Research on genetic model of keratoconus / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To explore the effect of genetic factors on the pathogenesis of keratoconus and its genetic model.METHODS: Genetic epidemiological methods were used to investigate the prevalence of keratoconus in 280 first-degree relatives of 100 patients with keratoconus who attended Henan Eye Hospital between July 2020 and April 2023. The heritability was estimated by Falconer regression method. The general genetic model was calculated using Penrose method, and the genetic model was confirmed by Falconer formula, Edwards approximation formula and the projection formula of San-Duo Jiang's threshold model theory.RESULTS: The results showed that there were 16(5.714%)first-degree relatives of keratoconus probands suffering from keratoconus, and the heritability of keratoconus was(86.100±7.400)%. The S/q score calculated by the Penrose method was 35.348, which was near to 1/(q)1/2, suggesting that the genetic model of keratoconus might be polygenic inheritance. The expected prevalence in first-degree relatives of keratoconus patients by Falconer formula, Edwards approximation formula and the projection formula of San-Duo Jiang's threshold model theory were 5.900%, 7.714% and 5.700%, respectively, which showed no significant differences from the actual prevalence(5.714%), suggesting that keratoconus was a polygenetic disease.CONCLUSION:Genetic factors might play an important role in the pathogenesis of keratoconus, and keratoconus is a polygenetic disease.

Clinical features of keratoconus and influencing factors of disease severity / 中华实验眼科杂志

Objective:To investigate the clinical characteristics of patients with keratoconus, and to explore the factors influencing keratoconus severity.Methods:A cross-sectional study was performed.A total of 908 patients (1 476 eyes) with primary keratoconus were enrolled in Henan Eye Hospital from January 2019 to December 2021.The medical history data of patients were collected by face-to-face questionnaire survey.Refractive parameters were measured by subjective optometry.Intraocular pressure (IOP) was measured by a non-contact tonometer, and corrected IOP was calculated by Dresden formula.Corneal topography parameters was obtained using Pentacam HR.The subgroup analysis of clinical characteristics of all patients was performed by age (<21 years, 21~<31 years, ≥31 years) and gender.Disease severity was graded based on steep keratometry (Ks), namely mild (Ks<48 D), moderate (48 D≤Ks<55 D) and severe (Ks≥55 D). The influencing factors of disease severity in keratoconus were analyzed by ordered Logistic regression.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2019[5]). All subjects or guardians were informed of the purpose and significance of the study and written informed consent was obtained.Results:Of the 908 patients, 622 were with bilateral keratoconus and 286 were with unilateral keratoconus.The median age of onset was 20(17, 26) years, and the median age of diagnosis was 21(18, 27) years.The ratio of males to females was 3.05∶1.There were 9.80%(89/908) of the patients having a history of allergy, 25.55%(232/908) having a history of other systemic diseases, and 1.98%(18/908) having a family history of keratoconus.Of the 1 476 affected eyes, 27.57%(407/1 476) were diagnosed as severe keratoconus, and 61.94%(568/917) had a history of eye rubbing.The medians of sphericity, cylindricity, IOP, corrected IOP, Ks, thinnest corneal thickness (TCT), anterior corneal surface elevation (AE) and posterior corneal surface elevation (PE) were -4.00(-7.00, -1.75)D, -3.50(-6.00, -1.50)D, 12.00(10.30, 13.80)mmHg, 15.40(13.60, 17.00)mmHg, 49.85(46.40, 54.90)D, 460.00(425.00, 490.00)μm, 21.00(13.00, 34.75)μm, 51.00(33.00, 75.00)μm, respectively.The spherical refraction, IOP and corrected IOP were lower and the cylindrical refraction was higher in patients at age <21 years than in patients at age 21~<31 years, and the TCT of patients at age <21 years was higher than that at age ≥31 years, and the differences were statistically significant (all at P<0.05). Compared with female patients, male patients had younger onset age, lower spherical refraction, IOP and corrected IOP, as well as higher cylindrical refraction, AE and PE, showing statistically significant differences (all at P<0.05). The spherical refraction and IOP of male patients were lower than those of female patients at age <21 years, and the cylindrical refraction was higher in males than in females among the patients at age 21~<31 years, and the differences were statistically significant (both at P<0.05). Among the patients with onset age <21 years and diagnosis age <21 years, the ratio of males to females in patients with severe keratoconus was higher than those with mild and moderate disease, and the difference was statistically significant (both at P<0.05). Older age of onset was a protective factor for disease severity in keratoconus (odds ratio=0.981, 95% confidence interval: 0.963~0.999). Conclusions:The younger the onset age of keratoconus patients, the more severe the disease.Among the patients with severe keratoconus, there were more male patients, and males have a younger onset age and severer conditions.It is suggested that early screening of keratoconus in children and adolescents should be strengthened in clinical work, and more active prevention and treatment measures should be taken for younger patients, especially males.

Construction and evaluation of the functional polygenic risk score for gastric cancer in a prospective cohort of the European population / 中华医学杂志(英文版)

BACKGROUND@#A polygenic risk score (PRS) derived from 112 single-nucleotide polymorphisms (SNPs) for gastric cancer has been reported in Chinese populations (PRS-112). However, its performance in other populations is unknown. A functional PRS (fPRS) using functional SNPs (fSNPs) may improve the generalizability of the PRS across populations with distinct ethnicities.@*METHODS@#We performed functional annotations on SNPs in strong linkage disequilibrium (LD) with the 112 previously reported SNPs to identify fSNPs that affect protein-coding or transcriptional regulation. Subsequently, we constructed an fPRS based on the fSNPs by using the LDpred2-infinitesimal model and then analyzed the performance of the PRS-112 and fPRS in the risk prediction of gastric cancer in 457,521 European participants of the UK Biobank cohort. Finally, the performance of the fPRS in combination with lifestyle factors were evaluated in predicting the risk of gastric cancer.@*RESULTS@#During 4,582,045 person-years of follow-up with a total of 623 incident gastric cancer cases, we found no significant association between the PRS-112 and gastric cancer risk in the European population (hazard ratio [HR] = 1.00 [95% confidence interval (CI) 0.93-1.09], P = 0.846). We identified 125 fSNPs, including seven deleterious protein-coding SNPs and 118 regulatory non-coding SNPs, and used them to construct the fPRS-125. Our result showed that the fPRS-125 was significantly associated with gastric cancer risk (HR = 1.11 [95% CI, 1.03-1.20], P = 0.009). Compared to participants with a low fPRS-125 (bottom quintile), those with a high fPRS-125 (top quintile) had a higher risk of incident gastric cancer (HR = 1.43 [95% CI, 1.12-1.84], P = 0.005). Moreover, we observed that participants with both an unfavorable lifestyle and a high genetic risk had the highest risk of incident gastric cancer (HR = 4.99 [95% CI, 1.55-16.10], P = 0.007) compared to those with both a favorable lifestyle and a low genetic risk.@*CONCLUSION@#These results indicate that the fPRS-125 derived from fSNPs may act as an indicator to measure the genetic risk of gastric cancer in the European population.

Application of Tiaoshen Jianpi acupuncture and moxibustion in hospice care for terminal cancer patients / 中国针灸

To analyze the application feasibility of Tiaoshen Jianpi acupuncture and moxibustion in hospice care for terminal cancer patients. Tiaoshen Jianpi acupuncture and moxibustion adjusts the spirit to regulate emotions and fortifies the spleen to supplement and boost foundation of acquired (postnatal) constitution. And it could relieve adverse reactions after radiotherapy and chemotherapy, alleviate pain and regulate emotions in hospice care for terminal cancer patients, so as to promote the progress of hospice care for terminal cancer patients.

Exploration of mechanism of Huoxue Jiedu Chinese Herbal Compatibility in anti-atherosclerosis: Based on network pharmacology and molecular docking / 中国药理学通报

Aim To explore the mechanism of Huoxue Jiedu Chinese Herbal Compatibility (angelica sinensis - lonicerae japonicae flos ) in anti-atherosclerosis by means of network pharmacology and molecular rloe- king.Methods The effective eomponents and related targets of angelica sinensis and lonicerae japonicae flos were sereened by the TCMSP database, and the dis¬ease related targets of atheroselerosis were obtained by using the Gene Cards database, DrugBank database and OMIM database.The common targets were ob¬tained by Venny2.1 , an online mapping tool.The pro- tein-protein interaction ( PPI) network was established by String 11.0, and the common target was analyzed by gene ontology(GO) enrichment analysis and kvoto en- cyelopedia of genes and genomes ( KEGG) enrichment analysis using metascape online platform.Cyto- seape3.8.2 was used to construct the " active ingredi¬ent-active target-pathway network map.Finally AutoDock and PyMOL softwares were applied for mo¬lecular docking.Results A total of 31 active compo¬nents of angelica sinensis-lonicerae japonicae flos com¬ patibility and 509 component targets were screened.Hie number of intersection targets with atherosclerosis was 210.GO function enrichment analysis revealed 2 961 biological process items and 242 molecular func¬tion items.KEGG pathway enrichment screened 250 signaling pathways, PI3K-Akt signaling pathway, MAPK signaling pathway, and fluid shear stress and atherosclerosis were the key pathways.Molecular doc¬king results showed that beta-sitosterol, ferulic acid, rutin, luteolin, quercetin could bind stably with MAPK (-1,-3), AKT1, PRKC ( A, B).Conclusions Huoxue Jiedu Chinese Herbal Compatibility ( angelica sinensis -lonicerae japonicae flos) containing multiple active ingredients may play an anti-atherosclerosis role through the signaling pathways of MAPK, P13K-Akt and PRKC, reflecting the advantages of the treatment of activating blood circulation and detoxification of tra¬ditional Chinese medicine in anti-atherosclerosis.

doxorubicin induces chronic heart failure in rats by regulating mitochondrial dynamics via drp1/fundc1 pathway / 中国药理学通报

Aim To investigate the damage degree of doxorubicin hydrochloride( DOX )on cardiac function in rats, and to explore its possible mechanism. Methods Experiment 1: SD rats( n=48 )were randomly divided into control group( normal saline ), DOX 1 group( DOX cumulative dose 12 mg·kg-1 ;intraperitoneal injection ), DOX 2 group( 15 mg·kg-1 ;)and DOX 3 group( 18 mg·kg-1 ;). Cardiac structure and cardiac function were detected by echocardiography. B-type natriuretic peptide( BNP )was detected by ELISA. The morphological changes of myocardium were observed by Hematoxylin-eosin( HE )staining. The optimal dose group( DOX 2 group )was selected comprehensively. Experiment 2: SD rats( n=36 )were randomly divided into control group( normal saline ), DOX 2 group(15 mg·kg-1)and DOX 2+Mdivi-1 group( 15 mg·kg-1+daily abdominal injection of Mdivi-1(1 mg ·kg-1 ;)). Western blot was used to detect the protein expression of myocardial mitochondrial dynamics. Results Compared with the control group, hearts in DOX groups were enlarged and the heart function was reduced. Under the microscope, hypertrophy of cardiac cells and loose arrangement of cardiac fibers were observed in DOX group, and the higher the cumulative dose of DOX in rats, the more severe the degree of heart failure and the higher the mortality rate of rats. Compared with control group, the expression of mitochondrial dynamin-related protein 1( DRP1 )and related signaling pathway protein FUN14 domain containing 1( FUNDC1 )in DOX 2 group increased. The expression of optic atrophy 1( OPA1 )decreased, the expression of FUNDC1 and DRP1 protein decreased, and the expression of OPA1 protein was enhanced after the use of mitochondrial dynamics inhibitor(Mdivi-1). Conclusions DOX can cause chronic heart failure, and the mechanism may be related to DRP1/FUNDC1 mediated mitochondrial fission and fusion.

Effects of hsa_circ_0045943 targeting miR-106a on the biological characteristics of gastric cancer cells / 西安交通大学学报(医学版)

【Objective】 To investigate the effects of hsa_circ_0045943 targeting miR-106a on the biological characteristics of gastric cancer cells and its mechanism. 【Methods】 Human gastric cancer cells MKN-45， AGS and gastric mucosal epithelial cells GES-1 were cultured； circ_0045943 was detected by real-time polymerase chain reaction. The overexpression and silencing of circ_0045943 adenovirus vectors OE-circRNA and sh-circRNA together with their negative controls OE-NC and sh-NC were constructed and transfected； CCK-8 method was used to detect the proliferation activity of AGS cells after overexpression and silencing of circ_0045943； TUNEL method was used to detect the cell apoptosis； transwell assay was used to detect the cell migration and invasion； and would healing assay was used to detect the cell migration. Starbase database screened the binding site of miR-106a and circ_0045943. Real-time PCR was used to detect the expression of miR-106a， and the expression of circ_0045943 and the changes of miR-106a after the treatment of OE-circRNA and sh-circRNA. 【Results】 Real-time PCR showed that the expression of circ_0045943 decreased in gastric cancer cells MKN-45 and AGS compared to GES-1 （Pboth<0.001）. CCK-8 showed that the absorbance value of AGS cells in OE-circRNA group was lower than that in sh-circRNA group （P24 h<0.01， P48 h<0.001， and P72 h<0.001）. TUNEL showed the number of apoptotic AGS cells increased after overexpression of circ_0045943， but decreased after silencing of circ_0045943. Transwell assay showed that the migration and invasion of AGS cells were lower in OE-circRNA group than in sh-circRNA group （Pboth<0.001）. The wound healing assay showed that the migration rate of AGS cells in OE-circRNA group was the lowest， but was high in sh-circRNA group （P<0.001）. Starbase retrieved that circ_0045943 and miR-106a had complementary binding sequences. Real-time PCR showed that miR-106a was highly expressed in gastric cancer cells （P<0.001）， and the expressions of circ_0045943 and miR-106a significantly differed after treatment with OE-circRNA and sh-circRNA （Pboth<0.001）. With the increase or decrease of circ_0045943， the expression of miR-106a changed in the opposite direction. 【Conclusion】 Circ_0045943 has low expression in gastric cancer， and promoting or inhibiting circ_0045943 expression may regulate the proliferation， apoptosis， migration and invasion of gastric cancer cells by targeting miR-106a.

Clinical effect of different maintenance doses of caffeine citrate in the treatment of preterm infants requiring assisted ventilation: a pilot multicenter study / 中国当代儿科杂志

OBJECTIVES@#To explore the optimal maintenance dose of caffeine citrate for preterm infants requiring assisted ventilation and caffeine citrate treatment.@*METHODS@#A retrospective analysis was performed on the medical data of 566 preterm infants (gestational age ≤34 weeks) who were treated and required assisted ventilation and caffeine citrate treatment in the neonatal intensive care unit of 30 tertiary hospitals in Jiangsu Province of China between January 1 and December 31, 2019. The 405 preterm infants receiving high-dose (10 mg/kg per day) caffeine citrate after a loading dose of 20 mg/kg within 24 hours after birth were enrolled as the high-dose group. The 161 preterm infants receiving low-dose (5 mg/kg per day) caffeine citrate were enrolled as the low-dose group.@*RESULTS@#Compared with the low-dose group, the high-dose group had significant reductions in the need for high-concentration oxygen during assisted ventilation (P=0.044), the duration of oxygen inhalation after weaning from noninvasive ventilation (P<0.01), total oxygen inhalation time during hospitalization (P<0.01), the proportion of preterm infants requiring noninvasive ventilation again (P<0.01), the rate of use of pulmonary surfactant and budesonide (P<0.05), and the incidence rates of apnea and bronchopulmonary dysplasia (P<0.01), but the high-dose group had a significantly increased incidence rate of feeding intolerance (P=0.032). There were no significant differences between the two groups in the body weight change, the incidence rates of retinopathy of prematurity, intraventricular hemorrhage or necrotizing enterocolitis, the mortality rate, and the duration of caffeine use (P>0.05).@*CONCLUSIONS@#This pilot multicenter study shows that the high maintenance dose (10 mg/kg per day) is generally beneficial to preterm infants in China and does not increase the incidence rate of common adverse reactions. For the risk of feeding intolerance, further research is needed to eliminate the interference of confounding factors as far as possible.

Stasis Caused by Depression and Coronary Heart Disease / 中国实验方剂学杂志

Anxiety and depression are common comorbidities of coronary heart disease and are considered as independent risk factors in addition to traditional cardiovascular risk factors. Anxiety，depression and other mental abnormalities belong to the category of "depressive syndrome" of traditional Chinese medicine，which can lead to stasis of blood due to the lack of Qi flow. "Blood stasis" involves abnormal blood rheology， vascular endothelial dysfunction， chronic inflammatory response， abnormal lipid metabolism and other comprehensive pathological changes， and is the core pathogenesis of coronary heart disease in traditional Chinese medicine. "Depressive syndrome"can aggravate the development of coronary heart disease by promoting blood stasis in multiple ways. Prescriptions and herbs of promoting blood circulation and removing blood stasis can have a clinical effect by promoting blood circulation （improving physiological functions） and removing blood stasis （eliminating pathological changes）. In clinical practice， strengthening the screening of the mental and psychological status of patients with coronary heart disease and providing early and effective psychological interventions and combined Chinese and Western medicine drug treatment can significantly improve the clinical symptoms and prognosis of patients. This article was the first to put forward the academic view of "stasis caused by depression" for the first time，and discuss the modern biological research progress of "depression" in Chinese medicine that promotes blood stasis and aggravates coronary heart disease，in order to provide a basis for the subsequent development of Chinese medicine in the prevention and treatment of coronary heart disease. The aim is to provide a theoretical basis for the subsequent systematic research on the prevention and treatment of coronary heart disease with emotional abnormalities in Chinese medicine.

Evaluating the methodology of studies conducted during the global COVID-19 pandemic: A systematic review of randomized controlled trials / 中西医结合学报

BACKGROUND@#The therapeutic evidence collected from well-designed studies is needed to help manage the global pandemic of the coronavirus disease 2019 (COVID-19). Evaluating the quality of therapeutic data collected during this most recent pandemic is important for improving future clinical research under similar circumstances.@*OBJECTIVE@#To assess the methodological quality and variability in implementation of randomized controlled trials (RCTs) for treating COVID-19, and to analyze the support that should be provided to improve data collected during an urgent pandemic situation.@*SEARCH STRATEGY@#PubMed, Excerpta Medica Database, China National Knowledge Infrastructure, Wanfang, and Chongqing VIP, and the preprint repositories including Social Science Research Network and MedRxiv were systematically searched, up to September 30, 2020, using the keywords "coronavirus disease 2019 (COVID-19)," "2019 novel coronavirus (2019-nCoV)," "severe acute respiratory syndrome-related coronavirus-2 (SARS-CoV-2)," "novel coronavirus pneumonia (NCP)," "randomized controlled trial (RCT)" and "random."@*INCLUSION CRITERIA@#RCTs studying the treatment of COVID-19 were eligible for inclusion.@*DATA EXTRACTION AND ANALYSIS@#Screening of published RCTs for inclusion and data extraction were each conducted by two researchers. Analysis of general information on COVID-19 RCTs was done using descriptive statistics. Methodological quality was assessed using the risk-of-bias tools in the Cochrane Handbook for Systematic Reviews of Interventions (Version 5.1.0). Variability in implementation was assessed by comparing consistency between RCT reports and registration information.@*RESULTS@#A total of 5886 COVID-19 RCTs were identified. Eighty-one RCTs were finally included, of which, 45 had registration information. Methodological quality of the RTCs was not optimal due to deficiencies in five main domains: allocation concealment, blinding of participants and personnel, blinding of outcome assessment, incomplete outcome data, and selective reporting. Comparisons of consistency between published protocols and registration information showed that the 45 RCTs with registration information had common deviations in seven items: inclusion and exclusion criteria, sample size, outcomes, research sites of recruitment, interventions, and blinding.@*CONCLUSION@#The methodological quality of COVID-19 RCTs conducted in early to mid 2020 was consistently low and variability in implementation was common. More support for implementing high-quality methodology is needed to obtain the quality of therapeutic evidence needed to provide positive guidance for clinical care. We make an urgent appeal for accelerating the construction of a collaborative sharing platform and preparing multidisciplinary talent and professional teams to conduct excellent clinical research when faced with epidemic diseases of the future. Further, variability in RCT implementation should be clearly reported and interpreted to improve the utility of data resulting from those trials.

Analysis on Relationship Between Intestinal Flora and Diabetes Cognitive Impairment Based on "Paired Relationship Between Heart and Small Intestine" / 中国实验方剂学杂志

Based on theory of "paired relationship between the heart and the small intestine" in traditional Chinese medicine（TCM）， heart disease can affect the small intestine， and intestinal diseases can also affect the heart. The heart controls mental activities， so if the secretions function of small intestine is impaired， it will hinder the mind of the mind. Modern medicine has found that intestinal flora is closely related to cognitive function， which is consistent with the "paired relationship between heart and small intestine" in TCM. Diabetes cognitive impairment （DCI） is a category of "diabetes dementia" in TCM. According to TCM， the imbalance of spleen and stomach is the starting factor of diabetes dementia， and the phlegm turbidity of spleen deficiency is the pathological product of diabetes dementia. Previous projects have proved that intestinal flora disturbance is the modern pathological basis of spleen and stomach imbalance in TCM， suggesting that restoring spleen and stomach function to adjust intestinal microbial homeostasis is helpful for the prevention and treatment of diabetes dementia. At present， many achievements have been made in the study of the mutual causality between diabetes and intestinal flora disorder. In recent years， the incidence of DCI has increased rapidly， and the pathogenesis is complex and unclear， which increases the difficulty of treatment. Therefore， finding new targets should not be ignored. Given the complex connections among intestinal flora， diabetes and cognitive impairment， it is particularly important to tease out the relationship among the three. Based on previous scientific research， the author analyzed layer by layer and found that intestinal flora can not only affect the synthesis of neurotransmitters in the brain， but also participate in the common pathogenesis of diabetes and cognitive impairment such as chronic inflammation， insulin resistance and blood glucose rise， with a typical target effect. The pathogenic mechanism of intestinal flora in DCI was summarized in order to establish a new target and direction for the treatment， and to explore the scientific connotation of TCM in regulating spleen and stomach for treatment of diabetes dementia.

Comprehensive functional annotation of susceptibility variants identifies genetic heterogeneity between lung adenocarcinoma and squamous cell carcinoma / 医学前沿

Although genome-wide association studies have identified more than eighty genetic variants associated with non-small cell lung cancer (NSCLC) risk, biological mechanisms of these variants remain largely unknown. By integrating a large-scale genotype data of 15 581 lung adenocarcinoma (AD) cases, 8350 squamous cell carcinoma (SqCC) cases, and 27 355 controls, as well as multiple transcriptome and epigenomic databases, we conducted histology-specific meta-analyses and functional annotations of both reported and novel susceptibility variants. We identified 3064 credible risk variants for NSCLC, which were overrepresented in enhancer-like and promoter-like histone modification peaks as well as DNase I hypersensitive sites. Transcription factor enrichment analysis revealed that USF1 was AD-specific while CREB1 was SqCC-specific. Functional annotation and gene-based analysis implicated 894 target genes, including 274 specifics for AD and 123 for SqCC, which were overrepresented in somatic driver genes (ER = 1.95, P = 0.005). Pathway enrichment analysis and Gene-Set Enrichment Analysis revealed that AD genes were primarily involved in immune-related pathways, while SqCC genes were homologous recombination deficiency related. Our results illustrate the molecular basis of both well-studied and new susceptibility loci of NSCLC, providing not only novel insights into the genetic heterogeneity between AD and SqCC but also a set of plausible gene targets for post-GWAS functional experiments.

Efficacy and safety of modified chitosan eye drops in the treatment of Candida albicanskeratitis in rabbits / 中华实验眼科杂志

Objective:To evaluate the clinical efficacy and safety of modified chitosan eye drops on rabbit Candida albicans keratitis model.Methods:Ten healthy female New Zealand rabbits were used to establish the superficial Candida albicans keratitis model by the corneal surface lens method in the right eye.Slit lamp microscopy and corneal scraping and microscopic examination were performed to preliminarily determine whether the keratitis model has been successfully established, the rabbits were then randomly divided into a model group and a modified chitosan group by the random number table method.The successfully established rabbit models which were determined by fungal culture results were retained.Five normal rabbits receiving no intervention served as a normal control group.The experimental eyes in the normal control and modified chitosan groups were treated with modified chitosan eye drops, Six times a day for one week, and subsequently four times a day for one week.No treatment was administered to the model group.The changes of corneal lesions and ocular surfaces were examined by slit lamp microscopy every day.At 1, 7, 14, 21 and 28 days after modeling, the eye condition and corneal clinical scores was assessed by slit lamp microscopy.The corneal conditions in each group was observed for two weeks after drug withdrawal.Results:The corneal scraping and microscopic examination results of eight rabbits models showed that the fungal hyphae and spores were positive.The fungal culture results showed that the separated pathogen was Candida albicans.The success rate of modeling was 80%(8/10). The clinical scores in the model group at 7, 14 and 21 days after modeling were 14.50±0.58, 6.25±0.50 and 2.50±0.58, respectively, and were significantly higher than 7.25±1.26, 2.75±0.50 and 1.25±0.50 in the modified chitosan group (all at P<0.05). In the model group, corneal edema was significantly aggravated, and the central white ulcer area was enlarged within seven days after modeling.Between 7 and 28 days after modeling, the corneal ulcer was gradually healed, while the central corneal scar and neovascularization were remained.The average healing time was (24.5±2.6)days.In the modified chitosan group, the corneal infiltration was significantly alleviated within seven days after modeling, and the fungal hyphae and spores of corneal scraping were negative on the 14th day after modeling.The average healing time in the modified chitosan group was (13.5±1.3)days, which was significantly shorter than that in the the model group (P<0.01). No recurrence of keratitis was observed in the modified chitosan group after two weeks of drug withdrawal.The cure rate was 100%.In the normal control group, the conjunctival hyperemia, corneal edema, and lesions were not observed during topical administration.Conclusion:The treatment with modified chitosan eye drop is effective in a rabbit superficial Candida albicans keratitis model, and have no obvious toxic effects on ocular tissues.

DNA barcoding of 4 species of cheyletid mites based on COI and 18S rRNA gene sequences / 中国血吸虫病防治杂志

Objective To analyze the sequences of mitochondrial cytochrome C oxidase subunit I gene (COI) and 18S ribosomal RNA gene (18S rRNA), so as to identify the feasible DNA barcodes for 4 species of cheyletid mites and improve the DNA barcoding database for cheyletid mites. Methods Cheyletid mite samples were collected from small-scale flour mills in Fuyang, Wuhu and Tongling cities of Anhui Province from May 2018 to July 2019, extracted and morphologically identified. Then, genomic DNA was extracted from a single cheyletid mite, and the COI and 18S rRNA gene sequences were obtained by PCR amplification, cloning and sequencing. The obtained sequences were aligned using the BLAST software. Multiple sequence alignment was done using the software ClustalX version 1.83 using the known gene sequences from cheyletid mites. The genetic distance was calculated using the software MEGA X, and the phylogenetic tree was created using the maximum likelihood method. Results The DNA barcoding results of Cheyletus malaccensis, C. carnifex and Cheletomorpha lepidopterorum were consistent with the morphological identification, while no sequences pertaining to Eucheyletia reticulate were retrieved in the GenBank database. The proportions of A + T were 69.6% and 55.1% in the COI and 18S rRNA sequences of 4 cheyletid mites species, respectively, and the numbers of base substitutions were 137 and 46, respectively. There were 154 to 321 and 58 to 99 inter-species variation loci in the COI and 18S rRNA gene sequences of 4 cheyletid mites species, respectively, and the intra-species genetic distance was all 0.020 or less in the COI and 18S rRNA gene sequences of 4 cheyletid mites species, with inter-species genetic distance of 0.235 to 0.583 and 0.078 to 0.114, respectively. Phylogenetic analysis based on COI and 18S rRNA genes showed that all four species of cheyletid mites were clustered into a branch with a 100% supportive rate, which was consistent with the morphological identification. Conclusion Mitochondrial COI gene is superior to 18S rRNA gene as DNA barcodes for 4 species of cheyletid mites, which is more suitable to be used to investigate the phylogenetic relationship of at genus and species levels.

Genetic characteristics of hemagglutinin gene of H9N2 avian influenza viruses isolated from live poultry markets in Anhui Province from 2013 to 2018 / 中华疾病控制杂志

@#Objective To analyze genetic characteristics of hemagglutinin( HA) gene of H9N2 avian influenza viruses( AIVs) circulating in Anhui Province from 2013 to 2018． Methods All H9N2 positive samples tested by real－time polymerase chain reaction( PCＲ) were inoculated into specific patho- gen free ( SPF) chicken eggs for isolation and purification． Viral ＲNA was reversely transcribed into cD- NA and then amplified with gene specific primers． PCＲ products were sequenced and the gene sequences were analyzed using molecular and bioinformatics software． The DATAMONKEY online server was conducted to analyze selection pressure，and protein structure homology modelling was computed by the SWISS－MODEL server． Ｒesults 33 H9N2 AIVs isolated from live poultry markets belonged to h9．4．2．5 in the phylogenetic tree． The receptor binding sites of HA gene at 183，226 and 227 position were mutated into N，L and M，respectively． Meanwhile 189 and 190 sites presented with genetic polymorphism． Since 2015，all H9N2 viruses in this study carried 6 potential N－linked glycosylation sites． It was found that po- sition 160 of HA gene was subjected greater positive selection pressure，presented 7 spatial conformations at least． Conclusions The H9N2 viruses isolated from live poultry markets in Anhui Province possess the molecular characteristics of infecting mammals and the ability of antigenic drift，so we need to pay more attention to the genetic characteristics of the viruses．

Effect of miR-106a expression negatively regulated by KLF4 on the invasive activity of human gastric cancer cells BGC-823 / 西安交通大学学报(医学版)

Objective: To investigate the effect of Krüppel-like factor 4 (KLF4) on the expression of miR-106a and their interaction on the invasive activity of human gastric cancer cells. Methods: The JASPAR database was used to screen the transcriptional factor combined with miR-106a promoter. KLF4 over-expression vector KLF4-pcDNA and miR-106a reporter gene pmiR-106a-WT/MUT-luc were both constructed, and the dual luciferase reporter assay was used to detect the effect of KLF4 on the activity of miR-106a promoter. Forty specimens of human gastric cancer and their adjacent formalin-fixed paraffin-embedded (FFPE) specimens were collected, and Real-time PCR and immunohistochemistry were both used to detect the expression of KLF4. Human gastric cancer cell line BGC-823 was divided into four groups: miR-106a mimic, mimic NC, KLF4+pcDNA, and pcDNA basic. Transwell assay was employed to detect the invasive ability of the gastric cancer cells after four different treatments. Results: Dual luciferase reporter assay indicated that KLF4-pcDNA could antagonize the activity of miR-106a promoter, which suggested that the activity of pmiR-106a-WT-luc luciferase was inhibited (pmiR-106a-WT+pcDNA-basic compared with pmiR-106a-WT+pcDNA-KLF4, P0.05). FFPE samples showed that the relative expression of KLF4 in gastric cancer was 0.69±0.59, which significantly differed from that in adjacent paracancerous tissues (P<0.001). The expression of KLF4 was correlated with differentiation degree, lymph node metastasis, and infiltration depth of human gastric cancer (P<0.05). The positive expression of KLF4 was mainly located in the nucleus and cytoplasm of gastric mucosal epithelial cells. Transwell essay showed that the number of invasive cells in the four different treatment groups was not entirely the same (P<0.001): 89.00±14.85 for miR-106a mimic, 50.90±17.94 for mimic NC, 37.70±12.60 for KLF4+pcDNA, and 66.20±3.19 for pcDNA basic. Conclusion: Transcription factor KLF4 at least has direct binding with miR-106a promoter in vitro, which may be involved in the invasion and metastasis process of gastric cancer cell by negatively regulating the expression of miR-106a at the upstream transcription level.

Neuroprotective Effect of Tetrahydroxy Stilbene Glucoside on Aβ25-35-induced Neuron Synapses via CREB/BDNF Signaling Pathway / 中国实验方剂学杂志

Objective: To investigate the neuroprotective effect and mechanism of tetrahydroxy stilbene glucoside (TSG) on β-amyloid protein 25-35 (Aβ25-35)-induced neuron synapses damage. Method: Primary neurons were isolated and purified from cerebral cortex of suckling mouse. Then neurons were divided into control group, model group (incubation with Aβ25-35) and TSG groups (after incubation with Aβ25-35, add 6.25, 12.5, 25, 50, 100 μmol·L-1 TSG). Cell counting kit-8 (CCK-8) and Lactate dehydrogenase (LDH) methods were used to observe the viability of neuron, immunocytochemical staining was performed to determine the expressions of synapsin-1 (SYN-1), and the concentration of postsynaptic density-95 (PSD-95) and synaptophysin (SYP) were detected by enzyme-linked immunosorbent assay (ELISA) method. The level of cyclic adenosine monophosphate response element binding protein (CREB) and brain-derived neurotrophic factor (BDNF) mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR) and the level of CREB, Phosphorylated CREB (p-CREB) and BDNF proteins were determined by immunocytochemical staining or Western blot (WB). Result: Compared with normal group, the cell survival rate of model group was significantly reduced, LDH release was significantly increased (PPPPPPP-1,25 μmol·L-1 TSG can significantly enhance the expression of SYN-1(PPPPConclusion: TSG possesses the neuroprotective effect on Aβ25-35-induced neuron synapses, the mechanism may be associated with the activation of CREB/BDNF signaling pathway.

Clinical study of radical cystectomy under totally laparoscopy and ρ shape orthotopic ileal neobladder / 中华泌尿外科杂志

Objective@#To explore the feasibility and advantages of totally laparoscopic radical cystectomy plus ρshape orthotopic ileal neobladder, and to summarize the experiences.@*Methods@#The clinical data of 11 patients with bladder cancer admitted to the second hospital of Hebei medical university from October 2018 to May 2019 were retrospectively analyzed. All patients were male, aged 33 to 77 years, with an average of 64.4 years. Body mass index ranged from 18.0 to 31.8 kg/m2, with an average of 23.2 kg/m2. One case underwent partial cystectomy and 10 cases underwent transurethral resection of bladder tumor, with 10 cases of invasive urothelial carcinoma and 1 case of adenocarcinoma. No case underwent neoadjuvant therapy, and all cases were clinically staged as cT2a-3bN0-2M0. Totally laparoscopic radical cystectomy and ρ shape orthotopic ileal neobladder intraperitonealy were performed. The specific procedures were described as follows. After bladder resection, the ileal segment of 55 cm in length was intercepted at 25 cm from ileocecal valve, with the proximal part retaining 15 cm lumen as the input loop, and the distal part of 40 cm ileum being folded in 1∶1 ratio. The folded intestinal segment was made into a allantoic sac by using a linear incision closure device, forming a "ρ" shape with the input loop, bilateral ureters and the input loop. Anastomosis of wall, distal end of allantoic and urethra was performed.Record the perioperative data such as operation time, estimated bleeding, postoperative recovery, complications, and follow-up results.@*Result@#All of the 11 cases underwent successfully operation, and no cases were transferred to open surgery. The operation time ranged from 320 to 440 minutes, with an average of 357.1 minutes. The estimated amount of bleeding ranged from 100 to 300 ml, with an average of 207.1 ml. The total time of intestinal tract procedure was 80-100 minutes, with an average of 89.3 minutes, and the time of allantoic preparation was 14-19 minutes, with an average of 16.1 minutes. The pain score was 2-5 points at 4 hours after operation, with an average of 3.8 points, and 1-4 points at 24 hours after operation, with an average of 2.3 points. Postoperative exhaust time ranged from 2.5 to 3.5 days, with an average of 3.0 days.Catheter removed 21 days after operation, with 9 cases of urinary incontinence, including mild in 6 cases, moderate in 2 cases and severe in 1 case, with daytime pad of 0-3 and nighttime pad of 1-3.which improved gradually following pelvic exercise for 4-18 weeks. Postoperative hospital stay ranged from 7 to 13 days, with an average of 10.4 days. The drainage removal time was 4-11 days, with an average of 6.7 days. Postoperative pathology revealed 7 cases of high-grade invasive urothelial carcinoma, 3 case of low-grade invasive urothelial carcinoma and 1 case of adenocarcinoma. The tumors invade the prostate in adenocarcinoma patient, with left (3/13) and right (1/9) positive lymph nodes. One case of high-grade invasive urothelial carcinoma had both left (2/11) and right (1/9) positive lymph nodes, and the other cases were negative. The margin were negative in all patients. Pathological staging was pT2a-4aN0-2M0. Postoperative adjuvant chemotherapy with gemcitabine+ cisplatin regimen were performed in 9 patients.The follow-up period ranged from 3 to 29 weeks, with an average of 17.4 weeks, the patients with adenocarcinoma died of multiple organ failure at the 13th week after operation, and the other cases have no recurrence or metastasis. Dual J-tube was removed in 9 cases in the last follow-up, and the new bladder volume was estimated 300-350 ml, with residual urine of 0-43 ml and 19 ml of average. There is no stone formation in the new bladder.No hydronephrosis or ureteral dilatation aggravated.@*Conclusions@#Totally laparoscopic radical cystectomy plus ρ shape orthotopic ileal neobladder simplify the procedure of making allantoic storage and shorten the procedure time. The patients suffered less pain and recovered quickly after operation.It is a safe and feasible surgical procedure based on this study.

Clinical study of radical cystectomy under totally laparoscopy and p shape orthotopic ileal neobladder / 中华泌尿外科杂志

Objective To explore the feasibility and advantages of totally laparoscopic radical cystectomy plus ρshape orthotopic ileal neobladder,and to summarize the experiences.Methods The clinical data of 11 patients with bladder cancer admitted to the second hospital of Hebei medical university from October 2018 to May 2019 were retrospectively analyzed.All patients were male,aged 33 to 77 years,with an average of 64.4 years.Body mass index ranged from 18.0 to 31.8 kg/m2,with an average of 23.2 kg/m2.One case underwent partial cystectomy and 10 cases underwent transurethral resection of bladder tumor,with 10 cases of invasive urothelial carcinoma and 1 case of adenocarcinoma.No case underwent neoadjuvant therapy,and all cases were clinically staged as cT2a-3b N0-2 M0.Totally laparoscopic radical cystectomy and ρ shape orthotopic ileal neobladder intraperitonealy were performed.The specific procedures were described as follows.After bladder resection,the ileal segment of 55 cm in length was intercepted at 25 cm from ileocecal valve,with the proximal part retaining 15 cm lumen as the input loop,and the distal part of 40 cm ileum being folded in 1∶1 ratio.The folded intestinal segment was made into a allantoic sac by using a linear incision closure device,forming a “ρ” shape with the input loop,bilateral ureters and the input loop.Anastomosis of wall,distal end of allantoic and urethra was performed.Record the perioperative data such as operation time,estimated bleeding,postoperative recovery,complications,and follow-up results.Result All of the 11 cases underwent successfully operation,and no cases were transferred to open surgery.The operation time ranged from 320 to 440 minutes,with an average of 357.1 minutes.The estimated amount of bleeding ranged from 100 to 300 ml,with an average of 207.1 ml.The total time of intestinal tract procedure was 80-100 minutes,with an average of 89.3 minutes,and the time of allantoic preparation was 14-19 minutes,with an average of 16.1 minutes.The pain score was 2-5 points at 4 hours after operation,with an average of 3.8 points,and 1-4 points at 24 hours after operation,with an average of 2.3 points.Postoperative exhaust time ranged from 2.5 to 3.5 days,with an average of 3.0 days.Catheter removed 21 days after operation,with 9 cases of urinary incontinence,including mild in 6 cases,moderate in 2 cases and severe in 1 case,with daytime pad of 0-3 and nighttime pad of 1-3.which improved gradually following pelvic exercise for 4-18 weeks.Postoperative hospital stay ranged from 7 to 13 days,with an average of 10.4 days.The drainage removal time was 4-11 days,with an average of 6.7 days.Postoperative pathology revealed 7 cases of high-grade invasive urothelial carcinoma,3 case of low-grade invasive urothelial carcinoma and 1 case of adenocarcinoma.The tumors invade the prostate in adenocarcinoma patient,with left (3/13) and right (1/9) positive lymph nodes.One case of high-grade invasive urothelial carcinoma had both left (2/11) and right (1/9) positive lymph nodes,and the other cases were negative.The margin were negative in all patients.Pathological staging was pT2a-4a N0-2 M0.Postoperative adjuvant chemotherapy with gemcitabine + cisplatin regimen were performed in 9 patients.The follow-up period ranged from 3 to 29 weeks,with an average of 17.4 weeks,the patients with adenocarcinoma died of multiple organ failure at the 13th week after operation,and the other cases have no recurrence or metastasis.Dual J-tube was removed in 9 cases in the last follow-up,and the new bladder volume was estimated 300-350 ml,with residual urine of 0-43 ml and 19 ml of average.There is no stone formation in the new bladder.No hydronephrosis or ureteral dilatation aggravated.Conclusions Totally laparoscopic radical cystectomy plus ρ shape orthotopic ileal neobladder simplify the procedure of making allantoic storage and shorten the procedure time.The patients suffered less pain and recovered quickly after operation.It is a safe and feasible surgical procedure based on this study.

Epidemiological characteristics and repeated infection analysis of scrub typhus in Anhui Province from 2008 to 2017 / 中华疾病控制杂志

@# Objective To understand the epidemiological characteristics of scrub typhus in Anhui Province from 2008 to 2017, so as to provide scientific evidence for the development of control strategies and control measures. Methods Descriptive epidemiological methods were used to describe and analyze the cases of scrub typhus in Anhui Province from 2008 to 2017. Results The overall incidence of scrub typhus in Anhui Province showed an upward trend from 2008 to 2017 ( χ2=3 522.49, P<0.001). The highest incidence rate in 2016 was 3.98/100 000. The cases were concentrated from September to November. It was a typical “autumn and winter type” and peaked in October. It had the phenomenon of “high incidence in October”; the cases were mainly concentrated in the age group of 40-79 years old, accounting for 76.64% of the total reported cases. The cumulative number of cases in the 60-69 age group was the largest, and the incidence was mainly farmers. Conclusions In recent years, the incidence of scrub typhus in Anhui Province has shown an upward trend. It is necessary to strengthen the routine monitoring of scrub typhus in key areas and the detection of scrub typhus pathogens, and actively carry out prevention and health education of scrub typhus.