[Methylthioadenosine phosphorylase and p16 as surrogate diagnostic markers for CDKN2A homozygous deletion in brain tumors].

Objective: To examine whether immunohistochemistry of methylthioadenosine phosphorylase (MTAP) and p16 could be used to predict the CDKN2A status in various brain tumors. Methods: A total of 118 cases of IDH-mutant astrocytomas, 16 IDH-wildtype glioblastoma, 17 polymorphic xanthoastrocytoma (PXA) and 20 meningiomas diagnosed at Xuanwu Hospital, Capital Medical University, Beijing, China from November 2017 to October 2023 were collected and analyzed. The CDKN2A status was detected by using fluorescence in situ hybridization or next-generation sequencing. Expression of MTAP and p16 proteins was detected with immunohistochemistry. The association of loss of MTAP/p16 expression with CDKN2A homozygous/heterozygous deletion was examined. Results: Among the 118 cases of IDH-mutant astrocytoma, 13 cases showed homozygous deletion of CDKN2A. All of them had no expression of MTAP while 9 cases had no expression of p16. Among the 16 cases of IDH wild-type glioblastoma, 6 cases showed homozygous deletion of CDKN2A. All 6 cases had no expression of MTAP, while 3 of these cases had no expression of p16 expression. Among the 17 PXA cases, 4 cases showed homozygous deletion of CDKN2A, and the expression of MTAP and p16 was also absent in these 4 cases. Among the 20 cases of meningiomas, 4 cases showed homozygous deletion of CDKN2A. Their expression of MTAP and p16 was also absent. Among the four types of brain tumors, MTAP was significantly correlated with CDKN2A homozygous deletion (P<0.05), with a sensitivity of 100%. However, it was only significantly correlated with the loss of heterozygosity (LOH) of CDKN2A in astrocytomas (P<0.001). P16 was associated with CDKN2A homozygous deletion in IDH-mutant astrocytoma and PXA (P<0.001), but not with the LOH of CDKN2A. Its sensitivity and specificity were lower than that of MTAP. Conclusions: MTAP could serve as a predictive surrogate for CDKN2A homozygous deletion in adult IDH-mutant astrocytoma, PXA, adult IDH-wildtype glioblastoma and meningioma. However, p16 could only be used in the first two tumor types, and its specificity and sensitivity are lower than that of MTAP.

[Observation of visual quality after implantation of a rotational asymmetric refractive intraocular lens with a low addition region using the micromonovision design].

Objective: To investigate the effects of the application of a low addition refractive multifocal intraocular lens (IOL) using the micromonovision design in the non-dominant eye with different degrees of preset myopia on the visual acuity, visual function and visual quality after bilateral cataract surgery. Methods: In this randomized controlled trial, patients who were proposed to undergo bilateral phacoemulsification combined with rotational asymmetric refractive IOL (MF15 IOL) implantation at the First Affiliated Hospital of Zhengzhou University between September 2020 and August 2022 were included. All patients were divided into three groups using the random number method. The target refraction of the IOL in the dominant eye was 0.00 D. Non-dominant eyes were given different preoperative IOL reserve refractions, with the reserved near additional degree>0.20 D and≤0.40 D as the low addition,>0.40 D and≤0.60 D as the medium addition, and>0.60 D and≤0.80 D as the high addition. We compared uncorrected distance visual acuity (UDVA), uncorrected intermediate visual acuity (UIVA) and uncorrected near visual acuity (UNVA) of monocular and binocular eyes at 1 day, 1 month and 3 months postoperatively in the 3 groups of patients. Furthermore, the contrast sensitivity, stereopsis, defocus curves and visual quality questionnaire results of binocular eyes were compared at 3 months postoperatively. The statistical methods mainly used were chi-square test, two-factor repeated measures ANOVA, one-way ANOVA, LSD test, Kruskal-Wallis test, and paired t-test. Results: A total of 110 patients (220 eyes) were enrolled in the study, including 48 males and 62 females, with an average age of (59.74±9.38) years. There were 40 patients (80 eyes) in the low additional degree group, 37 patients (74 eyes) in the medium additional degree group, and 33 patients (66 eyes) in the high additional degree group. The differences in distance, intermediate and near visual acuity of the dominant eyes among the three groups were not statistically significant at different measurement timepoints postoperatively (P>0.05). The differences in intermediate and near visual acuity of the non-dominant eyes were also not statistically significant (P>0.05) among the three groups. In contrast, at 3 months, the UDVA of the non-dominant eyes in the low additional degree group (0.04±0.06) and medium additional degree group (0.04±0.07) was significantly higher than that in the high additional degree group (0.08±0.09) (F=4.776, P=0.011, bias η2=0.086). There was no statistically significant difference in binocular uncorrected distance, intermediate and near visual acuity among the three groups at different postoperative timepoints (P>0.05). The binocular UDVA, UIVA and UNVA (logMAR visual acuity) at 3 months postoperatively were -0.04±0.04, 0.03±0.08, 0.10±0.13 in the low addition group, -0.01±0.05, -0.02±0.06, 0.09±0.10 in the medium addition group, and 0.02±0.07, 0.01±0.09, 0.16±0.11 in the high addition group. At 3 months postoperatively, the binocular contrast sensitivity of the low additional degree group was significantly higher than that of the high additional degree group (P<0.05), except that there was no significant difference at the spatial frequency of 6 cycles per degree in the absence of glare (P>0.05). The binocular contrast sensitivity of the medium additional degree group was significantly higher than that of the high additional degree group at the spatial frequencies of 6 and 18 cycles per degree in the glare condition (P<0.05). The difference in the binocular contrast sensitivity between the low and medium additional degree groups did not reach statistical significance (P>0.05). The peak of the binocular defocus curve in the three groups was significantly wider than that in the monocular eyes, and the decline trend was more gentle, with no trough in the middle, and the visual acuity could be maintained above 0.2 (logMAR visual acuity) in the 0.00 D to -3.00 D defocus range. There was no significant difference in the postoperative near stereopsis results among the three groups (P>0.05), with the percentage of near stereopsis sharpness≤60″ reaching 90.00% (36/40), 89.19% (33/37) and 78.79% (26/33), respectively. The proportions of VF-14 scores≥90 in the postoperative questionnaire were 90% (36/40), 91.89% (34/37) and 81.82% (27/33) for the low, medium and high additional degree groups, respectively. The differences in the probability of photic phenomena and spectacles-independent rate were not statistically significant (P>0.05). Conclusion: The use of micromonovision design for bilateral implantation of a rotational asymmetric refractive MF15 IOL, with the non-dominant eye reserved for different near additional degrees, can enable cataract patients to have significantly improved binocular full-range vision, visual function and visual quality. When the degree of reserved near additions in the non-dominant eye preoperatively is>0.20 D and≤0.60 D, it can ensure sufficient binocular UDVA, UIVA and UNVA after surgery, and meanwhile help to obtain superior contrast sensitivity and stereopsis, as well as a satisfactory spectacles-independent rate and low incidence of photic phenomena.

[Contribution of central motion conduction time to the assessment of corticospinal tract lesions and its clinical significance].

Objective: To investigate the association of central motion conduction time (CMCT) with corticospinal tract lesions and its clinical application. Methods: Patients who completed transcranial magnetic stimulation-motor evoked potentials were included from Department of Neurology, Xuanwu Hospital between June 2020 and June 2021. The differences of CMCT values between corticospinal tract sign-positive group and tendon reflex-positive group and the relevant negative groups were compared. The consistency between increased CMCT values and the positive signs of corticospinal tract damage, as well as the significance of CMCT in different neurological diseases were further evaluated. Results: A total of 271 patients were included in the study, aged 12-86 (49±16) years, with 137 males (50.55%) and 134 females (49.45%). The CMCT valuesï¼»M（Q1，Q3）］from Hoffmann's sign-positive group [9.52 (8.54, 10.99) ms vs 9.03 (8.30, 9.53) ms], Babinski's sign-positive group [19.54 (16.97, 24.43) ms vs 16.85(15.63, 18.55) ms] and tendon reflex-positive group [15.38 (9.27, 19.28) ms vs 10.49(8.79, 16.60) ms] were larger than those of relevant negative groups (all P<0.01). In the Babinski sign-positive group, 78.01%(181/232) of the patients had increased CMCT, while in the Hoffmann's sign-positive group, only 26.03%(19/73) of the patients had increased CMCT, indicating that the contribution of CMCT from the lower extremities to the assessment of corticospinal tract lesions was better than that of the upper extremities. With the increase of CMCT values in lower limbs, Babinski sign positive rate increased, the difference was statistically significant(P<0.001). In nervous system diseases, the consistency between CMCT and pathological signs was 75.65% (205/271). Conclusions: The contribution of CMCT from the lower extremities to the assessment of corticospinal tract lesions is superior to that of upper limbs. The higher increase of CMCT values are more reliable for corticospinal tract damage. CMCT has a good concordance with corticospinal tract lesions in some neurological diseases, which can be used to assist clinical diagnosis.

[Anti-inflammatory and repaired effects of non-steroidal anti-inflammatory drugs on human dental pulp cells].

OBJECTIVE: To study the effects of non-steroidal anti-inflammatory drugs (NSAIDs) on anti-inflammation and repair of human dental pulp cells (hDPCs). METHODS: Primary hDPCs from the freshly extracted human third molars were cultured and passaged in vitro, and the following experiments were performed using the 4th-6th generations of hDPCs. HDPCs were cultured in Dulbecco's modified eagle medium (DMEM) containing 1 mg/L lipopolysaccharide (LPS) to obtain LPS irritated hDPCs (LPS-hDPCs), which served as the inflammatory positive group. LPS-hDPCs in the experimental group were cultured in DMEM containing different concentrations (1, 10, and 100 µmol/L) of NSAIDs (aspirin or meloxicam). HDPCs cultured in DMEM were used as the negative control group. The effects of NSAIDs on the proliferation of hDPCs were assessed on the 1st, 3rd, 5th, and 7th day by MTT assay. The effects of NSAIDs on the expression of inflammation related genes interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) of LPS-hDPCs were detected at the 6th hour by real-time PCR. The expression of differentiation related markers dentin matrix protein-1 (DMP-1) and dentin sialophosphoprotein (DSPP) were detected on the 7th day by real-time PCR. The effects of NSAIDs on the mineralization of LPS-hDPCs were assesd on the 14th day by alizarin red staining. Calcium mineralized nodules were semi-quantitatively determined by cetyl pyridine chloride. RESULTS: MTT assay showed that 1-100 µmol/L aspirin or meloxicam significantly promoted the proliferation of hDPC in a concentration dependent manner (P<0.05). Real-time PCR showed that 1-100 µmol/L meloxicam or 100 µmol/L aspirin down-regulated significantly the mRNA expression of TNF-α and IL-6 of LPS-hDPCs (P<0.05), and 100 µmol/L meloxicam down-regulated IL-6 and TNF-α more significantly than 100 µmol/L aspirin of LPS-hDPCs (P<0.05). Real-time PCR showed that 100 µmol/L meloxicam up-regulated the mRNA expression of DMP-1 and DSPP of LPS-hDPCs significantly (P<0.05). Alizarin red staining showed the meloxicam at the concentration of 100 µmol/L significantly promoted the mineralization of LPS-hDPCs (P<0.05). CONCLUSION: In this study, meloxicam promoted the proliferation of hDPCs, inhibited the inflammatory reaction and promoted differentiation and mineralization of hDPCs under LPS irritation. The present results suggest that meloxicam may play a role in anti-inflammation and repair of pulp inflammation.

[Clinical characteristics and influencing factors of patients with novel coronavirus pneumonia combined with liver injury in Shaanxi region].

Objective: To understand the clinical characteristics, change of liver function, influencing factors and prognosis in hospitalized patients with coronavirus disease-19 (COVID-19) combined with liver injury. Methods: The general conditions, biochemical indicators of liver, blood clotting mechanism, routine blood test, UGT1A1 * 28 gene polymorphism and other data of 40 cases with COVID-19 admitted to the isolation ward of Tangdu Hospital were retrospectively analyzed. The clinical characteristics, influencing factors and prognosis of liver injury in patients with liver injury group and those with normal liver function group were compared. The mean of two samples in univariate analysis was compared by t-test and analysis of variance. The counting data was measured by χ(2) tests. The non-normal distribution measurement data were described by the median, and the non-parametric test was used. Statistically significant influencing factors were used as the independent variables in univariate analysis. Multiple logistic regression analysis was used to analyze the main influencing factors of liver injury. Results: Of the 40 cases, 25 were male (62.5%) and 15 were female (37.5%), aged 22 to 83 (53.87 ± 15.84) years. Liver injury was occurred in 22 cases (55%) during the course of the disease. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) level was initially increased (4.4 to 3.5 times of the normal value) along with decrease of albumin in the second week, and the difference was statistically significant (P < 0.001). Ten cases (43.5%) had highest abnormal total blood bilirubin (54.1 µmol/ L). There was no correlation between the increase in transaminase and the increase in total blood bilirubin (R = -0.006, P = 0.972). Three cases had prothrombin activity (PTA) of ≤50%, 10 cases had elevated FDP, and 13 cases had elevated D-dimer, all of whom were severe or critically ill. Liver function injury was more likely to occur in patients who used many types of drugs and large amounts of hormones (P = 0.002, P = 0.031), and there was no correlation with the TA6TA7 mutation in the UGT1A1 * 28 gene locus. Multiple regression analysis showed that the occurrence of liver injury was only related to critical illness. The liver function of all patients had recovered within one week after conventional liver protection treatment. Conclusion: COVID-19 combined with liver function injury may be due to the slight elevation of transaminase, mostly around the second week of the disease course. Severe patients have a higher proportion of liver injury, and critical type is an independent risk factor for liver injury.

Effects of phyto-oestrogen quercetin on productive performance, hormones, reproductive organs and apoptotic genes in laying hens.

Quercetin, a polyphenolic flavonoid with diverse biological activities including anti-inflammatory and antiviral, inhibits lipid peroxidation, prevents oxidative injury and cell death. The purpose of the research was to investigate the effect of quercetin on productive performance, reproductive organs, hormones and apoptotic genes in laying hens between 37 and 45 weeks of age, because of the structure and oestrogenic activities similar to 17ß-oestradiol. The trial was conducted using 240 Hessian laying hens (37 weeks old), housed in wire cages with two hens in each cage. These hens were randomly allotted to four treatments with six replicates, 10 hens in each replicate and fed with diets containing quercetin as 0, 0.2, 0.4 and 0.6 g/kg feed for 8 weeks. The results showed that dietary quercetin significantly increased (p < .05) the laying rate and was higher in group supplemented with 0.4 g/kg, and feed-egg ratio was decreased (p < .05) by quercetin. Dietary quercetin has no effect (p > .05) on average egg weight and average daily feed intake. Compared with control, secretion of hormones, oestradiol (E2 ), progesterone (P4), follicle-stimulating hormone (FSH), luteinizing hormone (LH), insulin-like growth factors-1 (IGF-1) and growth hormone (GH), was found to be significantly higher (p < .05) in quercetin-supplemented groups. Also ovary index, uterus index and oviduct index were not significantly influenced (p > .05) by quercetin, whereas magnum index, isthmus index, magnum length, isthmus length and follicle numbers were significantly increased (p < .05) with quercetin supplementation. Additionally, expression of apoptotic genes was significantly (p < .05) up-regulated or down-regulated by quercetin. These results indicated that quercetin improved productive performance, and its mechanism may be due to the oestrogen-like activities of quercetin.

Gene structure and expression characteristic of a novel odorant receptor gene cluster in the parasitoid wasp Microplitis mediator (Hymenoptera: Braconidae).

Odorant receptors (ORs) expressed in the antennae of parasitoid wasps are responsible for detection of various lipophilic airborne molecules. In the present study, 107 novel OR genes were identified from Microplitis mediator antennal transcriptome data. Phylogenetic analysis of the set of OR genes from M. mediator and Microplitis demolitor revealed that M. mediator OR (MmedOR) genes can be classified into different subfamilies, and the majority of MmedORs in each subfamily shared high sequence identities and clear orthologous relationships to M. demolitor ORs. Within a subfamily, six MmedOR genes, MmedOR98, 124, 125, 126, 131 and 155, shared a similar gene structure and were tightly linked in the genome. To evaluate whether the clustered MmedOR genes share common regulatory features, the transcription profile and expression characteristics of the six closely related OR genes were investigated in M. mediator. Rapid amplification of cDNA ends-PCR experiments revealed that the OR genes within the cluster were transcribed as single mRNAs, and a bicistronic mRNA for two adjacent genes (MmedOR124 and MmedOR98) was also detected in female antennae by reverse transcription PCR. In situ hybridization experiments indicated that each OR gene within the cluster was expressed in a different number of cells. Moreover, there was no co-expression of the two highly related OR genes, MmedOR124 and MmedOR98, which appeared to be individually expressed in a distinct population of neurons. Overall, there were distinct expression profiles of closely related MmedOR genes from the same cluster in M. mediator. These data provide a basic understanding of the olfactory coding in parasitoid wasps.

[Influence of the Arg-Gly-Asp-Ser sequence on the biological effects of bioactive glass on human dental pulp cells].

OBJECTIVE: Positive effects of bioactive glass (BG) on proliferation, mineralization, and differentiation of human dental pulp cells (hDPCs) was already verified in various former studies. The Arg-Gly-Asp-Ser sequence (RGDS) was confirmed of affecting cell adhesion. Before further investigation, the objective of this study is to investigate whether RGDS can affect the effects of BG on the adhesion, proliferation and mineralization of hDPCs. METHODS: hDPCs were harvested from third molars of 18-25-year-old individuals after informed consent. Enzyme digestion technique was used. The 4th to 6th generation of hDPCs were used for all experiments. The cells of the experimental groups were cultured in Dulbecco minimum essential medium (DMEM) containing ionic dissolution products of BG and RGDS of several concentrations (12.5 mg/L, 25.0 mg/L, 50.0 mg/L, 100.0 mg/L, 200.0 mg/L). DMEM containing ionic dissolution products of BG without RGDS was used for cell culture as control group. Cell adhesion was tested 4 h after cell seeding by MTT assay. Cell proliferation was examined at 1, 3, 5, 7, and 9 d after cell seeding by MTT assay. Cell mineralization was investigated on days 14 and 28 by alizarin red staining. After being stained and dried, mineralized nodules were dissolved by cetylpyridinium chloride (CPC) for semi-quantitative test. Results were statistically analyzed by one way ANOVA, SPSS (version 19.0) and P<0.05 was considered to be significant. RESULTS: Cell adhesion in BG group showed no difference from that in DMEM group. Compared with BG group, hDPCs in BG+RGDS groups suggested weaker cell adhesion.When the concentration of RGDS increased, the adhered cell number decreased. hDPCs cultured with BG and RGDS showed lower proliferation activity in the early stage, while no significant difference was observed after 3 d. BG group promoted the mineralization of hDPCs compared with positive control group, negative control group and RGDS group. No significant difference was observed between BG+RGDS group and BG group or between RGDS group and positive control group. CONCLUSION: BG promotes proliferation and mineralization without affecting cell adhesion of hDPCs. Unbounded RGDS inhibits cell adhesion, but has no influence on the positive effects of BG on the proliferation and mineralization of hDPCs.

[Effects of bioactive glass and extracted dentin proteins on human dental pulp cells].

OBJECTIVE: To investigate the proliferation, odontogenic differentiation and mineralization of human dental pulp cells (HDPCs) on bioactive glass(BG) and extracted dentin proteins(EDP). METHODS: Primary HDPCs were isolated from third molars by enzyme digestion and were cultured in Dulbecco's minimum essential medium (DMEM). Then the 4th generation of HDPCs was cultured with DMEM, which contained BG-EDP, BG, and EDP, respectively. Meanwhile HDPCs were cultured in DMEM as control group. Proliferation of HDPCs was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) colorimetric assay. Odontogenic differentiation was determined by alkaline phosphatase (ALP) activity assay and real-time PCR. Mineralization was investigated by Alizarin red staining and cetylpyridinium chloride (CPC) assay. RESULTS: The proliferation of HDPCs was increased significantly in BG-EDP group on 3,7,and 9 d (optical density value: 1.36±0.06, 2.52±0.20, 2.72±0.29) compared with BG (optical density value: 1.20±0.26, 2.33±0.26, 2.50±0.30), EDP(optical density value: 1.13±0.15, 2.10±0.13, 2.38±0.22) and control group (optical density value: 0.84±0.17, 1.84±0.18, 1.95±0.19), P<0.05. After 7 days, ALP activity of BG-EDP group had no statistical difference compared with EDP group and control group; the expression of odontogenic differentiation genes (DSPP, DMP-1) showed no difference among all the groups(P>0.05). After 14 days, ALP activity of BG-EDP group (56.67±1.83) was significantly upregulated compared with EDP group (41.98±9.71) and control group (30.82±6.70), P<0.05, but had no statistical difference compared with BG group (56.29±6.20), P>0.05; DSPP gene expression was upregulated significantly in BG-EDP group (5.79±1.94) compared with the other groups (P<0.05); DMP-1 gene expression of BG-EDP group (3.87±1.87) increased but had no statistical difference compared with the other groups (P>0.05). The alizarin red staining showed more mineral nodules in BG-EDP group, the cetylpyridinium chloride semi-quantification presented higher calcification in BG-EDP group (0.27±0.01) compared with the other groups (P<0.05). CONCLUSION: Compared with either BG or EDP, BG-EDP significantly promotes the proliferation, odontogenic differentiation and mineralization of HDPCs.

Predicting precipitation on nonpoint source pollutant exports in the source area of the Liao River, China.

The source area of the Liao River is an important grain growing area in China which experiences serious problems with agricultural nonpoint source pollution (NPS) which is impacting the regional economy and society. In order to address the water quality issues it is necessary to understand the spatial distribution of NPS in the Liao River source area. This issue has been investigated by coupling a wavelet artificial neural network (WA-ANN) precipitation model with a soil and water assessment tool (SWAT) model to assess the export of nonpoint source pollutants from the Liao River source area. The calibration and validation of these models are outlined. The WA-ANN models and the SWAT model were run to generate the spatial distribution of nonpoint source nutrient (nitrogen and phosphorus) exports in the source area of the Liao River. It was found that the SWAT model identified the sub-catchments which not only receive high rainfall but are also densely populated with high agricultural production from dry fields and paddy fields, which are large users of pesticides and chemical fertilizer, as the primary source areas for nutrient exports. It is also concluded that the coupled WA-ANN models and the SWAT model provide a tool which will inform the identification of NPS issues and will facilitate the identification of management practices to improve the water environments in the source area of the Liao River.

Association between SNPs in genes involved in folate metabolism and preterm birth risk.

We investigated the association between 12 single nucleotide polymorphisms (SNPs) in 11 genes involved in folate metabolic and preterm birth. A subset of SNPs selected from 11 genes/loci involved in the folic acid metabolism pathway were subjected to SNaPshot analysis in a case-control study. Twelve SNPs (CBS-C699T, DHFR-c594+59del19, GST01-C428T, MTHFD-G1958A, MTHFR-C677T, MTHFR-A1298C, MTR-A2756G, MTRR-A66G, NFE2L2-ins1+C11108T, RFC1-G80A, TCN2-C776G, and TYMS-1494del6) in 503 DNA samples were simultaneously tested, and included 315 preterm births and 188 controls. None of the 12 SNP genotype distributions related to the folic acid metabolism pathway showed a significant difference between preterm and term babies. The frequency of the compound mutation genotype of MTHFD-G1958A, MTR-A2756G and RFC1-G80A in preterm babies was 7.3%, which was significantly higher than the 2.7% in term babies. Seven babies carried the compound mutation genotype of MTHFD-G1958A, MTR-A2756G, and CBS-C699T, but this was not observed in term babies. The frequency of the combined wild-type genotype of MTHFD-G1958A, MTR-A2756G, MTRR-A66G, MTHFR-A1298C, NFE2L2-ins1+C11108T, and RFC1- G80A in preterm babies was 3.17%, which was significantly lower than the 7.4% in term babies. The 12 SNPs screened in this study were not independent risk factors of preterm birth. Compound mutation genotypes, including MTHFD-G1958A, MTR-A2756G, and RFC1- G80A and MTHFD-G1958A, MTR-A2756G, and CBS-C699T, may increase the risk of preterm birth. The combined wild-type genotype MTHFD-G1958A, MTR-A2756G, MTRR-A66G, MTHFR-A1298C, NFE2L2-ins1+C11108T, and RFC1-G80A may decrease the risk of preterm birth.

The prediction of 30-day mortality in patients with primary pontine hemorrhage: a scoring system comparison.

BACKGROUND AND PURPOSE: Owing to its low morbidity but high mortality, no accurate scoring system focuses on primary pontine hemorrhage (PPH) has been established. We aim to compare the performances of the Acute Physiology and Chronic Health Evaluation (APACHE) II and the Simplified Acute Physiology Score (SAPS) II with the ICH score in predicting the 30-day mortality in patients with PPH. METHODS: We conducted a retrospective analysis of patients admitted with a diagnosis of PPH to a university-affiliated hospital in southern China from May 2000 to June 2011. Data related to patient demographics and that necessary to calculate APACHE II, SAPS II, and ICH score were recorded. Performances of these scoring systems were presented as calibration and discrimination, which were measured by the Hosmer-Lemeshow goodness-of-fit test and the area under the receiver operating characteristic (ROC) curve, respectively. RESULTS: Among 75 patients with PPH finally included, 31 (41.3%) died within 30 days. SAPS II (χ(2) = 6.57, P = 0.682) had the best calibration, followed by APACHE II (χ(2) = 8.06, P = 0.428) and ICH score (χ(2) = 4.94, P = 0.176). Furthermore, in terms of area under the ROC curve, APACHE II (0.919) was more discriminative than SAPS II (0.890) and ICH score (0.844). CONCLUSIONS: In predicting 30-day mortality in patients with PPH, SAPS II has the best calibration, while APACHE II has the highest discrimination. The ICH score, which is easier and simpler to calculate, should be modified for PPH.

[Intestinal microbiome and its relationship with necrotizing enterocolitis in very low birth weight preterm infants].

Objective: To explore the composition of intestinal microflora prior to onset of necrotizing enterocolitis (NEC) in very low birth weight preterm infants. Methods: This was a multicenter prospective nested case-control study. A total of 46 very low birth weight preterm infants (birth weight <1 500 g and gestional age <35 weeks) within 24 h of life admitted into Neonatal Intensive Care Unit of Children's Hospital of Soochow University and Suzhou Municipal Hospital from April 20 to November 20, 2018 were enrolled. Baseline clinical data and fecal samples of these infants were collected. The subsequent sampling time points were 1st, 4th and 7th day in the first week of life then once per week consecutively. The endpoint of sampling was NEC occurrence, patient discharge or the 8th week post-discharge, whichever came first. Fecal samples were analyzed by 16 S rDNA high-throughput nucleotide sequencing. The control cases were infants without NEC who were matched to the NEC cases with a ratio of 1∶1. The operational taxonomic units (OTU), sequence number and shannon diversity index of the fecal samples were analyzed. Continuous variables were compared with t-test or non-parametric test, and χ2 test or Fisher's exact test was used for categorical variables. Results: There were 23 patients in each group. The gestational age was (29.4±1.8) weeks in NEC group and (29.9±1.6) weeks in control group, including 13 males (57%) and 11 males (48%) in each group, respectively. Species abundance showed that the Firmicutes in both groups decreased temporarily at 7 days of age and then increased with age in control group, but not in NEC group, the Proteobacteria in both groups increased at 7 days of age and then decreased in control group, but kept increasing in NEC group. Regarding the other levels of taxonomy, compared with that of the control group, the NEC group had lower abundance of Proteobacteria, γ-proteobacteria and Enterobacteriaceae at 7 days of age, while higer abundance of Faecalibacterium at 14 days of age, meanwhile, lower Clostridium and Streptococcus at 21 days of age, lower Firmicutes, Clostridia and Clostridium perfringens and higher Proteobacteria and γ-proteobacteria at 28 days of age, these differences were all statistically significant (U=43.00, 43.00, 45.00, 80.00, 74.00, 76.00, 19.00, 8.00, 36.00, 25.00, 25.00,all P<0.05). The shannon index of NEC group was both lower than that of the controls at 21 days of age (2.4 (1.4, 3.0) vs. 3.1 (2.6, 4.0), U=67.00, P=0.027) and 28 days of age (2.4 (1.4, 2.8) vs. 3.9 (3.3, 4.2), U=12.00, P=0.001). Conclusions: The intestinal microflora profile of very low birth weight preterm infants has already changed prior to NEC development. The emergence of differential flora and the reduction of microflora diversity may facilitate early identification and prevention of NEC.

Meishan neonatal piglets tend to have higher intestinal barrier function than crossbred neonatal piglets.

Meishan pigs tend to have higher disease resistance than commercial breeds, although more studies are needed to confirm this difference. This study compared intestinal barrier function between Meishan and crossbred neonatal piglets to provide guidance for both the breeding and nutritional regulation of pigs. Six Meishan piglets and 6 Duroc × (Landrace × Yorkshire) crossbred neonatal piglets (all with normal birth weights) were obtained and allocated into the MEIS and CROSS groups, respectively. Intestinal morphology, goblet cell numbers, antioxidant enzyme activity, and cytokine gene and tight junction protein expression were assessed. The results showed that BW was lower in the MEIS group than in the CROSS group (P < 0.01). The relative lengths of the duodenum (P < 0.05), jejunum (P < 0.01) and ileum (P < 0.01) in the MEIS group were higher than those in the CROSS group. Compared with the CROSS group, the MEIS group exhibited shorter villus lengths in the duodenum and jejunum (P < 0.01), a shallower crypt depth in the ileum (P < 0.001) and denser and longer microvilli in the intestine. The numbers of GCs in the duodenum (P < 0.01) and jejunum (P < 0.001) and the activity levels of glutathione peroxidase (P < 0.05) in the jejunum and of catalase (P < 0.01) and superoxide dismutase (P < 0.01) in the ileum were higher in the MEIS group than in the CROSS group. Compared with the CROSS group, the MEIS group exhibited higher gene expression levels of interleukin (IL) 4 and interferon γ (IFNγ) in the jejunum (P < 0.05); IL2 (P < 0.05), IL4 (P < 0.01) and IFNγ (P < 0.001) in the ileum; and mucin 2 (P < 0.01) and occludin (P < 0.05) in the duodenum. In conclusion, Meishan neonatal piglets showed lower birth weights but higher intestinal barrier function than crossbred piglets.

[Trend analysis and prediction of viral hepatitis incidence in China, 2009-2018].

Objective: To explore the time series characteristics of 5 types of viral hepatitis in China and predict their incidence through effective models. Methods: The monthly incidence data of 5 types of viral hepatitis (A, B, C, D and unspecified) in China from 2009 to 2018 were collected for descriptive and time series analyses, decomposition methods were used to explore the seasonality in the form of seasonal indices and the long-term trend in the form of a linear regression model. Autoregressive integrated moving average (ARIMA) models were established for each type of viral hepatitis. Results: From 2009 to 2018, a total of 14 856 990 cases of viral hepatitis were reported, the seasonal index range of 5 types of viral hepatitis were all lower than 1, the seasonality of hepatitis E was significant, and its incidence was unimodal, but no obvious seasonality characteristics were observed for other four types of viral hepatitis. The incidences of hepatitis A, hepatitis E and unspecified hepatitis remained at lower levels, showing slow declines. Although the cases of hepatitis B accounted for the highest proportion (79.59%, 11 824 262/14 856 990) among 5 types of viral hepatitis, the decline was fastest (-0.01/100 000). The incidence of hepatitis C was on rise, and the rate of increase remained stable (0.005/100 000). The predicted incidences of 5 types of viral hepatitis in China from January 2009 to December 2018 fitted by ARIMA model were consistent with the actual incidences, and the mean absolute error percentage (MAPE) ranged from 3.756 8 to 8.068 3. Conclusions: Time series analysis on surveillance data is useful for better understanding the incidence of the viral hepatitis. The ARIMA model has good application value in the short-term prediction of viral hepatitis incidence in China.

Lnc-RNA BLACAT1 regulates differentiation of bone marrow stromal stem cells by targeting miR-142-5p in osteoarthritis.

OBJECTIVE: Osteogenic differentiation of bone marrow stromal stem cells (BMSCs) is beneficial to the treatment of osteoarthritis (OA). Lnc-RNA BLACAT1 involves in occurrence and development of various diseases. However, the role of Lnc-RNA BLACAT1 in BMSCs differentiation under inflammation remains unclear. MATERIALS AND METHODS: Rat BMSCs were isolated and randomly divided into control group and inflammation group (addition of IL-6). The inflammation group was further divided into BLACAT1 siRNA group and BLACAT1 siRNA+miR-142-5p inhibitor group, followed by analysis of Lnc-RNA BLACAT1 expression by real time PCR, BMSCs proliferation, Caspase 3 activity, ALP activity, expression of Runx2, OC and PPARγ2 by real time PCR, and secretion of TNF-α and IL-1ß by enzyme-linked immunosorbent assay (ELISA). The bioinformatics software and the Luciferase reporter system analyze the targeted relationship between BLACAT1 and miR-142-5p. RESULTS: In inflammation group, Lnc-BLACAT1 expression was increased, along with inhibited BMSCs proliferation, increased Caspase 3 activity, decreased ALP activity, and expression of Runx2 and OC, increased PPARγ2 expression and secretion of TNF-α and IL-1ß. The difference was statistically significant compared with control group (p<0.05). MiR-142-5p is the target miRNA of Lnc-RNA BLACAT1. BLACAT1 siRNA down-regulated BLACAT1 expression, promoted cell proliferation, inhibited Caspase 3 activity, increased ALP activity and Runx2 and OC expression, decreased PPARγ2 expression and TNF-α and IL-1ß secretion. Compared with inflammation group, the difference was statistically significant (p<0.05). Of note, BLACAT1 siRNA+miR-142-5p inhibitor group reversed the effect of siRNA-mediated knockdown of BLACAT1. CONCLUSIONS: Lnc-RNA BLACAT1 expression was increased in inflammatory BMSCs, and knockdown of BLACAT1 promoted proliferation and osteogenic differentiation of BMSCs targeting miR-142-5p.

Effects of rumen-protected betaine supplementation on meat quality and the composition of fatty and amino acids in growing lambs.

Rumen-protected betaine (RPB) can enhance betaine absorption in the small intestine of ruminants, while betaine can alter fat distribution and has the potential to affect the meat quality of livestock. Hence, we hypothesized that RPB might also affect the meat quality of lambs. Sixty male Hu sheep of similar weight (30.47 ± 2.04 kg) were selected and randomly subjected to five different treatments. The sheep were fed a control diet (control treatment, CTL); 1.1 g/day unprotected-betaine supplemented diet (UPB); or doses of 1.1 g/day (low RPB treatment; L-PB), 2.2 g/day (middle RPB treatment; M-PB) or 3.3 g/day (high RPB treatment; H-PB) RPB-supplemented diet for 70 days. Slaughter performance, meat quality, fatty acid and amino acid content in the longissimus dorsi (LD) muscle, shoulder muscle (SM) and gluteus muscle (GM) were measured. Compared with CTL, betaine (including UPB and RPB) supplementation increased the average daily weight gain (ADG) (P < 0.05) and average daily feed intake (P < 0.01) of lambs. Rumen-protected betaine increased ADG (P < 0.05) compared with UPB. With increasing RPB doses, the eye muscle area of the lambs linearly increased (P < 0.05). Compared with CTL, betaine supplementation decreased water loss (P < 0.05) in SM and increased pH24 in the SM (P < 0.05) and GM (P < 0.05). Compared with UPB, RPB decreased water loss in the GM (P < 0.01), decreased shear force (P < 0.05) in the LD and SM and increased the pH of the meat 24 h after slaughter (pH24). With increasing RPB doses, the shear force and b* value in the LD linearly decreased (P < 0.05), and the pH24 of the meat quadratically increased (P < 0.05). Compared with CTL, betaine supplementation increased the polyunsaturated fatty acid in the GM (P < 0.05). Compared with UPB, RPB supplementation decreased the saturated fatty acid (SFA) content in the LD (P < 0.05) and increased the unsaturated fatty acids (UFA), mono-unsaturated fatty acids and UFA/SFA ratio in the LD (P < 0.05). Compared with CTL, the content of histidine in the LD increased with betaine supplementation. Compared with UPB, RPB supplementation increased the content of total free amino acids and flavor amino acids in the LD of lambs (P < 0.05). With increasing RPB, the isoleucine and phenylalanine contents in the LD linearly increased (P < 0.05). Overall, the data collected indicated that the meat quality of lambs (especially in the LD) improved as a result of betaine supplementation, and RPB showed better effects than those of UPB.

Biodegradation of nicotine by a newly isolated Agrobacterium sp. strain S33.

AIMS: To isolate and characterize bacteria capable of degrading nicotine from the rhizospheric soil of a tobacco plant and to use them to degrade the nicotine in tobacco solid waste. METHODS AND RESULTS: A bacterium, strain S33, was newly isolated from the rhizospheric soil of a tobacco plant, and identified as Agrobacterium sp. based on morphology, physiological tests, Biolog MicroLog3 4.20 system and 16S rRNA gene sequence. Using nicotine as the sole source of carbon and nitrogen in the medium, it grew optimally with 1.0 g l(-1) of nicotine at 30 degrees C and pH 7.0, and nicotine was completely degraded within 6 h. The resting cells prepared from the glucose-ammonium medium or LB medium could not degrade nicotine within 10 h, while those prepared from the nicotine medium could completely degrade 3 g l(-1) of nicotine in 1.5 h at a maximal rate of 1.23 g nicotine h(-1) g(-1) dry cell. Using the medium containing nicotine, glucose and ammonium simultaneously to cultivate strain S33, the resting cells could degrade 98.87% of nicotine in tobacco solid waste with the concentration as 30 mg nicotine g(-1) dry weight tobacco solid waste within 7 h at a maximal rate of 0.46 g nicotine h(-1) g(-1) dry cell. CONCLUSIONS: This is the first report that Agrobacterium sp. has the ability to degrade nicotine. Agrobacterium sp. S33 could use nicotine as the sole source of carbon and nitrogen. The use of resting cells of the strain S33 prepared from the nicotine-glucose-ammonium medium was an effective method to degrade nicotine and detoxify tobacco solid waste. SIGNIFICANCE AND IMPACT OF THE STUDY: Nicotine in tobacco wastes is both toxic and harmful to human health and the environment. This study showed that Agrobacterium sp. S33 may be suitable for the disposal of tobacco wastes and reducing the nicotine content in tobacco leaves.

MicroRNA-92a regulates the development of cutaneous malignant melanoma by mediating FOXP1.

OBJECTIVE: MicroRNAs are noncoding RNAs which are involved in the occurrence and progression of tumors. This study aims to explore the role of microRNA-92a in cutaneous malignant melanoma (CMM) and its underlying mechanism. PATIENTS AND METHODS: The expression level of microRNA-92a in 75 pairs of CMM tissues and paracancerous tissues was determined using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between microRNA-92a expression with clinical data of CMM patients was analyzed. Besides, microRNA-92a expression in CMM cells and primary epidermal melanocytes (PEM) was determined by qRT-PCR as well. After transfection of si-microRNA-92a in CMM cells, biological performances of CMM were determined using cell counting kit-8 (CCK-8), colony formation and transwell assay, respectively. FOXP1 expression in CMM cells and tissues was determined using Western blot. Kaplan-Meier survival curves were drawn to explore the correlation between the FOXP1 expression and prognosis of CMM patients. RESULTS: MicroRNA-92a was highly expressed in CMM tissues compared with that of paracancerous tissues. Compared with CMM patients with lower expression of microRNA-92a, those with higher expression of microRNA-92a presented higher tumor stage, higher incidences of lymph node metastasis and distant metastasis, as well as lower overall survival. The knockdown of microRNA-92a remarkably decreased proliferative, invasive and metastatic capacities of CMM cells. Western blot results elucidated that microRNA-92a knockdown in CMM cells upregulates FOXP1 expression. Additionally, rescue experiments showed that mi-croRNA-92a regulates biological performances of CMM cells by regulating FOXP1. CONCLUSIONS: MicroRNA-92a is highly expressed in CMM, which is remarkably correlated to tumor stage and poor prognosis of CMM patients. We found that microRNA-92a pro-motes malignant progression of CMM by regulating FOXP1.