Abnormalities, Multiple ; genetics ; Acidosis, Renal Tubular ; genetics ; pathology ; therapy ; Arthrogryposis ; genetics ; pathology ; therapy ; Biopsy ; Carrier Proteins ; genetics ; Cholestasis ; genetics ; pathology ; therapy ; DNA Mutational Analysis ; Humans ; Infant, Newborn ; Mutation ; Syndrome ; Vesicular Transport Proteins ; genetics

Humans ; Lymphoma ; diagnosis ; therapy ; Molecular Targeted Therapy

Abstract: Objective To investigate the molecular characteristics of the H9N2 avian influenza virus (AIV) causing human infection in Yunnan Province in 2019, and to provide the scientific basis for the prevention and control of avian influenza in Yunnan Province. Methods Influenza virus typing was performed by real-time RT-PCR in two influenza-like illness samples, and the Illumina Miseq high-throughput sequencer was used to determine the viral genome sequence. HA and NA gene sequence alignment and phylogenetic tree construction were performed using Mega7.0 software. Results Real-time RT-PCR results showed that two influenza-like illness samples were positive for H9N2 subtype. The full length of HA and NA were obtained by genomic sequencing. Sequence system evolution analysis showed that the HA and NA of the two AIVs in Yunnan Province were in the same evolutionary clade as A/Chicken/Zhejiang/HJ/2007 and belonged to the G57 type. The HA nucleotide and amino acid homology of the two AIVs were 93.92% and 95.00%, respectively, and the NA nucleotide and amino acid homology was 93.31% and 82.03%, respectively. The nucleotide (amino acid) homology of HA was 92.29%-96.94% (93.77%-98.43%) and 92.84%-94.92% (94.18%-96.23%), respectively, and NA nucleotide homology (amino acid) were 91.81%-97.60% (77.82%-94.83%), 94.38%-97.22% (85.47%-94.55%), respectively, compared with that of human infected H9N2 epidemic strains obtained in China from 2015 to 2020. Both AIVs HA protein cleavage site sequences were PSRSSR↓GLF, which was in line with the characteristics of low pathogenic influenza. The analysis of HA protein receptor binding site showed that amino acids at positions 109, 161, 163, 191, 202, 203 and 234 were consistent with the reference strains, while amino acids at position 198 were mutated to T. N166D and 168N mutations were also found in HA protein, and both AIVs had 7 potential glycosylation sites. Analysis of the erythrocyte binding site of NA gene found that there were amino acid mutations at positions 369, 402, 403, and 432, and amino acid deletion at positions 63-65 was found in the NA genes. There were 4 and 5 potential glycosylation sites in the two AIVs, respectively, and no drug resistance site mutations were found. Conclusions　The receptor binding sites, erythrocyte binding sites and glycosylation sites of HA and NA genes of H9N2 AIV in Yunnan Province have different degrees of variation, and monitoring and prevention and control should be strengthened.

Background and purpose:Five-year survival rate of post-operation patients with non-small cell lung cancer(NSCLC)is less than 40%.Treatments after recurrence are difficult.Our study aimed to evaluate the efficacy of pemetrexed on postoperative recurrence of NSCLC.Methods:From Jan.2006 to Sep.2008,40 NSCLC with postoperative recurrence were observed.All patients had received pemetrexed(ALIMTA)500 mg/m2 or carboplatin eonbined.Results:Among the 40 patients,partial response in 10 patients(25.00%),stable disease in 19 patients(47.50%),progressive disease in 11 patients(27.50%).The total response rate was 25.00%and clinical benefit control rate was 72.50%.Pemetrexed had significantly better disease control rate in female than in male (9 1.30% vs 47.06%,P=0.034),in adenocarcinoma patients than in non-adenocarcinoma's(87.10% vs 22.22%,P=0.001).Median overall survival time(MST)was 10.70 months.Progression-free survival time(PFS)was 5.18 months.Adenocarcinoma patients had longer PFS than non-adenocarcinoma patients.Conclusion:Pemetrexed demonstrates significant antitumor activity and good tolerance in these patients.

Objective:To evaluate the effect of left ventricular hypertrophy and deformation on cardiac function in patients with uremic cardiomyopathy (UCM) by using the technology of two dimensional speckle tracking imaging (2D-STI).Methods:A total of 67 UCM patients were randomly divided into the normal cardiac function group (subgroup A,32 cases) and the abnormal cardiac function group (subgroup B,35 cases)according to the New York Heart Association points (NYHA-P).A total of 30 healthy subjetcs served as the control group.Parameters including left ventricular ejection fraction (LVEF),left ventricular mass index (LVMI),left ventricular spherical index (LVSI),left ventricular myocardial mean radial strain (MRS),mean radial strain rate (MRSR),mean longitudinal strain (MLS),local systolic twist angle (STA),and mitral annulus maximum displacement (TMAD) were detected.Results:MLS,MRS,MRSR,LVSI,STA and TMAD in the Group A and Group B were lower than that in the control group (P＜0.05),and LVMI in the Group A and Group B was increased than those in the control group (P＜0.05);LVEE MLS,MRS,MRSR,LVSI and STAin the Group B was decreased than that in the Group A (P＜0.05).MLS in the Group A and B were positively correlated with LVEF and LVSI,but negatively correlated with LVMI.Using the point of 14.10％ for MLS to evaluate UCM patients with NYHA-P＞4 points,the sensitivity,the specificity and Yuedden index were 90.5％,71％ and 0.585,respectively.STA in UCM patients were lower than that in the control (P＜0.05).Conclusion:2D-STI possesses a unique advantage in detecting left ventricular strain and strain rate on left ventricular regional function in UCM with left ventricular hypertrophy and ventricular deformation.There is no direct correlation between the left ventricular hypertrophy and ventricular deformation,but the ventricular hypertrophy and deformation are correlated with regional cardiac function and clinical cardiac function.Left ventricular regional dysfunction may occur before cardiac hypertrophy and deformation.

Objective To analyze the survival status, distribution characteristics and social security of newly diagnosed occupational pneumoconiosis patients in Guizhou Province from 2006 to 2021. Methods The newly diagnosed pneumoconiosis cases reported in Guizhou Province from 2006 to 2021 were collected from the “Occupational Diseases and Hazards Monitoring Information System” under “National Health Insurance Disease Prevention and Control Information System”. Telephone or face-to-face surveys were conducted to investigate these patients. Results A total of 12 413 newly diagnosed pneumoconiosis patients were reported in Guizhou Province from 2006 to 2021, with 11 192 cases included in the follow-up, and 10 631 cases were followed-up successfully. According to the follow-up study, 10 565 cases (accounting for 99.4%) were survived, and 66 cases (accounting for 0.6%) died. The number of pneumoconiosis showed an increasing trend followed by a sharp decline from 2006 to 2021, reaching its peak in 2016. The main industry of the newly pneumoconiosis cases was concentrated in the mining industry (accounting for 90.6%). The top three regions with surviving cases in the follow-up were Bijie City, Zunyi City, and Qiannan Prefecture, accounting for 34.0%, 25.4% and 12.0%, respectively. The main types of pneumoconiosis were coal workers' pneumoconiosis and silicosis, accounting for 67.1% and 30.8%, respectively. Most cases were in the age group of 50 to <60 years old, accounting for 59.1%, and the majority of the workers had worked in dusty environments for 5 to <25 years, accounting for 91.5%. In terms of social security, about 89.5% of cases were in the basic medical insurance for urban and rural residents which was the most popular social security. The employer's compensation rate was 67.7%, and the work-related injury insurance participation rate was 51.5%. Conclusion Pneumoconiosis cases in Guizhou Province exhibit significant regional disparities. It has a high concentration in the industry, a younger age profile, and limited social security coverage. It is necessary to strengthen the special management of dust and industry supervision in mining industry, intensify follow-up work for pneumoconiosis cases, reinforce the construction of pneumoconiosis rehabilitation station in key regions, and improve the quality of life of pneumoconiosis cases.

OBJECTIVETo investigate promoter methylation status of LITAF gene in B-cell lymphoma and to explore transcription regulation of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on LITAF gene.

METHODSOne hundred and five paraffin specimens including 54 cases of diffuse large B cell lymphoma (DLBCL), 15 small lymphocytic lymphoma (SLL), 8 mucosa-associated lymphoid tissue lymphoma (MALT) and 6 follicular lymphoma (FL) were included. Five reactive lymphoid hyperplasia samples were collected as control. Methylation status of CpG island in LITAF gene in the specimens and in Raji, Pfeiffer and Daudi cell lines were detected by methylation-specific PCR (MSP). LITAF expression in Raji, Pfeiffer and Daudi cell lines with or without 5-Aza-CdR treatment was detected by Western blot and immunohistochemistry. The inhibitory ratio in the three cell lines was measured by MTT assay.

RESULTSThe frequency of LITAF gene methylation in B-cell lymphoma was 89.5% (94/105) . Among them, 3.8% (4/105) showed complete hypermethylation. In control group, however, there was no methylation in CpG island of LITAF gene promoter. The expression of LITAF was recovered or increased along with the cell growth inhibition when the cells exposed to demethylating reagent.

CONCLUSIONSLITAF gene silencing with aberrant CpG methylation is probably one of the critical events to the oncogenesis of B-cell lymphoma, which may have important implications as a candidate marker for diagnosis and target gene therapy.

Adult ; Azacitidine ; analogs & derivatives ; metabolism ; Cell Line, Tumor ; CpG Islands ; DNA Methylation ; Gene Silencing ; Humans ; Lymphoma, B-Cell ; genetics ; metabolism ; Lymphoma, B-Cell, Marginal Zone ; genetics ; Lymphoma, Large B-Cell, Diffuse ; genetics ; Nuclear Proteins ; genetics ; metabolism ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Transcription Factors ; genetics ; metabolism

Objective:To investigate the effect of Enolase inhibition (ENOblock) on autophagy- related protein expression and motor function promotion after spinal cord injury in rats.Methods:A total of 160 female SD rats were divided into sham-operation group, 3-methyladenine (3-MA) autophagy inhibitor treatment group (3-MA group), spinal cord injury group and ENOblock treatment group (ENOblock group) according to the random number table, with 40 rats per group. Back laminectomy without injury to the spinal cord was performed in sham-operation group. Spinal cord injury at T 8 was induced by using a modified Allen weight-drop apparatus to establish a spinal cord injury model in the rest three groups. 3-MA and ENOblock groups were injected 3-MA (2.5 mg/kg) and ENOblock (100 μg/kg) into the caudal vein immediately after injury, respectively. Sham-operation and spinal cord injury groups were injected same dose of isotonic sodium chloride solution into the caudal vein. At 1, 3, 7, 14 and 21 days after injury, BBB score was used to evaluate lower limb motor function. At day 3 after injury, the ratio of microtubule-associated protein 1 light chain 3 (LC3)-II to LC3-I and protein expressions of autophagy effector protein (Beclin-1) and polyubiq-uitinbinding protein (p62) were detected by Western blotting. At day 7 after injury, LC3-Ⅱ and Beclin-1 positive cells in the injured area of the spinal cord were determined by immunofluorescence staining. At day 3 after injury, the mRNA expressions of Beclin-1 and Enolase in the injured area of the spinal cord were detected by RT-PCR. Results:At 1, 3, 7, 14 and 21 days after injury, BBB score was lowered in 3-MA group [(1.4±1.1)points, (2.4±0.9)points, (3.8±1.8)points, (7.6±1.1)points, (9.0±2.1)points], spinal cord injury group [(0.8±0.5)points, (1.8±0.9)points, (3.6±0.9)points, (6.2±1.3)points, (8.0±0.7)points] and ENOblock group [(2.0±0.9)points, (2.2±0.8)points, (4.8±1.1)points, (10.6±1.5)points, (13.2±0.8)points] compared to sham-operation group [(21.0±0.0)points at all time points] (all P<0.05). Moreover, the score in ENOblock group was significantly higher than that in spinal cord injury group at 14, 21 days after injury, and the score in 3-MA group was significantly higher than that in spinal cord injury group at day 21 after injury (all P<0.05). At day 3 after injury, Western blotting showed that the ratio of LC3-II to LC3-I and protein expressions of Beclin-1 and p62 were 0.46±0.10, 0.41±0.03, 0.81±0.03 in sham-operation group, 0.66±0.06, 0.69±0.02, 0.59±0.05 in 3-MA group, 0.85±0.06, 1.07±0.03, 0.41±0.02 in spinal cord injury group and 0.68±0.06, 0.66±0.08, 0.55±0.02 in ENOblock group. By comparison, spinal cord injury group showed significantly higher ratio of LC3-II to LC3-I and protein expression of Beclin-1 and significantly lower protein expression of p62 than sham-operation group (all P<0.05); 3-MA and ENOblock groups showed significantly lower ratio of LC3-II to LC3-I and protein expression of Beclin-1 and significantly higher protein expression of p62 than spinal cord injury group (all P<0.05); there was no significant difference in the ratio of LC3-II to LC3-I and protein expressions of Beclin-1 and p62 between 3-MA and ENOblock groups (all P>0.05). At day 7 after injury, immunofluorescence staining showed that LC3-II and Beclin-1 positive cells in 3-MA and ENOblock groups were less than those in spinal cord injury group. At day 3 after injury, RT-PCR showed that mRNA expressions of Beclin-1 and Enolase in spinal cord injury group (1.08±0.16, 0.98±0.17) were higher than those in sham-operation group (0.25±0.06, 0.29±0.03). Moreover, mRNA expressions of Beclin-1 and Enolase in 3-MA group (0.77±0.11, 0.72±0.04) and ENOblock group (0.81±0.10, 0.64±0.09) were lower than those in spinal cord injury group (all P<0.05). There was no significant difference in mRNA expressions of Beclin-1 and Enolase between 3-MA and ENOblock groups (all P>0.05). Conclusions:Autophagy activity is significantly up-regulated after spinal cord injury in rats. ENOblock can inhibit autophagy and promote motor function recovery in rats by regulating the expression of autophagy-related proteins.

Objective To discover the medicinal active molecules from the fermentation extract of sponge-symbiotic Streptomyces sp. LHW2432. Methods Compounds were isolated and purified from the fermentation extract of LHW2432 by silica gel, ODS chromatographic columns, and HPLC. The structures of the compounds were elucidated based on the analyses of modern spectrum technologies and the related literatures research. Through plate coating method and broth microdilution method, the antimicrobial activities were tested by the indicator strains of Bacillus mycoides, methicillin-resistant Staphylococcus aureus (MRSA), Mycobacterium smegmatis, Candida Albicans, and Escherichia coli. Results Five compounds were discovered and their structures were identified as descycloavandulyl-lavanduquinocin (1), N-acetyltyramine (2), phomapyrone C (3), germicidin A (4), and germicidin I (5). Compound 1 showed inhibitory activities against MRSA (MIC, 100 μg/ml) and M. smegmatis (MIC, 64 μg/ml), respectively. Conclusion Five compounds were discovered from LHW2432, among which compound 1 was a new natural product and could be used as a precursor of the tricyclic carbazole alkaloids with neuroprotective activity. Moreover, compound 1 showed weak inhibitory activities against gram-positive pathogenic bacteria.

The polar tube protein is the major component of polar tube, and can specifically locate on the polar tube of microsporidia and plays an important role in invasion host cell. In this study, we analyzed the potential O- and Nglycosylation sites in polar tube protein 1 from Nosema bombycis. NbPTP1 was successfully cloned to eukaryotic expression vector pMT/Bip/V5-His A, involved V5 and His tags. After transfection, NbPTP1 gene could be efficiently expressed in Drosophila S2 cells. In addition, Lectin blotting and beta elimination analysis showed that NbPTP1 expressed in Drosophila S2 cells was O-glycosylation. These studies provided a basis for understanding the relationship between glycosylation and function of NbPTP1, helped us to reveal the infection mechanism of microsporidia and established effective diagnosis and prevention methods for microsporidia.