The megakaryocyte and platelet lineage may be one of the major sites of human cytomegalovirus (HCMV) infection. However, whether HCMV aggravates apoptosis in normal megakaryocytes was not well investigated. Megakaryocytic cell line CHRF-288-11 and HCMV AD 169 strain were co-cultured in this study. PCR was used to detect the direct infection of the cells by HCMV IEA expression. The apoptotic cells were analyzed by morphologic observation, DNA ladder formation, annexin V/PI and PI assay with flow cytometry. The results showed that HCMV significantly inhibited the growth of CHRF cells in three different concentrations of viral infection groups (10(-3), 10(-2), 10(-1)). The viability levels in each infection groups were 77%, 73% and 68% respectively after incubation for 7 days, compared with 98% in the control group. Using annexin V/PI with flow cytometry, it was shown that the percentages of apoptotic cells viral infection in groups (10(-3), 10(-2), 10(-1)) were (21.3 +/- 2.49)%, (25.8 +/- 3.65)% and (31.4 +/- 3.91)% at 7 days after infection, while the control was (3.68 +/- 1.47)%. The apoptotic cells were further confirmed by morphologic observation and DNA ladder formation. Furthermore, PCR detection also showed the direct infection by identification of HCMV IEA expression in CHRF cells. This study suggested that HCMV could directly infect megakaryocytes and aggravated apoptosis in HCMV-infected megakaryocytes.
Apoptosis ; Cell Survival ; Cells, Cultured ; Cytomegalovirus ; pathogenicity ; DNA, Viral ; analysis ; Humans ; Megakaryocytes ; cytology ; virology ; Polymerase Chain Reaction

Objective:To improve the genetic stability of HBV gene in transgenic mice.Methods:HBV transgenic mice were bred by backcross and double cross.The HBV gene expression and replication were studied with real-time PCR,ELISA and chemiluminescence.Results:The HBV transgenic mice have stably bred to 23rd generation.The serum HBsAg level is 4122.31?2044.74IU/ml;The rate of HBV transgenic mice whose serum HBV DNA reach 104~106copies/ml was 93.93%.The HBV replication and expression were improved markedly.There is no difference between male and female mice about serum HBsAg level.Conclusion:After breeding the HBV gene was expressed stably with high-level in transgenic mice.

Metabolome has become an important part of Systems Biology, and a large set of data has already gained by applying the methods of metabolome. How to deal with the data and how to combine data of metabolome with data of other omics are problems that can not be ignored. An Enzyme Amount Multiple Factor was imported into the enzyme kinetic equation. When the enzyme amount in the system changed, in silico model, it means to alter the Enzyme Amount Multiple Factor. In order to observe ethanol concentration response to enzyme amount changes in S. cerevisiae glycolysis pathway model, enzyme amount was separately set at high and low level, the corresponding Enzyme Amount Multiple Factor value was 10 and 0.1, relatively. Based on the result of simulation, twelve enzymes in pathway were separated into two classes, class I and class II by cluster analysis. The four enzymes belonging to class I, ADH, HK, PFK and PDC, all catalyze irreversible reactions. The six out of eight enzymes belonging to class II, ALD, GAPDH, GlcTrans, lpPEP, PGI and TIM, catalyze reversible reactions. The other two enzymes belonging to class II, lpGlyc and PK, catalyze irreversible reactions. Based on this method, data of metabolome and proteomics are easily integrated to accomplish relatively overall analysis of system properties.
Algorithms ; Cluster Analysis ; Computer Simulation ; Enzymes ; classification ; metabolism ; Ethanol ; metabolism ; Fungal Proteins ; metabolism ; Glycolysis ; Metabolic Networks and Pathways ; Metabolomics ; methods ; Models, Biological ; Saccharomyces cerevisiae ; metabolism ; Systems Biology ; methods

OBJECTIVETo study the immunoregulatory effect of hepatitis B virus (HBV) e antigen (HBeAg) on peripheral blood monocytes (PBMCs).

METHODSPBMCs were isolated from patients with chronic hepatitis B (CHB; both HBeAg- and HBeAg+) and healthy controls, and cultured with recombinant HBeAg. The HBeAg-induced changes in expression of PD-1/PD-L1 were measured by flow cytometry of the cells and in secreted cytokines were measured by enzyme-linked immunosorbent assay of the supernatants. Comparisons between two groups were made by the independent-samples t-test; the relationship between PD-1/B7-H1 level and HBV DNA copy number was evaluated by Spearman's correlation analysis.

RESULTSExposure to HBeAg led to a significant decrease in CD3+CD4+ T lymphocyte-specific expression of IFNa for both the CHB patients' and healthy controls' samples (t = 2.382 and -4.190 respectively, P less than 0.01). For the HBeAg- CHB patients' and healthy controls' samples, the HBeAg exposure led to increased levels of secreted cytokines IL-6, IL-10 and TNFa (t = 2.504, 3.583 and 4.324, P less than 0.01 and t = 3.542, 6.246 and 5.273, P less than 0.01 respectively) and of CD14+ PBMC-specific expression of PD-L1 (t = 4.815 and 3.454, P less than 0.05 respectively). Compared to the HBeAg-negative CHB patients' and healthy controls' samples, the HBeAg+ CHB patients' samples had significantly lower CD3+CD4+ T cell-specific expression of IFNa (t = -3.177 and -4.541, P less than 0.01 respectively), but significantly higher levels of secreted IL-4 (t = 3.382 and 4.393, P less than 0.01 respectively), of CD3+ T cells-specific expression of PD-1/PD-L1 (t = 4.755, 2.942 and 4.518, 4.595, P less than 0.01 respectively), and of CD14+ T cells-specific expression of PD-L1 (t = 5.092 and 5.473, P less than 0.01 respectively). The CD3+ T cells-specific expression of PD-L1 was significantly higher in the samples from HBeAg- CHB patients than from the healthy controls (t = 3.214, P less than 0.01).

CONCLUSIONHBeAg was able to down-regulate the production of Th1-type cytokines (IFNgamma), and up-regulate the secretion of Th2-type cytokines (IL-6, IL-10) and the expression of PD-1/PD-L1on monocytes. These changes are conducive to the formation of immune tolerance to HBV. Therefore, HBeAg may play an important role in immune tolerance to chronic HBV infection.

Adult ; Case-Control Studies ; Cells, Cultured ; Female ; Hepatitis B e Antigens ; genetics ; immunology ; Hepatitis B, Chronic ; blood ; immunology ; Humans ; Interferon-gamma ; immunology ; Interleukin-10 ; immunology ; Interleukin-6 ; immunology ; Leukocytes, Mononuclear ; immunology ; metabolism ; Male ; Middle Aged ; Recombinant Proteins ; immunology ; Th1 Cells ; immunology ; Th1-Th2 Balance ; Th2 Cells ; immunology

OBJECTIVETo confirm alternative methods for Mandarin hearing in noise test (MHINT) to get the performance-intensity (PI) function, and to set up right test rules for scoring sentence intelligibility for subjects.

METHODSSentence speech reception thresholds were obtained for 30 normally hearing subjects with the MHINT test using several adaptive rules based on either character or word segmentation of the sentences. Each adaptive rule was intended to measure the threshold at a different point on the PI function. By also measuring sentence intelligibility at each threshold, the accuracy with which the PI function was estimated could be evaluated for each type of segmentation. The results of different segmentation were compared.

RESULTSThere was no significant difference in the reception threshold of sentences (RTS) between the group which used character segmentation and the group which used word segmentation (P > 0.05); and also no significant difference in their speech recognition score (SRS, P > 0.05). When the same subject used both character segmentation and word segmentation to do the test, there was no significant difference in their RTS between character segmentation and word segmentation (P > 0.05); and also no significant difference in their SRS (P > 0.05).

CONCLUSIONSThis method could be used not only with normally hearing individuals and people with hearing loss to evaluate their sentence intelligibility, but also with people who had aided equipment.

Acoustic Stimulation ; Adolescent ; Adult ; Audiometry, Speech ; methods ; Female ; Humans ; Language ; Male ; Speech Intelligibility ; Speech Perception ; Young Adult

Objective To design a punching and positioning device for training Da Vinci surgical robots to improve the efficiency and accuracy of operation training.Methods The punching and positioning device mainly consisted of an abdominal girth,a connecting plate,a punching plate,a positioning disk and a punching cartridge.The abdominal girth had a long-strip shape and was made of stretch nylon elastic band;the connecting plate had a rectangular shape with its length direction perpendicular to the abdominal girth;the punching plate also had a rectangular shape and was made of hard silicone;the positioning plate was made of hard silicone and privided with a plated through hole;the punching cartridge were equipped with anti-fog components composed of heating wires and support strips,which had scale bars inside.Ten physicians were randomly divided into a control group and an experimental group,with 5 ones in each group.The two groups carried out positioning and punching respectively,with the traditional method for the control group and the device designed for the experimental group,which were compared in terms of the time consumption,ratio for lens fogging and number of collisions with the robotic arm.Results The experimental group had the time consumption and ratio for lens fogging lower significantly when compared with the control group(P＜0.05),while had no obvious difference in number of collisions with the robotic arm(P＞0.05),and the requirements of the training specifications were met.Conclusion The punching and positioning device has high efficiency and accuracy,and thus is worthy promoting for the operation training of surgical robots.[Chinese Medical Equipment Journal,2024,45(7):35-39]

ObjectivesThis study aims to use accelerometer to objectively evaluate daily physical activity （PA） and steps of single pregnant women， and preliminarily explore the threshold of daily steps in line with the recommended PA during pregnancy， so as to provide PA guidance for pregnant women. MethodsWomen with singleton pregnancies who had prenatal examination in the the First Affiliated Hospital of Sun Yat-sen University from April 2018 to September 2019 continuously wore an accelerometer for 7 days for three times to measure daily PA. A total of 178 cases were included in this study with the average age 30.0 ± 3.0 years. Data were downloaded by supporting software， and the relationship between daily steps and PA （total PA， light PA and moderate PA） were analyzed by linear correlation analysis. Linear regression analysis and receiver operator characteristic curve （ROC） analysis were both used to explore the threshold steps corresponding to the physical activity recommendation. ResultsThere was no difference in daily total PA， light PA and sedentary behavior time during the three trimesters （P > 0.05）. But the moderate PA decreased with the pregnancy stages increased （25.1±16.1 vs. 23.6±16.5 vs. 21.5±16.1，min/d， P = 0.001）. There was a positive correlation between total PA and daily steps during the three trimesters （The correlation coefficients r of the early pregnancy， mid pregnancy and late pregnancy were 0.87， 0.83 and 0.86， respectively， P < 0.001）； and there was a positive correlation with light PA （r of the three trimesters were 0.79， 0.72 and 0.77， respectively， P < 0.001）. The relationship between moderate PA and daily steps during the three trimesters also had similar results （r of the three trimesters were 0.62， 0.53 and 0.55， respectively， P < 0.001）. The thresholds of daily steps to reach the recommended PA during pregnancy in the early， mid and late pregnancy were 10 925 steps， 11 372 steps and 11 170 steps， respectively. According to ROC analysis， the thresholds of daily steps in the three trimesters were 10 856 steps， 10 480 steps and 9 819 steps， respectively， and the areas under the curve were 0.81， 0.73 and 0.74， respectively （P < 0.001）. ConclusionsThe daily physical activity is mainly walking during pregnancy. Based on accelerometer evaluation， daily walking 9 800 to 11 500 steps could reach the physical activity recommendation during pregnancy.

Objective To explore the electrophysiological results and rehabilitation outcome of two prelingually deafened pediatric cochlear implant patients with auditory neuropathy.Methods Preoperative audiological evaluation,intra-postoperative electrically evoked auditory brainstem response(EABR)and neural response telememetry(NRT)record for the two cases were conducted in Beijing Tongren Hospital.A one year follow-up was performed.Data collected before and at 6,12-month intervals after implantation were compared with that from control pediatric cochlear implant patients matched for the same duration of implant use as this two cases. Results The two children implanted had not had any postoperative medical or cochlear implant device complications. Intraoperative EABR and NRT were elicited in case 1 with unrepeatable waveforms. After 12 months of training,Case 1 had shown significant improvements in sound detection,speecn perception abilities and communication skills,which was better than the control group.and the eJectrophysiological results became normal.Case 2 had also benefited from cochlear implantation, even though no recognizable NRT was found until he returned 12 month after the operation.Conclusions The desynchronization of auditory path had been changed after the electrical stimulation ongoing 12 months for children with auditory neuropathy.The two children had not had any complications postoperatively,and eacn child had shown improved listening and communication skills. Cochlear implantation COUld help patients witn auditory neuropathy to improve their communication skill and go back to the main stream.

BACKGROUND: Enhanced recovery after surgery (ERAS) protocols are a series of perioperative care to optimize preoperative preparation, prevent postoperative complications, minimize stress, and speed up recovery. This study aimed to assess the impact of ERAS protocols for functional endoscopic sinus surgery (FESS) in patients with chronic rhinosinusitis with nasal polyps (CRSwNP). METHODS: One hundred and two patients with CRSwNP undergoing FESS were randomly divided into the ERAS group and the control group. The outcomes of the Self-Rating Anxiety Scale (SAS), Visual Analogue Scale (VAS), Medical Outcomes Study Sleep Scale (MOS-SS) and Kolcaba Comfort Scale Questionnaire (GCQ) were determined in both groups. The serum levels of C-reactive protein (CRP) were compared preoperatively and 24 hours postoperatively. RESULTS: The ERAS group had a significantly better SAS scores than did the control group (28 [24, 35] vs. 43 [42, 47], Z = 5.968, P < 0.001). The rhinalgia and headache scores at 2, 24 and 48 hours postoperatively were lower in the ERAS group than that in the control group (all P < 0.001). The outcomes of the MOS-SS (43 [42, 39] vs. 28 [22, 35], Z = 7.071, P < 0.001) and GCQ (76 [68, 87] vs. 64 [50, 75], Z = 4.806, P < 0.001) were significantly different between the two groups. No significant difference was found in the preoperative CRP levels between the two groups (1.3 [0.6, 2.8] vs. 0.5 [0.5, 1.2], Z = 3.049, P > 0.05); However, the CRP level in 24 hours postoperatively was significantly lower in the ERAS group than that in the control group (2.5 [1.4, 3.9] vs. 6.6 [3.8, 9.0], Z = 5.027, P < 0.001). The incidence rates of complications, such as nausea/emesis (χ = 0.343, P > 0.05), hemorrhage, aspiration and tumble, were not increased in the ERAS group compared with those in the control group. The ERAS group had a significantly shorter length of hospital stay (5 [4, 5] days vs. 8 [8,9] days, Z = 8.939, P < 0.001) and hospitalization expenses ($ 2670 [2375, 2740] vs. $3129 [3116, 3456], Z = 8.514, P < 0.001). CONCLUSIONS: ERAS protocols might optimize FESS for patients with CRSwNP by reducing psychological and physical stress, shortening the length of hospital stay and lowering hospitalization expenses without increasing postoperative complications. TRIAL REGISTRATION Chinese Clinical Trial Registry, No. ChiCTR1800015791; http://www.chictr.org.cn/showproj.aspx?proj=26872.
Adolescent ; Adult ; C-Reactive Protein ; metabolism ; Chronic Disease ; Female ; Humans ; Length of Stay ; Male ; Middle Aged ; Nasal Polyps ; metabolism ; surgery ; Perioperative Care ; Postoperative Complications ; Postoperative Period ; Sinusitis ; metabolism ; surgery ; Surveys and Questionnaires ; Transanal Endoscopic Surgery ; methods ; Young Adult

Two cardenolide glycosides, corotoxigenin 3-O-[β-D-glucopyranosyl-(1→4)-6-deoxy-β-D-glucopyranoside] (1) and coroglaucigenin 3-O-[β-D-glucopyranosyl-(1→4)-6-deoxy-β-D-glucopyranoside] (2), were isolated from the seed fairs of Asclepias curassavica. The structures of 1-2 were determined based on the combination of the analysis of their MS, NMR spectroscopic data and acid hydrolysis. The inhibitory effects of compounds 1 and 2 on human colorectal carcinoma cells (HCT116), non-small cell lung carcinoma cells (A549) and hepatic cancer cells (SMMC-7721) were evaluated. The results showed that both compounds 1 and 2 significantly inhibited the viability, proliferation, and migration of A549, HCT116 and SMMC-7721 cells, suggesting that compounds 1 and 2 can be applied in the treatment of lung, colon and liver cancers in clinical practice. This study may not only provide a scientific basis for clarifying the active ingredients in A. curassavica, but also help to understand its antitumor activity, which can promote the application of A. curassavica in clinical treatment of various cancers.
Antineoplastic Agents/pharmacology* ; Asclepias/chemistry* ; Cardenolides/pharmacology* ; Glycosides/pharmacology* ; Humans ; Seeds