Antibacterial and Anti-Inflammatory Polysaccharide from Fructus Ligustri Lucidi Incorporated in PVA/Pectin Hydrogels Accelerate Wound Healing.

In cutaneous wound healing, an overproduction of inflammatory chemokines and bacterial infections impedes the process. Hydrogels can maintain a physiologically moist microenvironment, absorb chemokines, prevent bacterial infection, inhibit bacterial reproduction, and facilitate wound healing at a wound site. The development of hydrogels provides a novel treatment strategy for the entire wound repair process. Here, a series of Fructus Ligustri Lucidi polysaccharide extracts loaded with polyvinyl alcohol (PVA) and pectin hydrogels were successfully fabricated through the freeze-thaw method. A hydrogel containing a 1% mixing weight ratio of FLL-E (named PVA-P-FLL-E1) demonstrated excellent physicochemical properties such as swellability, water retention, degradability, porosity, 00drug release, transparency, and adhesive strength. Notably, this hydrogel exhibited minimal cytotoxicity. Moreover, the crosslinked hydrogel, PVA-P-FLL-E1, displayed multifunctional attributes, including significant antibacterial properties, earlier re-epithelialization, production of few inflammatory cells, the formation of collagen fibers, deposition of collagen I, and faster remodeling of the ECM. Consequently, the PVA-P-FLL-E1 hydrogel stands out as a promising wound dressing due to its superior formulation and enhanced healing effects in wound care.

A long transcript mutant of the rubisco activase gene RCA upregulated by the transcription factor Ghd2 enhances drought tolerance in rice.

The transcription factor Ghd2 increases rice yield potential under normal conditions and accelerates leaf senescence under drought stress. However, its mechanism on the regulation of leaf senescence under drought stress remains unclear. In the present study, to unveil the mechanism, one target of Ghd2, the Rubisco activase gene RCA, was identified through the combined analysis of Ghd2-CRISPR transcriptome data and Ghd2-overexpression microarray data. Ghd2 binds to the 'CACA' motif in the RCA promoter by its CCT domain and upregulates RCA expression. RCA has alternative transcripts, RCAS and RCAL, which are predominantly expressed under normal conditions and drought stress, respectively. Similar to Ghd2-overexpressing plants, RCAL-overexpressing plants were more sensitive to drought stress than the wild-type. However, the plants overexpressing RCAS showed a weak drought-sensitive phenotype. Moreover, RCAL knockdown and knockout plants did not show yield loss under normal conditions, but exhibited enhanced drought tolerance and delayed leaf senescence. The chlorophyll content, the free amino acid content and the expression of senescence-related genes in the RCAL mutant were lower than those in the wild-type plants under drought stress. In summary, Ghd2 induces leaf senescence by upregulating RCAL expression under drought stress, and the RCAL mutant has important values in breeding drought-tolerant varieties.

[Protective effect of oleanolic acid liposomes on cisplatin-induced oligoasthenospermia in mice].

OBJECTIVE: To study the protective effect of oleanolic acid liposomes (OA-Lips) on cisplatin-induced oligoasthenospermia (COAS) in mice. METHODS: Sixty ICR mice were randomly divided into a normal control, a COAS model control, a positive control and a low-, a medium- and a high-dose OA-Lips group. The animals in the low-, medium- and high-dose OA-Lips and positive control groups were given intragastrically OA-Lips solution at 25, 50, and 100 mg/kg/d and vitamin E at 50 mg/kg/d, respectively. On the 28th day, the mice in the COAS model control, positive control and OA-Lips groups were injected intraperitoneally with cisplatin solution at 10 mg/kg, while those in the normal control group with the same dose of normal saline. Three days after administration, all the mice were sacrificed and their testis tissues collected for detection of the semen parameters and observation of the testicular morphology. RESULTS: Both the percentage of motile sperm and sperm concentration were significantly increased in the high-dose OA-Lips group (P < 0.05). HE staining showed that OA-Lips remarkably improved the damaged testis tissue (P < 0.05) and protected the seminiferous tubules and interstitial cells. The percentage of progressively motile sperm (PMS) and the curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN), straightness (STR), wobble (WOB), amplitude of lateral head displacement (ALH) and beat-cross frequency (BCF) of sperm were gradually increased in a dose-dependent manner in the OA-Lips groups. The serum T level was significantly higher (P < 0.05) in the OA-Lips-treated mice than in the COAS model controls while the percentage of morphologically abnormal sperm (MAS) markedly lower in the high-dose OA-Lips group than in the model control, positive control and low-dose OA-Lips groups (P < 0.05). CONCLUSION: OA-Lips can relieve oligoaspermia and improve the productive ability of mice.

Influence of Different Inactivation Methods on Severe Acute Respiratory Syndrome Coronavirus 2 RNA Copy Number.

The outbreak of coronavirus disease 2019 (COVID-19) has spread across the world and was characterized as a pandemic. To protect medical laboratory personnel from infection, most laboratories inactivate the virus causing COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), in clinical samples before testing. However, the effect of inactivation on the detection results remains unknown. Here, we used a digital PCR assay to determine the absolute SARS-CoV-2 RNA copy number in 63 nasopharyngeal swab samples and assess the effect of inactivation methods on viral RNA copy number. Viral inactivation was performed by three different methods: (i) incubation with the TRIzol LS reagent for 10 min at room temperature, (ii) heating in a water bath at 56°C for 30 min, and (iii) high-temperature treatment, including autoclaving at 121°C for 20 min, boiling at 100°C for 20 min, and heating at 80°C for 20 min. Compared to the amount of RNA in the original sample, TRIzol treatment destroyed 47.54% of the nucleocapsid protein (N) gene and 39.85% of open reading frame (ORF) 1ab. For samples treated at 56°C for 30 min, the copy number of the N gene and ORF 1ab was reduced by 48.55% and 56.40%, respectively. The viral RNA copy number dropped by 50 to 66% after heating at 80°C for 20 min. Nearly no viral RNA was detected after autoclaving at 121°C or boiling at 100°C for 20 min. These results indicate that inactivation reduced the quantity of detectable viral RNA and may cause false-negative results, especially in weakly positive cases. Thus, use of the TRIzol reagent rather than heat inactivation is recommended for sample inactivation, as the TRIzol reagent had the least effect on the RNA copy number among the tested methods.

[Myricetin reduces the reproductive toxicity of cyclophosphamide in male mice].

OBJECTIVE: To explore the effect of myrica flavone on male reproductive toxicity induced by cyclophosphamide and its mechanism. METHODS: Thirty 6-week-old male ICR mice were randomly divided into 5 groups: blank control group, cyclophosphamide reproductive injury model group, myricetin low-medium high-dose intervention group. Except the blank control group, the other groups were intraperitoneally injected with cyclophosphamide 50 mg/kg daily for 7 consecutive days. The myricetin group received intragastric administration of 100, 200, and 400 mg/kg myricetin daily for 30 consecutive days since the second day of modeling. The blank control group and the model control group were given an equal volume of a 0. 25% sodium carboxymethyl cellulose solution. The body weight was measured every 3 days, and the day after the last administration, the mice were sacrificed by cervical dislocation, and the epididymis and testes were quickly taken. Testicular weighing, testicular index calculation, epididymis to obtain sperm, sperm analyzer to analyze sperm density and vitality. The expression of Bax and Bcl-2 in testicular tissues was detected by immunoblotting, and the mitochondrial membrane potential of sperm was detected by flow cytometry. RESULTS: After 9 days of modeling, the weight of mice in the model group was lower than that of the blank control group, which was statistically different(P<0. 05). There was no difference between the myricetin treatment group and the model group. The testis index of the model group was(3. 93±0. 91)mg/g, which was significantly lower than that of the blank control group(6. 93±0. 98)mg/g, and the difference was statistically significant(P<0. 05). After treatment with bayberry flavonoids, the testis index increased, in the 100 and 200 groups and 400 mg/kg testis index were(3. 94±1. 21) mg/g, (4. 33±0. 88) mg/g, and(4. 80±0. 43) mg/g, respectively. Compared with model control group, The difference was statistically significant(P<0. 05 and P<0. 01). Compared with the control group, the sperm density, sperm rate of forward movement, sperm rate of non-forward movement, and decreased sperm rate of non-moving sperm increased in the model group. After treatment with bayberry flavonoids, compared with the model group, the sperm density, sperm rate of forward motion, and sperm rate of non-forward motion increased, and the immobility sperm rate decreased. The 200 and 400 mg/kg groups had statistical significance(P<0. 05 or P<0. 01); the normal rate of sperm mitochondrial membrane potential in the model group was(54. 70±5. 45)%, and the normal mitochondrial membrane potential rate after treatment with myricetin of 100, 200 and 400 mg/kg(59. 10±9. 97)%, (62. 10±6. 07)% and(77. 10±8. 87)%, of which the 400 mg/kg group was statistically significant(P<0. 05); the ratio of Bax/Bcl-2 in the model group was 5. 92±1. 45, and the ratio of Bax/Bcl-2 decreased after treatment with myricetin of 100, 200 and 400 mg/kg, which were 2. 52±0. 51, 1. 71±0. 52 and 1. 07±0. 29. There were statistical differences(P<0. 05 or P<0. 01). CONCLUSION: Myrica flavone can protect sperm mitochondrial membrane potential, inhibit testicular cell apoptosis, and protect the male mice from reproductive toxicity induced by cyclophosphamide.

Imaging of Moyamoya Disease and Moyamoya Syndrome: Current Status.

Moyamoya disease (MMD) and Moyamoya syndrome (MMS) are referring to a progressive steno-occlusive vasculopathy at terminal portions of the bilateral internal carotid arteries and their proximal branches with prominent collateral artery formation. They can be found throughout the world and cause irreversible damage to the cerebral hemodynamics due to the progressive nature. Prompt diagnosis and accurate assessment could significantly improve the prognosis of MMD and MMS. Some imaging modalities could be used for diagnosis and nonquantitative evaluation of MMD and MMS, such as conventional computed tomography (CT) and magnetic resonance imaging (MRI), digital subtraction angiography, CT angiography (CTA), and magnetic resonance angiography. Some could quantitatively evaluate the cerebral hemodynamics of MMD and MMS, such as single-photon emission CT, positron emission tomography, xenon-enhanced CT, perfusion CT, dynamic susceptibility contrast MRI, arterial spin labeling MRI, and the hemodynamic parameters measured by those imaging methods could guide treatment of MMD and MMS. All the imaging modalities have their merits and demerits, and they can play a part in certain situation. We need establish standardized protocols for preoperative and postoperative evaluation with different imaging techniques in the further science for MMD and MMS.

Cross reactivity of Neisseria meningitidis crgA diagnostic PCR primers with nontypeable haemophilus influenzae.

BACKGROUND: CrgA based PCR are usually used for diagnosis of Neisseria meningitidis while the crgA gene was observed in the genomes of Haemophilus influenzae. In this study, we aimed to evaluate whether the crgA primers routinely used in the diagnosis of N. meningitidis could cross react with H. influenzae isolates. METHODS: A diagnostic test study analysis of sixty-two H. influenzae isolates from oropharyngeal swabs of healthy individuals aged 9 to 11 years between 2011 and 2012, using commonly used crgA primers for diagnostic analysis of N. meningitidis. RESULTS: It revealed that 19.3% nontypable H. influenzae isolates were positive for the crgA gene. All the biotype IV H. influenzae isolates were crgA PCR positive. CONCLUSIONS: Our study has shown a significant finding of crgA gene especially in bitotype IV nontypable H. influenzae by N. meningitidis crgA diagnostic PCR primers. It is necessary to further evaluate the prevalence of the crgA gene in more non-typable H. influenzae strains, particularly invasive strains.

The advantages of the magnetic resonance image compilation (MAGiC) method for the prognosis of neonatal hypoglycemic encephalopathy.

Objectives: To explore the prognostic value of magnetic resonance image compilation (MAGiC) in the quantitative assessment of neonatal hypoglycemic encephalopathy (HE). Methods: A total of 75 neonatal HE patients who underwent synthetic MRI were included in this retrospective study. Perinatal clinical data were collected. T1, T2 and proton density (PD) values were measured in the white matter of the frontal lobe, parietal lobe, temporal lobe and occipital lobe, centrum semiovale, periventricular white matter, thalamus, lenticular nucleus, caudate nucleus, corpus callosum and cerebellum, which were generated by MAGiC. The patients were divided into two groups (group A: normal and mild developmental disability; group B: severe developmental disability) according to the score of Bayley Scales of Infant Development (Bayley III) at 9-12 months of age. Student's t test, Wilcoxon test, and Fisher's test were performed to compare data across the two groups. Multivariate logistic regression was used to identify the predictors of poor prognosis, and receiver operating characteristic (ROC) curves were created to evaluate the diagnostic accuracy. Results: T1 and T2 values of the parietal lobe, occipital lobe, center semiovale, periventricular white matter, thalamus, and corpus callosum were higher in group B than in group A (p < 0.05). PD values of the occipital lobe, center semiovale, thalamus, and corpus callosum were higher in group B than in group A (p < 0.05). Multivariate logistic regression analysis showed that the duration of hypoglycemia, neonatal behavioral neurological assessment (NBNA) scores, T1 and T2 values of the occipital lobe, and T1 values of the corpus callosum and thalamus were independent predictors of severe HE (OR > 1, p < 0.05). The T2 values of the occipital lobe showed the best diagnostic performance, with an AUC value of 0.844, sensitivity of 83.02%, and specificity of 88.16%. Furthermore, the combination of MAGiC quantitative values and perinatal clinical features can improve the AUC (AUC = 0.923) compared with the use of MAGiC or perinatal clinical features alone. Conclusion: The quantitative values of MAGiC can predict the prognosis of HE early, and the prediction efficiency is further optimized after being combined with clinical features.

Whole-volume ADC histogram of the brain as an image biomarker in evaluating disease severity of neonatal hypoxic-ischemic encephalopathy.

Purpose: To examine the diagnostic significance of the apparent diffusion coefficient (ADC) histogram in quantifying neonatal hypoxic ischemic encephalopathy (HIE). Methods: An analysis was conducted on the MRI data of 90 HIE patients, 49 in the moderate-to-severe group, and the other in the mild group. The 3D Slicer software was adopted to delineate the whole brain region as the region of interest, and 22 ADC histogram parameters were obtained. The interobserver consistency of the two radiologists was assessed by the interclass correlation coefficient (ICC). The difference in parameters (ICC > 0.80) between the two groups was compared by performing the independent sample t-test or the Mann-Whitney U test. In addition, an investigation was conducted on the correlation between parameters and the neonatal behavioral neurological assessment (NBNA) score. The ROC curve was adopted to assess the efficacy of the respective significant parameters. Furthermore, the binary logistic regression was employed to screen out the independent risk factors for determining the severity of HIE. Results: The ADCmean, ADCmin, ADCmax,10th-70th, 90th percentile of ADC values of the moderate-to-severe group were smaller than those of the mild group, while the group's variance, skewness, kurtosis, heterogeneity, and mode-value were higher than those of the mild group (P < 0.05). All the mentioned parameters, the ADCmean, ADCmin, and 10th-70th and 90th percentile of ADC displayed positive correlations with the NBNA score, mode-value and ADCmax displayed no correlations with the NBNA score, the rest showed negative correlations with the NBNA score (P < 0.05). The area under the curve (AUC) of variance was the largest (AUC = 0.977; cut-off 972.5, sensitivity 95.1%; specificity 87.8%). According to the logistic regression analysis, skewness, kurtosis, variance, and heterogeneity were independent risk factors for determining the severity of HIE (OR > 1, P < 0.05). Conclusions: The ADC histogram contributes to the HIE diagnosis and is capable of indicating the diffusion information of the brain objectively and quantitatively. It refers to a vital method for assessing the severity of HIE.

MicroRNA-221-5p Promotes Ricin Toxin-induced Inflammation via PI3K/Akt signaling pathway by targeting COL4a5.

Ricin toxin (RT) is one of the most lethal type II ribosome-inactivating proteins (RIP), and is classified as a potential bioterror agent due to its severe cytotoxicity and high availability. The toxicity of RT is dependent on both dose and route of exposure. Increasing evidence demonstrates that sub-lethal RT induces acute inflammation and increases the release of pro-inflammatory cytokines. However, current studies on mechanism of RT-induced inflammation are limited. In this study, to evaluate the relationship between miRNAs and RT-induced inflammation, RNA sequencing (RNA-Seq) was used to analyze the expression of miRNAs and mRNAs in RT-treated RAW264.7 macrophage cells. A total of 14 significantly differently expressed (DE) miRNAs and 323 miRNA-mRNA interaction pairs were predicted by bioinformatics analysis. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that majority of those interaction pairs were involved in PI3K/Akt pathway. In addition, overexpression of miR-221-5p promoted the inflammatory response by inhibiting the mRNA expression of COL4a5. This work contributes to our understanding of RT-induced inflammation and demonstrates the potential role of miRNAs in innate immunity, which may be regarded as potential targets in developing therapies for RT poisoning.

Determination of pyrethroid pesticide residues in vegetables by solvent sublation followed by high-performance liquid chromatography.

A novel method is developed for separating and enriching pyrethroid pesticides from vegetables by solvent sublation, and determination of the pyrethroids is performed by high-performance liquid chromatography (HPLC). The effects of organic solvent, pH of the solution, nitrogen flow rate, and sublation time on the sublation efficiency of pyrethroids are investigated in detail, and the optimal conditions of the solvent sublation are selected. The floated product of vegetables in the optimal conditions is determined by HPLC. The limit of detection values range from 1.4 microg/kg (for bifenthrin) to 4.2 microg/kg (for fenpropathin). The recoveries of spiked vegetable samples are from 85.7% to 110.4%, and relative standard deviation values are from 1.70% to 6.19%. The results are satisfactory.

Application of solvent sublation for the determination of organophosphorous pesticides in vegetables by gas chromatography with a flame photometric detector.

An effective method for separating and enriching organophosphorous pesticides (OPPs) from vegetables by solvent sublation and the OPPs determination by gas chromatography with flame photometric detector (GC-FPD) has been developed. The effects of organic solvent, nitrogen flow rate, pH of the solution, sublation time etc. on the sublation efficiency of OPPs were investigated in detail, and the optimal conditions of solvent sublation were selected. The floated product of vegetables in the optimal conditions was determined by GC-FPD. The limits of detection (LODs) ranged from 1.2 microg kg-1 (for dimethoate) to 3.5 microg kg-1 (for chlorpyrifos). The recoveries of spiked vegetable samples were from 81.3 to 98.9%, and RSD values were from 0.46 to 4.83%. The results show that this method is simple, sensitive and rapid.

[Recognizing the vaccination strategy of pertussis according to the family aggregation feature of transmission].

OBJECTIVE: To understand the age distribution of pertussis patients admitted in the children hospital and to analyze the source of infection as well as its transmission patterns. METHODS: Patients visiting to the Children Hospital and epidemiologically related cases during Feb. 2012 to Aug. 2013 were tested to confirm the diagnosis. Excel 2007 software was used to analyze the age distribution and clinical symptoms of clinic cases, the source of infection or subsequent cases. RESULTS: 165 out of 254 clinically suspicious pertussis cases and 38 out of the 54 epidemiologically related cases were confirmed of having pertussis infection. There were 138 (83.6%) cases under 1 year of age in the confirmed clinical cases and 36 (94.7%) cases older than 20 years of age among the confirmed epidemiologically related pertussis cases. All the confirmed epidemiologically related cases were misdiagnosed or missed for diagnosis. As the source of pertussis infection in confirmed clinical cases, parents played an imported role among 25 of the 32 cases. Transmission from infants and/or little children to adults were also observed in this study. CONCLUSION: Infants accounted for the most among the pertussis patients that visiting the clinics. Adults, being misdiagnosed or missed diagnosed, were the main sources of infection to infants. Epidemics of pertussis occurred under family aggregation. Further study was in need to develop the proper strategy for pertussis booster vaccination.

Scab-inspired cytophilic membrane of anisotropic nanofibers for rapid wound healing.

This work investigates the influence of cytophilic and anisotropic nanomaterials on accelerated cell attachment and directional migration toward rapid wound healing. Inspired by the anisotropic protein nanofibers in scab, a polyurethane (PU) nanofibrous membrane with an aligned structure was fabricated. The membrane showed good affinity for wound-healing-related cells and could guide cell migration in the direction of PU nanofibers. Also, the morphology and distribution of F-actin and paxillin of attached cells were influenced by the underlying nanofibers. The randomly distributed PU nanofibers and planar PU membrane did not show a distinct impact on cell migration. This scab-inspired cytophilic membrane is promising in applications as functional interfacial biomaterials for rapid wound healing, bone repair, and construction of neural networks.

A triggered DNA hydrogel cover to envelop and release single cells.

We develop an enzyme-triggered permeable DNA hydrogel cover to envelop and release single cells in microwells. The porous structure of the DNA hydrogel allows nutrients and waste to pass through, leading to a cell viability as high as 98%. The design provides a general method to culture, monitor, and manipulate single cells, and has potential applications in cell patterning and studying cell communication.

[Comparative studies on different methods for laboratory diagnosis of pertussis].

OBJECTIVE: To confirm the clinically suspected pertussis cases(<1 years old) through laboratory methods. METHODS: From December, 2011 to December, 2012, patients with clinically suspected pertussis from Xi'an Children's Hospital were sampled, with their nasopharyngeal swabs collected, blood samples cultured and pertussis toxin IgG detected by PCR. RESULTS: were analyzed, using SPSS 16.0 software. RESULTS: 100 out of the 148 cases were laboratorial confirmed. 3, 88 and 34 cases were positive, through culture, PCR or pertussis toxin IgG respectively. 22 cases were both PCR and pertussis toxin IgG positive. There were significant differences between the results of IS481 PCR, days from the onset of symptoms (P < 0.01) and results of PT-IgG with the days from onset of symptoms (P < 0.01). CONCLUSION: Since the sensitivity of culture on pertussis was low, diagnosis on the disease should be linked to the results from PCR, PT-IgG and the days from onset of symptoms.