BACKGROUND: The biological behaviors of preadipocytes in adipose tissue of young animals have been closely linked to the onset and prevention and treatment of obesity. Observing mechanical oscillation effects on biomechanical behaviors of preadipocytes using biomechanical stimuli would provide more direct experimental evidence for treatment of simple obesity using manipulation and massage therapy.OBJECTIVE: Different frequencies of mechanical force stimulation were performed on preadipocytes from young rats cultured in vitro to observe the changes in cell viability, proliferation, and apoptosis.METHODS: Preadipocytes from SD young rats were in vitro cultured. Following identification, preadipocytes were dynamically, mechanically stimulated through the use of constant temperature oscillator. According to different treatment frequencies, three groups were set: 0 Hz (blank control), 1.5 Hz, and 3 Hz. A 30-minute oscillation, once every 12 hours, total 3 days, was performed in each group. Following dynamic mechanical stimulation, cell viability, proliferation, and apoptosis in young rats were observed. RESULTS AND CONCLUSION: In the initial stage of culture,cells exhibited the morphology similar to fibroblasts. After oil red O staining, red particles appeared in the cells, indicating that the young mouse cells cultured in vitro were preadipocytes. With stimulation of oscillation force, the viability and proliferation of preadipocytes were significantly inhibited (P < 0.05 or P < 0.01). There wasno significant difference in effects of mechanical oscillation on preadipocyte apoptosis between 0 Hz and 1.5 Hz, 3 Hz groups (P > 0.05). These findings indicate that thecell biological mechanism underlying preventing simple obesity in adolescents is to inhibit the viability and proliferation of preadipocytes

Objective To investigate the effect of the histological changes on rat dorsal root ganglia (DRG) after 125I seed brachytherapy.Methods Twelve adult male Sprague-Dawley rats ( 150-180g each) were randomly divided into 6 groups,125I seeds with different activities of 0 (Titanium shell),14.8,18.5,22.2,25.9 and 29.6 MBq were implanted to 6 groups of rats respectively and the behavioral changes of rats were observed.The rats were killed in different periods after implantation,the morphological changes in DRG and surrounding muscle tissue were observed with an Olympus BX51 optical microscope and then the irradiation doses were estimated.Results After 125I seed implantation,the movement function of rats was not affected and the weight of rats gained after 7 days.After the titanium shell implantation,very few mild swelling was induced in neuroganglion cells that still had clear nucleolus and normal cytoplasm.At 14 days after 18.5 MBq seed implantation,cell swelling was more serious and cell dehydrating,nuclear condensation and nuclear fragmentation appeared after 30 days.At 60 days after 29.6 MBq of seed implantation,nuclear dissolution and cytoplasmic shrinkage were induced in a large number of cells.In general,the severity of fibrosis was aggravated with the time post-irradiation and the dose in the muscles around the ganglion.Conclusions After 125I seed implantation,the injury degree of DRG tissue is dose-dependent,and the 125I seed irradiation would have analgesic effect on releasing intractable pain.

Objective To investigate the susceptibilities of HCMV clinical strains isolated to ganciclovir from the patients after hematopoietic stem cell transplantation.Methods Eight HCMV clinical isolates were isolated from the blood or the urine of hematopoietic stem cell transplantation recipients who had been treated with GCV.Tissue cell infection median dose(TCID50) were calculated by Reed-Muench method.Drug susceptibility was determined by MTT assay.Results TCID50 values of eight HCMV clinical strains were 10-4.12/0.1ml,10-4.29/0.1ml,10-4.3/0.1ml,10-4.4/0.1ml 10-4.42/0.1ml,10-4.5/0.1ml,10-4.52/0.1ml and 10-4.62/0.1ml respectively.50％ inhibitory concentration(IC50) to GCV of eight HCMV clinical strains were 0.638,1.438,0.965,0.698,0.482,1.167,1.519,1.511 mg/L respectively.Conclusion Our results suggest that resistant HCMV strains are not prevalent in Guangzhou.Continuous monitoring of HCMV is needed to understand the antiviral resistance status of the virus in patients after hematopoietic stem cell transplantation and guide its clinical management.

Objective To explore the mechanism underlying the rapid shortening of outflow tract and the formation of the right ventricle of the embryonic mouse heart .Methods Serial sections of embryonic mouse hearts from embryonic day 9 (E9) to E12(3 to 5 embryos for each stage)were stained with antibodies against α-sarcomeric actin (SCA), α-smooth muscle actin (SMA), GATA-4, myosin heavy chain (MHC), proliferating cell nuclear antigen (PCNA) or active caspase-3 (CAS-3).Results At E11, the aortic sac and the distal border of cardiac outflow tract had regressed towards the ventricle into the pericardial cavity , while GATA-4、SCA and SMA staining showed that precursors from the second heart field were differentiating into cardiomyocytes adding to the arterial pole of the heart to lengthen the outflow tract .The length of outflow tract rapidly shortened at E12.Before and during its shortening , no CAS-3 positive cell was detected in the entire outflow tract.During E10-12, the cardiomyocytes in the right ventricle and proximal outflow tract wall proliferated inward to form trabeculae, with some trabeculae extending into the ridges .Proximal extremities of the outflow tract ridges were gradually myocardialized remodeling into the trabeullar right ventricle wall .At E12, scattered SCA and SMA staining cells and SCA and SMA weak positive mesenchymal cell clusters , which were continuous with the outflow tract myocardium were detected in the mesenchymal proximal outflow tract ridges .These results suggested that the proximal outflow tract was remodeled into the right ventricle by trabecularization , during which mesenchymal ridges were trabecularlly myocardialized . Conclusion Ventricularization of the proximal outflow tract contributes to the trabecular right ventricle and resultes in the vapid shortening of outflow tract in the mouse embryonic heart .Cardiomyocyte appoptosis and transdifferentiation are found to play a more limited contribution during this process .

Objective To evaluate the effects of Ulinastatin (UTI) on the expressions of TNF-α,IL-6 and neurons apoptosis in cerebral cortex of rats after cardiopulmonary resuscitation (CPR).Methods Thirty-six healthy male adult Wistar rats were induced ventricular fibrillation untreated for 7 min and then received CPR.The animals were infused UTI 100 000 U/kg or phosphate-buffered solution (PBS) at once after ROSC.At 2,4 and 8 h after ROSC,cerebral cortex were removed to determine the mRNA expressions and levels of TNF-α protein and IL-6 protein,the translocation ratio of NF-κB p65 from cytoplasm to nucleus and the apoptotic neurons.Results The plasma levels of TNF-α (ng/mL) in animals of UTI group were (17.7 ± 1.4),(21.9 ± 2.1) and (17.1 ± 0.6) at 2,4 and 8 h after ROSC respectively,and significantly lower than those in PBS group at the given intervals.Mean while,the levels of IL-6 (ng/mL) were (208.9 ± 14.1),(281.5 ±25.9) and (251.8 ± 15.3) at 2,4 and 8 h after ROSC respectivèly in animals of UTI group,and lower than those in PBS group.The expressions of TNF-α mRNA and IL-6 mRNA and protein levels of TNF-α and IL-6 in UTI group were both lower than those in PBS group at given intervals,respectively.The translocation ratio of NF-κB p65 from plasma to nucleus in PBS group at each given interval after ROSC was significantly higher than that in UTI group.The number of viable neurons in cerebral cortex in UTI group was higher than that in PBS group,while the number apoptosis neurons was fewer in UTI group.Conclusions UTI attenuated the general inflammatory response after ROSC in rat,decreased the activation of NF-κB pathway,and subsequently attenuated the expression of TNF-α and IL-6,and finally decreased the neurons apoptosis.

Objective:To explore the effect of nursing goal execution concept-based intervention strategy in patients with bronchial asthma.Methods:From February 2018 to March 2021, 118 patients with bronchial asthma admitted to the First Affiliated Hospital of Zhengzhou University were selected by convenience sampling as the research object. The patients were randomly divided into control group and observation group with 59 cases each. The control group was given routine cognitive education, and the observation group received the intervention strategy based on nursing goal execution concept on the basis of the control group. The self-management ability, asthma control and quality of life of the two groups were compared after six months of follow-up.Results:Finally, 55 patients in the control group and 57 patients in the observation group completed the study. After intervention, the total score and each dimension score of self-management ability of the observation group were higher than those of the control group, the Asthma Control Test score was higher than that of the control group, and the total score and each dimension score of Asthma Quality of Life Questionnaire were higher than those of the control group, with statistical differences ( P<0.05) . Conclusions:The application of nursing goal execution concept-based intervention strategy in patients with bronchial asthma can improve their self-management ability, asthma control and quality of life, which is worthy of clinical application.

Objective To evaluate the effect of luteolin on the growth,migration and vasculogenic mimicry formation of a melanoma cell line B16.Methods In vitro cultured B16 melanoma cells were divided into 4 groups:low-,middle-and high-dose luteolin groups treated with 2.5,5,10 μmol/L luteolin respectively,and control group treated with 0.1％ dimethyl sulfoxide (DMSO).Scratch assay,Transwell invasion assay and vascular channel formation assay were performed to assess the migration,invasion of and vascular channel formation by melanoma cells.A model of subcutaneous transplanted B 16 melanoma was established in 12 C57 mice,which were randomly and equally divided into 4 groups:control group gavaged with ultrapure water,low-,middle-and high-dose luteolin groups gavaged with 10,20,40 mg/kg luteolin respectively every day.The above treatment for the tumor-bearing mice lasted till day 28,and then these mice were sacrificed.Meanwhile,the lung and tumor tissues of the mice were excised,and the growth,metastasis and vasculogenic mimicry of transplanted melanoma were observed.Immunofluorescence and immunohistochemical studies were performed to evaluate the effects of luteolin on the expression of vascular endothelial cadherin (VE-cadherin),vascular endothelial growth factor receptor 1 (VEGFR1),VEGFR2,matrix metalloproteinase-2 (MMP-2) and MMP-9 in the transplanted melanoma.Means were compared among several groups by using one-way analysis of variation or rank sum test.Results In vitro study showed that the relative scratch width at 48 hours significantly differed among the control group,low-,middle-and high-dose luteolin groups (0.47 ± 0.04,0.64 ± 0.04,0.73 ± 0.03,0.84 ± 0.04 respectively;F =34.51,P ＜ 0.001),and the migration ability of B16 cells was significantly lower in the low-,middle-and high-dose luteolin groups than in the control group (all P ＜ 0.05).At 24 hours,there were significant differences in the number of cells crossing the Transwell membrane among the control group,low-,middle-and high-dose luteolin groups (281.00 ± 8.79,169.00 ± 15.35,92.00 ± 14.79 and 57.00 ± 13.72 respectively;F =275.30,P ＜ 0.001),and the invasive ability was significantly lower in the low-,middle-and high-dose luteolin groups than in the control group (P ＜ 0.01).Meanwhile,the number of formed vascular channels also differed among the above 4 groups (20.00 ± 2.77,11.00 ± 1.28,7.00 ± 1.86 and 2.00 ± 1.32 respectively;F =48.61,P ＜ 0.001),and the number of vascular channels was significantly lower in the low-,middle-and high-dose luteolin groups than in the control group (all P ＜ 0.01).In vivo study showed that the tumor size significantly differed among the control group,low-,middle-and high-dose luteolin groups (5.10 ± 1.72,4.02 ± 2.13,2.98 ± 0.92,1.49 ± 1.13 cm3 respectively;F =28.76,P ＜ 0.001),and was significantly lower in the low-,middle-and high-dose luteolin groups than in the control group (t =3.86,7.11 and 13.06 respectively,all P ＜ 0.01).CD31-PAS double staining showed that the number of vasculogenic mimicry was significantly higher in the control group than in the low-,middle-and high-dose luteolin groups (all P ＜ 0.01).In vivo and in vitro studies both showed that the expression of vasculogenic mimicry-related markers in the cells or mouse tumor tissues was significantly lower in the high-dose luteolin group than in the control group (P ＜ 0.05).Conclusion Luteolin can effectively inhibit the growth,metastasis and vasculogenic mimicry formation of melanoma.

Objective To monitor human cytomegalovirus (HCMV) drug resistance in recipients of hematopoietic stem cell transplantation by phenotypic and genotypic methods. Methods HCMV clinical isolates was isolated from the urine of hematopoietic stem cell transplantation recipients treated with GCV. Tissue cell infection median dose (TCID50) of the isolates was calculated using Reed-Muench method, and their drug susceptibility was determined by plaque reduction assay. We amplified the UL97 DNA fragment of the virus by nested PCR followed by automated DNA sequencing. Results HCMV clinical strain isolated from the urine samples of the recipients using a human fibroblast cell line showed a TCID50 value of 10-4.618/0.1ml and a 50%inhibitory concentration (IC50) to GCV of 5.847 μmol/L, suggesting its sensitivity to GCV. Alignment with the AD169 DNA reference sequence identified 4 point mutations of the virus at 1509 (T-C), 1575 (C-T), 1794 (T-C), and 1815 (C-G), and only the last mutation resulted in one amino acid mutation to D605E. No gene mutation was found in relation to GCV resistance. Conclusions Phenotypic and genotypic assays were established to examine antiviral drug resistance of HCMV in recipients of hematopoietic stem cell transplantation. We did not find any drug resistance of the clinical HCMV isolate.

Objective To monitor human cytomegalovirus (HCMV) drug resistance in recipients of hematopoietic stem cell transplantation by phenotypic and genotypic methods. Methods HCMV clinical isolates was isolated from the urine of hematopoietic stem cell transplantation recipients treated with GCV. Tissue cell infection median dose (TCID50) of the isolates was calculated using Reed-Muench method, and their drug susceptibility was determined by plaque reduction assay. We amplified the UL97 DNA fragment of the virus by nested PCR followed by automated DNA sequencing. Results HCMV clinical strain isolated from the urine samples of the recipients using a human fibroblast cell line showed a TCID50 value of 10-4.618/0.1ml and a 50%inhibitory concentration (IC50) to GCV of 5.847 μmol/L, suggesting its sensitivity to GCV. Alignment with the AD169 DNA reference sequence identified 4 point mutations of the virus at 1509 (T-C), 1575 (C-T), 1794 (T-C), and 1815 (C-G), and only the last mutation resulted in one amino acid mutation to D605E. No gene mutation was found in relation to GCV resistance. Conclusions Phenotypic and genotypic assays were established to examine antiviral drug resistance of HCMV in recipients of hematopoietic stem cell transplantation. We did not find any drug resistance of the clinical HCMV isolate.

OBJECTIVETo monitor human cytomegalovirus (HCMV) drug resistance in recipients of hematopoietic stem cell transplantation by phenotypic and genotypic methods.

METHODSHCMV clinical isolates was isolated from the urine of hematopoietic stem cell transplantation recipients treated with GCV. Tissue cell infection median dose (TCID50) of the isolates was calculated using Reed-Muench method, and their drug susceptibility was determined by plaque reduction assay. We amplified the UL97 DNA fragment of the virus by nested PCR followed by automated DNA sequencing.

RESULTSHCMV clinical strain isolated from the urine samples of the recipients using a human fibroblast cell line showed a TCID50 value of 10(-4.618)/0.1 ml and a 50% inhibitory concentration (IC50) to GCV of 5.847 µmol/L, suggesting its sensitivity to GCV. Alignment with the AD169 DNA reference sequence identified 4 point mutations of the virus at 1509 (T-C), 1575 (C-T), 1794 (T-C), and 1815 (C-G), and only the last mutation resulted in one amino acid mutation to D605E. No gene mutation was found in relation to GCV resistance.

CONCLUSIONSPhenotypic and genotypic assays were established to examine antiviral drug resistance of HCMV in recipients of hematopoietic stem cell transplantation. We did not find any drug resistance of the clinical HCMV isolate.

Antiviral Agents ; pharmacology ; Cell Line ; Cytomegalovirus ; drug effects ; genetics ; isolation & purification ; Drug Resistance, Viral ; genetics ; Ganciclovir ; pharmacology ; Genes, Viral ; Genotype ; Hematopoietic Stem Cell Transplantation ; Humans ; Mutation ; Phosphotransferases (Alcohol Group Acceptor) ; genetics