[Effect of HCMV infection on immune reconstitution of CD8+T cells in children with allogeneic hematopoietic stem cell transplantation].

Objective: To investigate the risk factors for human cytomegalovirus infection after allogeneic hematopoietic stem cell transplantation in children and the impact of human cytomegalovirus infection on post-transplant immune reconstitution. Methods: A Retrospective Co-Hort study design was used to include 81 children treated with allo-HSCT from January 2020 to March 2022 at the Department of Hematology, Capital Institute of Pediatrics, Beijing, China, and followed up for 1 year. Real-time quantitative PCR was used to detect positive detection of HCMV in children after allo-HSCT, multifactorial logistic regression modeling was used to analyze the risk factors leading to HCMV infection, and generalized estimating equation modeling was used to analyze the effect of HCMV infection on the T-cells of the children who received allo-HSCT. Results: The age M(Q1, Q3) of 81 children was 5.1 years (10 months, 13.8 years), and 50 (61.7%) were male. By the endpoint of follow-up, a total of 50 HCMV-positive cases were detected, with an HCMV detection rate of 61.7%; The results of multifactorial logistic regression modeling showed that children with grade 2-4 aGVHD had a higher risk of HCMV infection compared with grade 0-1 after transplantation [OR (95%CI) value: 2.735 (1.027-7.286)]. The results of generalized estimating equation modeling analysis showed that the number of CD3+T cells in HCMV-positive children after transplantation was higher than that in the HCMV-negative group [RR (95%CI) value: 1.34 (1.008-1.795)]; the ratio of CD4+T/CD8+T cells was smaller than that in the HCMV-negative group [RR (95%CI) value: 0.377 (0.202-0.704)]; the number of CD8+T cells was higher than that in the HCMV-negative group [RR (95%CI) value: 1.435 (1.025-2.061)]; the number of effector memory CD8+T cells was higher than that in the HCMV-negative group [RR (95%CI) value: 1.877 (1.089-3.236)]. Conclusion: Acute graft-versus-host disease may be a risk factor for HCMV infection in children after allo-HSCT; post-transplant HCMV infection promotes proliferation of memory CD8+T-cell populations and affects immune cell reconstitution.

[Endemic situation of schistosomiasis in national surveillance sites of Zhejiang Province from 2015 to 2019].

OBJECTIVE: To analyze the endemic situation of schistosomiasis in national surveillance sites of Zhejiang Province, so as to provide the scientific evidence for formulating the new schistosomiasis surveillance program and consolidating schistosomiasis elimination achievements. METHODS: A total of 54 to 55 national surveillance sites and 6 risk monitoring sites were assigned in historical endemic regions of Zhejiang Province from 2015 to 2019, where the Schistosoma japonicum infections in humans, livestock, snails and wild feces were monitored. All data pertaining to the surveillance results were descriptively analyzed. RESULTS: There were 34 530 person-time local residents receiving serological screening for S. japonicum infections in national surveillance sites of Zhejiang Province from 2015 to 2019, and the seroprevalance of S. japonicum infections were 0.29% to 0.68%; however, no egg-positives were detected. During the period from 2015 to 2019, there were 62 086 person-time mobile populations receiving serological screening for S. japonicum infections in national surveillance sites of Zhejiang Province, and the seroprevalance of S. japonicum infections were 0.32% to 0.56%; additionally, there were 27 egg-positives identified, including 3 imported cases with schistosomiasis haematobia and one case with acute schistosomiasis japonica, indicating that the adjusted prevalence of Schistosoma infections were 0.01% to 0.07% among mobile populations. A total of 829 livestock were examined for S. japonicum infections, with no positives detected. There were 10.55 hm2 and 2.89 hm2 snail habitats found in national surveillance sites and risk monitoring sites of Zhejiang Province during the 5-year period, respectively; however, no S. japonicum infections were identified in snails. Moreover, a total of 305 wild feces were tested for S. japonicum infections, and no egg-positives were identified. CONCLUSIONS: The schistosomiasis elimination achievement have been consolidated in Zhejiang Province; however, the risk factors of re-emerging schistosomiasis have not been completely eliminated, such as residual snails and imported schistosomiasis cases. Further surveillance and control activities of snails and the source of S. japonicum infections should be reinforced.

CalliSpheres® drug-eluting beads (DEB) transarterial chemoembolization (TACE) is equally efficient and safe in liver cancer patients with different times of previous conventional TACE treatments: a result from CTILC study.

PURPOSE: To assess the efficacy and safety of drug-eluting beads transarterial chemoembolization (DEB-TACE) in liver cancer patients with different times of previous conventional transarterial chemoembolization (cTACE) treatments. METHODS: 367 liver cancer patients about to receive DEB-TACE treatment were enrolled in this prospective cohort study. All patients were divided into no previous cTACE group (NPC group), 1-2 times previous cTACE group (PC group) and triple or above previous cTACE group (TPC group) according to the times of previous cTACE treatments. RESULTS: There was no difference in complete response (CR) (P = 0.671) and objective response rate (ORR) (P = 0.062) among three groups. Additionally, no difference in overall survival (OS) among groups (P = 0.899) was found. As to liver function, most liver function indexes were deteriorative at 1 week after DEB-TACE operation, but returned to baseline at 1-3 months after DEB-TACE operation in all three groups, while percentage of abnormal total bile acid (TBA) patients was higher in TPC group than NPC and PC groups at 1-3 month post-DEB-TACE (P = 0.018). As for safety profiles, the incidence of pain during DEB-TACE operation was lower in TPC group compared to NPC and PC groups (P = 0.005), while no difference of other adverse events was found during and 1 month post-DEB-TACE treatment among three groups. CONCLUSION: DEB-TACE treatment was equally efficient and tolerated in liver cancer patients with different times of previous cTACE treatments.

Unique genetic profiles from cerebrospinal fluid cell-free DNA in leptomeningeal metastases of EGFR-mutant non-small-cell lung cancer: a new medium of liquid biopsy.

Background: Leptomeningeal metastases (LM) are more frequent in non-small-cell lung cancer (NSCLC) with epidermal growth factor receptor (EGFR) mutations. Due to limited access to leptomeningeal lesions, the purpose of this study was to explore the potential role of cerebrospinal fluid (CSF) as a source of liquid biopsy in patients with LM. Patients and methods: Primary tumor, CSF, and plasma in NSCLC with LM were tested by next-generation sequencing. In total, 45 patients with suspected LM underwent lumbar puncture, and those with EGFR mutations diagnosed with LM were enrolled. Results: A total of 28 patients were enrolled in this cohort; CSF and plasma were available in 26 patients, respectively. Driver genes were detected in 100% (26/26), 84.6% (22/26), and 73.1% (19/26) of samples comprising CSF cell-free DNA (cfDNA), CSF precipitates, and plasma, respectively; 92.3% (24/26) of patients had much higher allele fractions in CSF cfDNA than the other two media. Unique genetic profiles were captured in CSF cfDNA compared with those in plasma and primary tissue. Multiple copy number variations (CNVs) were mainly identified in CSF cfDNA, and MET copy number gain identified in 47.8% (11/23) of patients was the most frequent one, while other CNVs included ERBB2, KRAS, ALK, and MYC. Moreover, loss of heterozygosity (LOH) of TP53 was identified in 73.1% (19/26) CSF cfDNA, which was much higher than that in plasma (2/26, 7.7%; P < 0.001). There was a trend towards a higher frequency of concomitant resistance mutations in patients with TP53 LOH than those without (70.6% versus 33.3%; P = 0.162). EGFR T790M was identified in CSF cfDNA of 30.4% (7/23) of patients who experienced TKI progression. Conclusion: CSF cfDNA could reveal the unique genetic profiles of LM and should be considered as the most representative liquid biopsy medium for LM in EGFR-mutant NSCLC.

Screening key miRNAs for human hepatocellular carcinoma based on miRNA-mRNA functional synergistic network.

The safety of miRNAs has been proven and the prophylactic use of miRNA-based approaches may be foreseen for patients with hepatocellular carcinoma (HCC). However, the underlying regulatory mechanism of miRNAs in HCC has not been fully clarified. Using bioinformatic analyses, we compared data of miRNA and mRNA expression profiling of HCC from Gene Expression Omnibus (GEO) database, respectively. Differentially expressed miRNAs and differentially expressed genes (DEGs) were identified. Based on the miRTarBase predictions, the miRNA-dependent regulatory network was constructed. In total, comparative analysis of five mRNA datasets and two miRNA datasets led to 1449 DEGs and 17 differentially expressed miRNAs, respectively. Based on the predictions, a global miRNA-mRNA regulatory network was then constructed, which encompassed 451 miRNA target gene pairs whose expressions were inversely correlated. Three miRNAs (miR-641, miR-507 and miR-501-5p) were the most connected miRNAs that regulated a large number of genes, among which miR-641 and miR-507 were novel miRNAs altered in HCC. We suggested that miR-501-5p will represent a powerful therapeutic target for HCC. Moreover, four up-regulated miRNAs (miR-769-3p, miR-941, miR-362-3p and miR-16-1) and one down-regulated miRNA (miR-581) may be involved in HCC. Additionally, two targets of MAPK8 and SRPK2 were also detected in this study, whose roles in HCC will be notable. In conclusion, we developed an integrative approach to construct an interactive global network of miRNA-mRNA, which can contribute to refine miRNA target predictions for developing new therapeutic approaches.

Molecular characterization and expression pattern of dmrt1 in the immature Chinese sturgeon Acipenser sinensis.

In this study, the cDNA of dmrt1 gene from the Chinese sturgeon Acipenser sinensis was isolated and its expression pattern was characterized in different tissues of immature A. sinensis. By real-time quantitative PCR (qrtPCR) analysis, the A. sinensis dmrt1 mRNA was detected mainly in gonad and with a higher level in the testis than the ovary, especially in 3 and 4 year-old samples. This indicated that the dmrt1 expression exhibited gradual testis specificity with development. The subcellular localization analysis indicated that the Dmrt1 protein exists only in germ cells and not in somatic cells. These results suggest that A. sinensis dmrt1 might be a highly specific sex differentiation gene for testis development and spermatogenesis.

[Correlation of FibroTouch and FibroScan with the stage of primary biliary cirrhosis].

Objective: To investigate the diagnostic value of FibroTouch and FibroScan for the stage of primary biliary cirrhosis (PBC). Methods: A total of 66 PBC patients who visited our hospital from January 2014 to March 2016 were enrolled, and all the patients underwent liver biopsy and FibroTouch and FibroScan tests. Liver stiffness measurement (LSM) was used to assess fibrosis degree, and the receiver operating characteristic (ROC) curve was used to compare the cut-off values, sensitivities, and specificities of these two methods in determining fibrosis stage. The Spearman rank correlation test was used to investigate the correlation between FibroTouch and FibroScan values. Results: The correlation coefficients between FibroTouch or FibroScan values and fibrosis stage determined by liver biopsy were 0.904 and 0.880, respectively (both P < 0.01). The cut-off values of FibroTouch in the diagnosis of PBC with fibrosis stages of ≥S1, ≥S2, ≥S3, and ≥S4 were 6.25 kPa, 9.05 kPa, 11.75 kPa, and 18.95 kPa, respectively, with sensitivities of 89.7%, 94.7%, 80.0%, and 80.0% and specificities of 100%, 100%, 87.0%, and 100%, respectively; the cut-off values of FibroScan were 6.05 kPa, 8.85 kPa, 12.40 kPa, and 16.20 kPa, respectively, with sensitivities of 96.4%, 88.6%, 76.2%, and 100% and specificities of 77.8%, 100%, 86.4%, and 93.0%, respectively. There were no significant differences in the diagnostic performance between FibroTouch and FibroScan in determining fibrosis stage [≥S1 (P = 0.109), ≥S2 (P = 0.853), ≥S3 (P = 0.387), ≥S4 (P = 0.224)]. Conclusion: FibroTouch and FibroScan can be used as noninvasive diagnostic tools for the determination of fibrosis stage and the monitoring of disease progression in PBC patients and have good sensitivity and specificity.

Osteoblastic cell response on fluoridated hydroxyapatite coatings: the effect of magnesium incorporation.

Magnesium (Mg) ions were incorporated into fluoridated hydroxyapatite (HA) coating by the sol-gel dip-coating method. Mg in the coating was measured by x-ray photoelectron spectroscopy (XPS). The changes of calcium and magnesium concentrations were recorded to monitor the dissolution behavior of the coatings. In vitro cell responses were evaluated using MG63 cells in terms of cell morphology, proliferation and differentiation. The substitution of Mg and F ions into the HA crystal structure was confirmed by XPS. Only a limited amount of Mg can be incorporated into HA lattice. The dissolution test revealed that Mg incorporation increased the solubility of the coating in the tris-buffered saline solution. The highest solubility was achieved at x = 1.5 (Ca((10-x))Mg(x)(PO(4))(6) F(OH). In the cell culture test, well-spread cells were observed on all the coatings. Also, a significantly positive effect of Mg ions on cell proliferation and late differentiation was found at x = 1.5. Mg incorporation stimulates osteoblastic cell responses on fluoridated hydroxyapatite coatings.

Efficacy and safety of catnip (Nepeta cataria) as a novel filth fly repellent.

Catnip (Nepeta cataria) is known for its pseudo-narcotic effects on cats. Recently, it has been reported as an effective mosquito repellent against several Aedes and Culex species, both topically and spatially. Our laboratory bioassays showed that catnip essential oil (at a dosage of 20 mg) resulted in average repellency rates of 96% against stable flies, Stomoxys calcitrans (L.) and 79% against houseflies, Musca domestica (L.), respectively. This finding suggested that the application of repellent could be used as part of filth fly management. Further evaluations of catnip oil toxicity were conducted to provide a broad-spectrum safety profile of catnip oil use as a potential biting and nuisance insect repellent in urban settings. Acute oral, dermal, inhalation, primary dermal and eye irritation toxicity tests were performed. The acute oral LD(50) of catnip oil was found to be 3160 mg/kg body weight (BW) and 2710 mg/kg BW in female and male rats, respectively. The acute dermal LD50 was > 5000 mg/kg BW. The acute inhalation LD50 was observed to be > 10,000 mg/m3. Primary skin irritation tested on New Zealand white rabbits showed that catnip oil is a moderate irritant. Catnip oil was classified as practically non-irritating to the eye. In comparison with other U.S. Environmental Protection Agency-approved mosquito repellents (DEET, picaridin and p-menthane-3,8-diol), catnip oil can be considered as a relatively safe repellent, which may cause minor skin irritation.

Migration of retinal pigment epithelial cells in vitro modulated by monocyte chemotactic protein-1: enhancement and inhibition.

BACKGROUND: The migration of retinal pigment epithelial (RPE) cells is an initial step in the development of proliferative vitreoretinopathy (PVR). This in vitro study was carried out to investigate the effects of monocyte chemotactic protein-1 (MCP-1) on the migration and proliferation of RPE cells. METHODS: We used an in vitro wound healing model in which a small area of a confluent monolayer of human RPE (HRPE) cells was denuded with a razor blade. The cultures were subsequently incubated with MCP-1, IL-1beta, TNF-alpha, or combinations thereof. Neutralizing IgG1 of antihuman MCP-1, dexamethasone (DEX) or daunorubicin were also added to the cultures to test their inhibitory effects on migration of RPE cells. HRPE migration was measured as the number of cells that entered the denuded area. The effect of MCP-1 on proliferation of HRPE cells was examined by MTT assay. RESULTS: MCP-1 stimulated HRPE cell migration in a dose-dependent manner. IL-1beta or TNF-alpha slightly stimulated HRPE cell migration, but adding anti-MCP- IgG1 significantly reduced this effect. MCP-1-induced migration could be inhibited by DEX but not by daunorubicin. MCP-1 did not show a significant effect on HRPE cell proliferation. CONCLUSION: MCP-1 stimulates HRPE cell migration, suggesting that this chemokine regulates the development of PVR at the initial stage. The migration of HRPE cells induced by IL-1beta and TNF-alpha may be associated with the MCP-1 that HRPE cells secretes under the stimulation of these two cytokines. The knowledge that MCP-1-induced migration of HRPE cells is inhibited by DEX may be useful in devising an effective treatment for PVR.

[Increased cellular proliferation in BEL-7402 hepatoma cells transfected by human hepatic stimulator substance gene].

A hepatotropic factor originally identified in weanling or regenerating rat livers, known as hepatic stimulator substance (HSS), is characterized by promoting markedly liver regeneration after partial hepatectomy. Although HSS gene and the gene product have been described recently, the cellular mechanism of HSS and its genetic function remain obscure. In our previous studies, human HSS (hHSS) was demonstrated to promote the growth of hepatoma cells and phosphorylate the mitogen-activated protein kinase (MAPK). In this study, we reported the growth features of human hepatoma cells response to hHSS gene transfection. The hHSS eukaryotic vector was transfected to BEL-7402 hepatoma cells and the expression of hHSS was analyzed with Northern and Southern blot. The results showed that the HSS recombinant construct was functionally expressed in the target cells as analyzed with Northern blot and reverse transcriptase polymerase chain reaction (RT-PCR). The growth potential of HSS transfected hepatoma cells was markedly enhanced as compared with the non transfected cells. The S-phase of the hHSS-transfected cells increased by 61.5% than that of the non-transfected cells as shown by flow cytometry. Moreover, MAPK phosphorylation, one of the most important mitogenic indexes of growth signal cascade, was profoundly activated at sites of Thr202/Tyr204 due to HSS gene transfer. Based on these results, it is concluded that the introduction of HSS gene into hepatoma cells might be able to stimulate directly the cellular growth in vitro, allowing a possibility of reevaluation of HSS mechanism in intact cells.

Transmission of afferent information from urinary bladder, urethra and perineum to periaqueductal gray of cat.

The micturition reflex pathway is a supraspinal pathway. Anatomical tracing evidence is compatible with an involvement of the periaqueductal gray (PAG) in the ascending limb of this reflex. We tested the involvement of the PAG in receiving urinary tract- or perineum-related information and attempted to characterize this ascending path in terms of what type of information is being conveyed. Electrical stimulation of the pelvic nerves, which carry afferent information from the urinary bladder, evoked maximum field potentials in the caudal third of the PAG, primarily in the dorsal part of the lateral PAG and in the ventrolateral PAG. Since the regions activated by pelvic nerve stimulation differed from those activated by stimulation of the sensory pudendal or superficial perineal nerves, it is possible that specific pathways for different nerve inputs to the PAG exist. Sacral spinal cord neurons ascending to the PAG were identified by antidromic activation and then tested for inputs from pelvic, sensory pudendal or superficial perineal nerves. Of 18 units identified, only five received inputs from any of the peripheral nerves tested and only two projecting neurons received a pelvic nerve input. Thus the PAG may receive inputs from bladder and perineum, but the small proportion of cells with direct projections to the PAG receiving inputs from our test nerves implies that the major part of this pathway is not directly related to lower urinary tract function.

Sacral spinal cord neurons responsive to bladder pelvic and perineal inputs in cats.

In chloralose-anesthetized or decerebrate male cats, 70% of 73 sacral spinal cord neurons activated from the bladder branch of the pelvic nerve also received excitatory inputs from urethra and/or perineal cutaneous nerves (sensory pudendal in 55% and superficial perineal in 84% of neurons). Only 29% of these neurons were excited by the hindlimb skin and muscle nerves tested. The pelvic nerve-responsive neurons received monosynaptic urethral/perineal input in 25% of cases and required temporal summation of this input in 47% of cases. Of 211 neurons responding to superficial perineal nerve stimulation, 101 were not excited by the other nerves tested. Neurons activated by superficial perineal nerve stimulation were found predominantly in S2. It is likely that the superficial perineal nerve represents an important pathway whereby perineal stimulation influences bladder function.

Serotonergic modulation of spinal ascending activity and sacral reflex activity evoked by pelvic nerve stimulation in cats.

Serotonin (5-HT) may be inhibitory to micturition at a spinal level. A potential mechanism of action for serotonergic inhibition of bladder function is a depression of the ascending limb of the supraspinal reflex mediating micturition. Ascending activity evoked by pelvic nerve stimulation was recorded in the thoracic spinal cord of anesthetized cats. For comparison, spinal reflex activity evoked by pelvic nerve stimulation was recorded on the pudendal nerve. The effects of intrathecal administration of serotonergic agents were examined to determine whether spinal and supraspinal responses to bladder afferent activation were modulated by 5-HT. Methysergide (60 nmol), a non-selective serotonergic antagonist, increased ascending activity by 61+/-7% and depressed spinal reflex activity by 38+/-6%. Zatosetron (10 nmol), a 5-HT3 antagonist had a similar effect on both activities (increased by 93+/-24% and decreased by 77+/-7%, respectively). The effect on ascending activity of blocking 5-HT3 receptors was also confirmed with ICS 205930 and MDL 72222. 2-Methyl-5-HT (800 nmol), a 5-HT3 agonist, depressed ascending activity to 46+/-9% of control, but enhanced spinal reflex activity by 73+/-92%. These results demonstrate that stimulation of 5-HT3 and methysergide-sensitive 5-HT receptors can inhibit ascending activity and facilitate spinal reflex activity elicited by activation of bladder afferents. It is suggested that descending serotonergic pathways may participate in the spinal coordination of urinary continence.

Medullary neurons with projections to lamina X of the rat as demonstrated by retrograde labeling after HRP microelectrophoresis.

Brainstem neurons were retrogradely labeled with microelectrophoresis of HRP or WGA-HRP into lamina X of the cervical or lumbar cord of rats. The results reveal that lamina X of the lumbar cord receives bulbar projections originating mainly within the nucleus raphe magnus and the nucleus reticularis paragigantocellularis (including the medial or alpha-ventral part and lateral part) and that lamina X of the cervical cord receives projections from similar but more extensive regions in the lower brainstem. These findings provide a neuroanatomical substrate for medullary descending modulation of nociceptive transmission in lamina X.

Inhibition of nociceptive responses of lumbar dorsal horn neurones by remote noxious afferent stimulation in the cat.

In cats anaesthetized with nitrous oxide and sodium pentobarbital, multireceptive lumbar dorsal horn neurones excited by controlled noxious radiant heating of glabrous hind paw skin were recorded by extracellular microelectrodes. These noxious heat responses were inhibited by concomitant noxious stimulation of the ipsilateral forepaw or pinna, or repetitive electrical stimulation of the ipsilateral forelimb deep radial nerve. Similar extents of inhibition were produced by noxious peripheral stimulation and by deep radial nerve stimulation in repetitive trains at intensities sufficient to excite small myelinated fibres or unmyelinated fibres. A greater inhibitory effect was produced by continuous repetitive high-intensity stimulation of the deep radial nerve. With a constant frequency (5 Hz) of continuous deep radial nerve stimulation, graded increases in stimulation intensity revealed the threshold for inhibition in the small myelinated fibre range, and an additional increment of the inhibitory effect when unmyelinated fibres were also activated. When suprathreshold for unmyelinated fibres, the efficacy of continuous deep radial nerve stimulation increased with graded increases in stimulation frequency, with a threshold frequency for inhibition between 0.5 and 1 Hz and maximal effect at 5 Hz. Two nociceptive-specific neurones studied were also inhibited by deep radial nerve stimulation. The results indicate that 'diffuse noxious inhibitory controls' (DNIC) occur in the cat and can be activated by remote electrical or natural noxious stimulation.

Localization of central structures involved in descending inhibitory effect of acupuncture on viscero-somatic reflex discharges.

The pain reaction provoked by visceral traction has been one of the main problems to be solved in acupuncture anesthesia. By means of recording the viscero-somatic reflex activity in the animals, we have found that this reflex activity can be inhibited by acupuncture, and that the inhibitory effect originates from the supraspinal structures. The central structures involved in the inhibitory effect of acupuncture have been identified in a series of transection and lesion experiments. In decerebrated animals the inhibitory effect during acupuncture was preserved, but the after-inhibition of acupuncture disappeared; transection at the lower medulla as well as the cervical cord abolished near completely the whole inhibitory effect of acupuncture; a median lesion in the medulla including the nucleus raphe maguns resulted in a significant decrease of the inhibitory effect during acupuncture and a complete disappearance of the after-inhibition of acupuncture. The available evidence indicates that the median region of the medulla is involved in the elaboration of the inhibitory effect of acupuncture on the viscero-somatic reflex. A suggestion is thus offered that these supraspinal structures may be the essential link of the centre which is activated by acupuncture and involved in the descending inhibitory influence on the pain reaction of visceral stimulation.