HCC patients without HBV infection was higher than that in the healthy persons [(7.06±2.46)%vs (6.51± 1.99)%, P＜0.05]. In addition, the level of CD4+CD25+CD127low Tregs in the HCC patients with HBV infection was gradually increased following the increase of HBV DNA load and there was a positive correlation between the level of CD 4+CD25+CD127low Tregs and the load of HBV DNA (r=0.350,P ＜ 0.05). Conclusion The level of CD4+CD25+CD127low Tregs in HCC patients is increased remarkably and associated with the number of HBV DNA copy, which may be an important mecha-nism of immunologic escape of virus in HCC.

Objective To observe the effects of granulocyte colony stimulating factor (G-CSF) on electrophysiological properties of post-infarct ventricles.Methods Sixty-seven survival Wistar rats were divided into 4 groups:Sham group,Control group,MI early G-CSF group (E-G) and MI delay G-CSF group (D-G) after ligation of the left coronary artery as myocardial infarction model.Monophasic action potential(MAP) was recorded by absorption electrode in ex vivo perfused rat hearts.Effective refractive period(ERP),sinus cardiac length (SCL),action potential amplitude (APA),maximal depolariged (Vmax),ventricular fibrillation threshold(VFF) and ventricular fibrillation duration(VFD) were measured.Results The electrophysiological parameters (SCL,VFT,VFD,APA,ERP/MAP90,dispersion of ERP and MAP90) of the E-G group were improved significantly (all P ＜ 0.05) at day 7 post MI.Improvement in SCL,dispersion of ERP and MAP 90 were found in the D-G group as well at day 7 post MI (all P ＜ 0.05).Substained improvement in electrophysiological parameters were found in the E-G group at 3 months after MI (P ＜0.05).Besides SCL,APA,Vmax and dispersion of MAP90,all other parameters in the D-G group were similar to that of the control group with no statistical significance and even had a tendency of deterioration in ERP and MAP90 3 months after MI.Conclusion G-CSF intervention could improve electrophysiological properties of ischemic ventricles.Early G-CSF intervention showed better outcomes compared to delay G-CSF intervention on electrical remodeling ischemia myocardiumwhich may have effect on reducing the development of ventricular arrhythmia.

Objective To investigate the radiosensitizing effect of 17AAG-cypate micelles on human non-small cell lung cancer A549 cells and its possible mechanism.Methods (1) A single-hit multi-target model formula was used to analyze the radiosensitizing effects of 17AAG-M and 17AAG-cypate-M.(2) The effects of 17AAG-cypate-M on the viability of A549 cells under laser and X-ray irradiation were analyzed by MTT assay.(3) The effect of the drugs on the cell senescence was observed by β-galactosidase staining assay.(4) The effects of different treatment conditions on DNA damage repair were analyzed by γ-H2AX immunofluorescence staining assay.(5) The expression of p-Erk1/2 and p-Akt was measured by Western blot.The paired t test was used for analyzing the differences between groups.Results Compared with the X-ray irradiation group,the X-ray+17AAG-cypate-M group had a lower mean lethal dose and a sensitization enhancement ratio greater than 1,indicating that 17AAG-cypate-M had a radiosensitizing effect.Compared with the 17AAG-M group,the 17AAG-cypate-M group showed significantly lower cell viability (P<0.01),a significantly higher percentage of aging cells (P<0.01),and significantly further delayed DNA damage repair (P<0.01).And the 17AAG-cypate-M group had lower expression of p-Erk1/2 and p-Akt than the 17AAG-M group.Conclusions Compared with 17AAG-M,17AAG-cypate-M has a higher radiosensitizing effect on A549 cells.The mechanism might be inducing the cell senescence,delaying DNA damage repair,and inhibiting the expression of p-Erk1/2 and p-Akt.

AIM:To observe the effects of granulocyte colony-stimulating factor (G-CSF) on calcium, sodium and potassium ion channel currents of the ischemic atrial myocytes in guinea pig by whole-cell patch clamp technique.METHODS:The guinea pig atrial myocytes were obtained by enzymolysis.Under ischemia and hypoxia condition, whole-cell patch clamp was used to observe the effects of G-CSF at various concentrations on the changes of the I-V curve, activation curve and availability of L-type calcium channel current (ICa,L) and voltage-dependent sodium channel current (INa), as well as I-V curve of delayed rectifier potassium channel current (IK).RESULTS:Under ischemic condition, the I-V curves of ICa,L were changed by acute G-CSF intervention in a dose-dependent fashion.Except for G-CSF at dose of 300 μg/kg, the other concentrations of G-CSF did not change the activation curve and availability of ICa,L, indicating that the effects of G-CSF on ICa,L were in a voltage-independent fashion.The I-V curves of ICa,L under ischemic condition were gradually approaching the normal levels by the higher dose of G-CSF, while the effect of 300 μg/kg G-CSF on ICa,L was similar to 100 μg/kg G-CSF.Acute G-CSF intervention at different doses did not change I-V curve, activation curve, and availability or steady-state availability of INa.As a part of IK, the rapid activating component (IKr) was improved by 100 μg/kg and 300 μg/kg G-CSF intervention with the similar effects, while the slowly activating component (IKs) was not changed by G-CSF.CONCLUSION:G-CSF affects ion channel electrophysiological properties of ischemic atrial myocytes in a voltage-independent but concentration-dependent manner, thus reducing the incidence of atrial arrhythmia.

Objective: To investigate the effects of bone trauma therapy instrument combined with Guyuling capsule in the treatment of traumatic fractures and the effects on the levels of serum epidermal growth factor (EGF), transforming growth factor-1 (TGF-β1) and vascular endothelial growth factor (VEGF). Methods: The clinical data of 168 patients with traumatic fractures admitted to our hospital from January 2015 to December 2017 were retrospectively analyzed. According to different treatment methods, the patients were divided into the bone trauma treatment instrument group (54 cases), the bone Guyuling capsule group (52 cases) and the combined group (62 cases). All patients were treated with open internal fixation and routine anti-infection, detumescence, analgesic and other symptomatic treatment. The bone trauma therapeutic instrument group was given auxiliary treatment with bone trauma therapeutic instrument on the basis of conventional treatment, the bone healing capsule group was added with bone healing capsule treatment on the basis of conventional treatment, and the bone trauma therapeutic instrument and bone healing capsule combined treatment were given in the combined group at the same time. The clinical efficacy was compared and the serum levels of EGF, TGF-β1 and VEGF in the three groups were detected. Results: The detumescence time (8.37 ± 2.84 d vs. 10.76 ± 4.17 d, 11.64 ± 3.98 d, F=12.329) and the healing time (11.73 ± 3.44 week vs. 14.23 ± 4.62 week, 14.76 ± 5.17 week, F=7.835) of the combined group were significantly shorter than those of the bone trauma therapy group and the Guyuling capsule group (P<0.01). After treatment, the serum EGF (0.60 ± 0.27 μg/L vs. 0.75 ± 0.35 μg/L, 0.72 ± 0.37 μg/L, F=3.406), TGF-β1 (9.28 ± 4.19 μg/L vs. 12.36 ± 4.21 μg/L, 12.86 ± 4.69 µg/L, F=11.568), VEGF (92.58 ± 26.01 pg/ml vs. 132.69 ± 28.01 pg/ml, 127.63 ± 29.85 pg/ml, F=36.053) were significantly lower than those in the bone trauma therapy group and the Guyuling capsule group (P<0.01). Three months after operation, the cure rate was 35.2% (19/54) and the total effective rate was 81.5% (44/54) in all three groups. The cure rate and total effective rate in the combined group were better than those in the bone trauma therapy group and the Guyuling capsule group (χ²=6.373, 4.669, P=0.041, 0.031). Conclusions The treatment of traumatic fractures with bone trauma therapy instrument combined with Guyuling capsule can promote the healing of fractures, reduce the stress response caused by fractures, and improve the clinical efficacy.

Objective To investigate the CT features of renal oncocytoma(RO),and to analyze the causes of misdiagnosis.Methods CT and clinical data of 1 2 patients with RO confirmed by surgery and pathology were analyzed retrospectively,the CT features and the causes of misdiagnosis before operation were analyzed and summarized.Results According to CT features before operation,among the 1 2 cases of RO,9 were misdiagnosed as renal carcinoma,3 were considered as benign occupying lesions.There were 6 cases located in the left kidney and 6 in the right kidney.Seven cases showed round mass and 5 showed irregular mass.Plain CT showed homogeneous masses in 6 cases and heterogeneous masses in 6 cases.Enhanced CT showed masses with homogeneous enhancement in 2 cases and masses with progressive enhancement in 10 cases.The attenuation value of parenchymal enhancement ranged from 41 to 143 HU (mean 90.17 HU).Seven cases had central scar syndrome,of which 1 case had calcification in the scar.One case showed segmental enhancement inversion,8 showed conical interface and 5 had holding ball signs.Conclusion The CT features of RO is central scar syndrome, segmental enhancement inversion,conical interface and holding ball sign,etc,but the definitive diagnosis still rely on pathology.

OBJECTIVETo analyze horizontal transmission patterns of Streptococcus mutans among caries-active preschool children for early interventions of dental caries.

METHODSPlaque samples obtained from 20 caries-active preschool children between 4 and 5 years of age were cultured under anaerobic conditions for isolating S. mutans, which were identified by morphological and biochemical analyses and PCR using primers homologous to the surface protein glucosyltransferase B (gtfB). The genotypes of the isolated S. mutans strains were determined by arbitrarily primed PCR (AP-PCR).

RESULTSOf the 200 S. mutans isolates obtained, 19 were excluded by biochemical analysis, and the remaining 181 isolates were identified as S. mutans by PCR with primers of gtfB, showing 37 different genotypes as identified by AP-PCR. Six children were found to carry S. mutans of a single genotype, 11 carried 2 genotypes, 2 had 3 genotypes, and 1 had 4 genotypes; 2 children from different classes were found to carry S. mutans of the same single genotype.

CONCLUSIONWe identified 37 genotypes of S. mutans in these caries-active preschool children, among whom horizontal transmissions of the strains were not found.

Child, Preschool ; Dental Caries ; microbiology ; Dental Plaque ; Genotype ; Glucosyltransferases ; Humans ; Polymerase Chain Reaction ; Streptococcal Infections ; transmission ; Streptococcus mutans ; classification

OBJECTIVETo select and identify ssDNA aptamers specific to Streptococcus mutans strains with different cariogenicity isolated from clinical specimens.

METHODSSubtractive SELEX technology targeting the whole intact cells was used to screen for ssDNA aptamers specific to the clinical isolates Streptococcus mutans strains with different cariogenicity. Radioactive isotope, flow cytometry, gene cloning and sequencing, MEME online software and RNA structure analysis software were employed to analyze the first and secondary structures of the aptamers and identify the screened aptamers.

RESULTSDetection by radioactive isotope showed sufficient pool enrichment after 9 rounds of subtractive SELEX. Flow cytometry showed that the selected aptamers H1, H16, H4, L1, L10 and H19 were capable of binding specifically with highly cariogenic Streptococcus mutans strains but not with strains with a low cariogenicity. The aptamer H19 had the strongest binding capacity to highly cariogenic Streptococcus mutans strains, with a dissociation constant of 69.45∓38.53 nmol/L.

CONCLUSIONWe have obtained the ssDNA aptamers specific to the clinical isolates of highly cariogenic Streptococcus mutans strains.

Aptamers, Nucleotide ; genetics ; Cloning, Molecular ; DNA Primers ; Dental Caries ; microbiology ; Gene Library ; Humans ; Nucleic Acid Conformation ; SELEX Aptamer Technique ; Species Specificity ; Streptococcus mutans ; classification ; genetics ; isolation & purification