AIM: To delineate the specific mutation responsible for retinitis pigmentosa(RP)in a Yi pedigree, and to analyze the correlation of RHO gene mutation with clinical phenotype.METHODS:A comprehensive clinical evaluation was conducted on the proband diagnosed with RP and other familial members, complemented by a thorough ophthalmic examination. Peripheral blood samples were obtained from the proband and familial members, from which genomic DNA was extracte. Subsequent whole exome sequencing(WES)was employed to identify the variant genes in the proband. The identified variant gene was validated through Sanger sequencing, then an in-depth analysis of the mutation genes was carried out using genetic databases to ascertain the pathogenic mutation sites. Furthermore, an exhaustive analysis was performed to delineate the genotype and phenotype characteristics.RESULTS:The RP pedigree encompasses 5 generations with 42 members, including 19 males and 23 females. A total of 13 cases of RP were identified, consisting of 4 males and 9 females, which conforms to the autosomal dominant inheritance pattern. The clinical features of this family include an early onset age, rapid progression, and a more severe condition. The patients were found to have night blindness around 6 years old, representing the earliest reported case of night blindness in RP families. The retina was manifested by progressive osteocytoid pigmentation of the fundus, a reduced visual field, and significantly decreased or even vanished a and b amplitudes of ERG. The combined results of WES and Sanger sequencing indicated that the proband had a heterozygous missense mutation of the RHO gene c.1040C>T:p.P347L, where the 1 040 base C of cDNA was replaced by T, causing codon 347 to encode leucine instead of proline. Interestingly, this mutation has not been reported in the Chinese population.CONCLUSION:This study confirmed that the mutant gene of RP in a Yi nationality pedigree was RHO(c.1040C>T). This variant leads to the change of codon 347 from encoding proline to encoding leucine, resulting in a severe clinical phenotype among family members. This study provides a certain molecular, clinical, and genetic basis for genetic counseling and gene diagnosis of RHO.

AIM: To delineate the specific mutation responsible for retinitis pigmentosa(RP)in a Yi pedigree, and to analyze the correlation of RHO gene mutation with clinical phenotype.METHODS:A comprehensive clinical evaluation was conducted on the proband diagnosed with RP and other familial members, complemented by a thorough ophthalmic examination. Peripheral blood samples were obtained from the proband and familial members, from which genomic DNA was extracte. Subsequent whole exome sequencing(WES)was employed to identify the variant genes in the proband. The identified variant gene was validated through Sanger sequencing, then an in-depth analysis of the mutation genes was carried out using genetic databases to ascertain the pathogenic mutation sites. Furthermore, an exhaustive analysis was performed to delineate the genotype and phenotype characteristics.RESULTS:The RP pedigree encompasses 5 generations with 42 members, including 19 males and 23 females. A total of 13 cases of RP were identified, consisting of 4 males and 9 females, which conforms to the autosomal dominant inheritance pattern. The clinical features of this family include an early onset age, rapid progression, and a more severe condition. The patients were found to have night blindness around 6 years old, representing the earliest reported case of night blindness in RP families. The retina was manifested by progressive osteocytoid pigmentation of the fundus, a reduced visual field, and significantly decreased or even vanished a and b amplitudes of ERG. The combined results of WES and Sanger sequencing indicated that the proband had a heterozygous missense mutation of the RHO gene c.1040C>T:p.P347L, where the 1 040 base C of cDNA was replaced by T, causing codon 347 to encode leucine instead of proline. Interestingly, this mutation has not been reported in the Chinese population.CONCLUSION:This study confirmed that the mutant gene of RP in a Yi nationality pedigree was RHO(c.1040C>T). This variant leads to the change of codon 347 from encoding proline to encoding leucine, resulting in a severe clinical phenotype among family members. This study provides a certain molecular, clinical, and genetic basis for genetic counseling and gene diagnosis of RHO.

[摘 要] 目的：探讨溶瘤新城疫病毒（NDV）对IL-6诱导的人胶质母细胞瘤U87MG细胞增殖、迁移和侵袭的作用及其可能的机制。方法：将U87MG细胞分为对照组、IL-6组、NDV组、NDV+IL-6组，其中IL-6组与NDV+IL-6组用75 ng/mL IL-6预处理1 h，其余组用DMEM预处理1 h，后分别用DMEM、75 ng/mL IL-6、1 HU NDV、1 HU NDV+75 ng/mL IL-6处理24 h。MTT法、细胞划痕实验和Transwell侵袭实验分别检测IL-6、NDV对U87MG细胞增殖、迁移和侵袭的影响，WB法检测各组细胞JAK2、p-JAK2、STAT3、p-STAT3和MMP2蛋白的表达水平。结果：与对照组相比，IL-6组细胞迁移率显著升高（P<0.05），侵袭细胞数目显著增多（P<0.01）；与IL-6组相比，NDV+IL-6组U87MG细胞增殖率显著降低（P<0.05），细胞迁移率和侵袭细胞数目均显著降低（均P<0.01）。WB实验结果显示，与对照组相比，IL-6组p-STAT3/STAT3比值显著升高（P<0.01），NDV组p-JAK2/JAK2、p-STAT3/STAT3比值显著降低（P<0.05，P<0.01），MMP-2蛋白表达量显著降低（P<0.01）；与IL-6组相比，NDV+IL-6组p-STAT3/STAT3比值、MMP-2蛋白表达量均显著降低（均P<0.05）。结论：NDV能抑制IL-6对人脑胶质瘤U87MG细胞迁移和侵袭的诱导作用，其机制可能与JAK2/STAT3信号通路的参与调控有关。

ObjectiveThis study aimed to evaluate the clinical efficacy of Ruyi Zhenbaowan（RYZBW）in the treatment of initial and early knee osteoarthritis （KOA） through a prospective multicenter，randomized，double-blind，and placebo-parallel controlled trial. MethodFrom October 13^th， 2021 to December 25^th， 2021， 240 KOA subjects meeting the acceptance criteria were enrolled in 15 sub-centers including Wangjing Hospital， Chinese Academy of Chinese Medical Sciences， and they were randomly divided into observation group and control group， with 120 cases in each group. The intervention measures for the observation group were RYZBW + health education， and the intervention measures for the control group were RYZBW placebo + health education. The intervention period in both groups was four weeks， and they were followed up for four weeks after the intervention. The primary outcome measure was the total score of Western Ontario and McMaster University Osteoarthritis Index score （WOMAC score）， and the secondary outcome measures were the response rate of visual scale （VAS） pain score， WOMAC sub item scores （joint pain， joint stiffness， and joint function）， quality of life （SF-12） score， and traditional Chinese medicine （TCM） syndrome score. Result（1） Efficacy evaluation. The marginal model results showed that the observation group was better than the control group in improving the WOMAC total score and WOMAC pain score in the treatment of KOA with RYZBW， and the difference was statistically significant （P<0.05）. There was no significant difference between the two groups in improving VAS score response rate， WOMAC function score， WOMAC stiffness score， SF12-PCS （quality of life-physical health） score， SF12-MCS （quality of life-mental health） score， and TCM syndrome score. （2） Subgroup analysis. ① In terms of VAS score response rate， the response rate of the observation group was higher than that of the control group for subjects with baseline VAS score of （4， 5］， and the difference was statistically significant （P<0.05）. ② In terms of TCM syndrome score， for subjects aged ［56， 60］ and ［61， 65］， the decrease in total TCM syndrome score in the observation group was better than that in the control group， and the difference was statistically significant （P<0.05）. ConclusionTibetan medicine RYZBW has good clinical efficacy in improving WOMAC total score， VAS score response rate， WOMAC pain score， WOMAC function score， and TCM syndrome score for patients with initial and early KOA， which can fill the lack of Tibetan medicine RYZBW in the treatment of KOA and make a demonstration study for the inheritance and development of ethnic medicine.

Objective:To recheck the Histoplasma capsulatum and Coccidioides sp. strains of the past by molecular identification and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS). Methods:The phylogenetic relationships among the ten Histoplasma capsulatum isolates and reference strains were analyzed by multilocus sequence typing (MLST). Based on the Coi region, Coccidioides posadasii was distinguished from Coccidioides immitis accurately. MALDI-TOF MS was used to set up the MALDI-TOF MS database of Histoplasma capsulatum and Coccidioides sp. for rapid identification. In addition, hierarchical clustering of spectra was compared with MLST. Results:An unrooted dendrogram constructed with MLST showed that ten individuals of Histoplasma capsulatum were divided into three clades: Eurasia clade, Australia clade and North American class 2 clade, in agreement with the establishment by MALDI-TOF MS cluster analysis. All individuals of Coccidioides sp. were identified as Coccidioides posadasii with Coi region primers. The in-house MALDI-TOF MS database of Histoplasma capsulatum and Coccidioides posadasii was expanded and reached an identified accuracy of 100%. Conclusions:We improve the recognition of Histoplasma capsulatum and Coccidioides posadasii by molecular pathways which shows the major species or clades in Chinese mainland. The in-house MALDI-TOF MS database of Histoplasma capsulatum and Coccidioides posadasii can provide a new efficient way to identify those pathogens rapidly.

Objective To establish a fluorescent assay for rapid detection of Plasmodium falciparum based on recombinaseaided amplification (RAA) and CRISPR-Cas12a system,and to preliminarily evaluate the diagnostic efficiency of this system.. Methods The 18S ribosomal RNA (rRNA) gene of P. falciparum was selected as the target sequence, and three pairs of RAA primers and CRISPR-derived RNA (crRNA) were designed and synthesized. The optimal combination of RAA primers and crRNA was screened and the reaction conditions of the system were optimized to create a fluorescent RAA/CRISPR-Cas12a system. The plasmid containing 18S rRNA gene of the P. falciparum strain 3D7 was generated, and diluted into concentrations of 1 000, 100, 10, 1 copy/μL for the fluorescent RAA/CRISPR-Cas12a assay, and its sensitivity was evaluated. The genomic DNA from P. vivax, P. malariae, P. ovum, hepatitis B virus, human immunodeficiency virus and Treponema pallidum was employed as templates for the fluorescent RAA/CRISPR-Cas12a assay, and its specificity was evaluated. Fifty malaria clinical samples were subjected to the fluorescent RAA/CRISPR-Cas12a assay and nested PCR assay, and the consistency between two assays was compared. In addition, P. falciparum strain 3D7 was cultured in vitro. Then, the culture was diluted into blood samples with parasite densities of 1 000, 500, 200, 50, 10 parasites/μL with healthy volunteers’ O-positive red blood cells for the RAA/CRISPR-Cas12a assay, and the detection efficiency was tested. Results The Pf-F3/Pf-R3/crRNA2 combination, 2.5 μL as the addition amount of B buffer, 40 min as the RAA reaction time, 37 °C as the reaction temperature of the CRISPR-Cas12a system were employed to establish the fluorescent RAA/CRISPR-Cas12a system. Such a system was effective to detect the plasmid containing 18S rRNA gene of the P. falciparum strain 3D7 at a concentration of 1 copy/μL, and presented fluorescent signals for detection of P. falciparum, but failed to detect P. ovum, P. malariae, P. vivax, T. pallidum, hepatitis B virus or human immunodeficiency virus. The fluorescent RAA/CRISPR-Cas12a system and nested PCR assay showed completely consistent results for detection of 50 malaria clinical samples (kappa = 1.0, P < 0.001). Following 6-day in vitro culture of the P. falciparum strain 3D7, 10 mL cultures were generated and the fluorescent RAA/CRISPR-Cas12a system showed the minimal detection limit of 50 parasites/μL. Conclusion The fluorescent RAA/CRISPR-Cas12a system is rapid, sensitive and specific for detection of P. falciparum, which shows promising value for rapid detection and risk monitoring of P. falciparum.

Objective:To analyze the prevalence of different genotypes of wild measles virus in Fujian province from 2014 to 2019.Methods:The information of suspected measles cases and the throat swab specimens of the cases in Fujian province from 2014 to 2019 were collected. Reverse transcription-polymerase chain reaction was applied to amplify the 450 nucleotides of the 3-terminal of the nucleoprotein gene. Phylogenetic tree was constructed to identify and analyze the target sequence.Results:A total of 1 102 suspected measles cases were reported in Fujian province from 2014 to 2019, with an annual incidence rate of 0.47/100 000. Totally 884 throat swabs were tested in the laboratory, and 179 nucleotide sequences were obtained except vaccine strain. Sequence analysis showed that 110 measles genotypes were H1, 43 were B3 and 26 were D8. Nucleotide homology analysis showed that the H1a genotype measles virus in Fujian province distributed in two branches, divided into nine different sequence variants. Some of the variants were highly homologous (99.8%-100.0%) with the measles strains in other regions and countries. There were six different sequence variants of B3 genotype measles virus in Fujian province. The nucleotide homology between the B3 genotype measles virus in Fujian province and the domestic and foreign prevalent B3 strains was 98.4%-100.0%. The D8 genotype measles strain in Fujian province had five different sequence variants, some of which were highly homologous with the strains from Vietnam, Japan and Thailand.Conclusions:Three different genotypes of measles viruses (H1, B3 and D8) prevailed in Fujian province from 2014 to 2019, among which H1 genotype was still the local endemic genotype, while B3 and D8 genotypes were imported genotypes in Fujian province. The result indicated that although the local cases of measles were less in Fujian province in recent years, the risk of imported cases from abroad was increasing, and the surveillance of imported cases from abroad should be strengthened.

Objective:To summarize and compare the characteristics of oral microbiota in women during the preconception period and the third trimester.Methods:This retrospective cohort study involved 55 women who were recruited in the Preconceptional Offspring Trajectory Study (PLOTS) conducted by Fudan University and followed up to the third trimester in the Maternal and Child Health Care Hospital of Jiading District of Shanghai from September 2016 to December 2019. A total of 110 unstimulated saliva samples were collected in the preconception period ( n=55) and the third trimester ( n=55). Features of oral microbiota in the samples were analyzed by 16S rRNA gene-based sequencing. Moreover, the related factors were also analyzed. Paired t test or Wilcoxon matched-pairs signed-ranks test were used to analyze the differences in α-diversity during preconception and the third trimester; t test, analysis of variance (ANOVA), Kruskal-Wallis test and Mann-Whitney U test for comparison between groups with different characteristics and permutational multivariate analysis of variance (PerMANOVA) for β-diversity were used; Linear discriminant analysis (LDA) effect size (LEfSe 1.0) was used to identify the iconic oral flora. Results:(1) The Ace index of oral microbiota was significantly lower in the third trimester than that in the preconception period [661.14(578.15-752.85) vs 730.64 (632.40-911.00), T=1 077.00, P=0.010]. There was also a significance difference in β-diversity ( F=12.539, R2=0.104, P=0.001). Some species such as Saccharibacteria_TM7_G3, Prevotella_7, Absconditabacteria_SR1_G1, Porphyromonas, Ruminococcaceae_UCG_014, Prevotella, Peptostreptococcus, Prevotella_2, Alloprevotella, Parvimonas, Solobacterium and Eubacterium_nodatum_group in saliva were statistically more abundant in the third trimester than those in the preconception period (all P<0.05). (2) The third-trimester Shannon index was lower among those with lower income [5.44 (5.08-5.77) vs 5.75 (5.44-6.12), U=219.00, P=0.029] and those with gargle habit after meal or dessert [5.36 (4.91-5.48) vs 5.72 (5.44-6.05), U=374.00, P=0.046]. Conclusions:The features of oral microbiota vary in women during the preconception period and the third trimester. There is a significant increase in the abundance of oral pathogenic and opportunistic bacteria in the third trimester.

Objective:To evaluate the clinical effects of painting therapy on fatigue, anxiety and depression in patients with hematological neoplasms undergoing chemotherapy.Methods:Totally 78 patients with hematological neoplasms admitted to the laminar flow ward in a tertiary hospital in Shanghai from October 2018 to May 2019 were enrolled. Patients who were included from October 2018 to January 2019 constituted the control group (39 cases), others from February to May 2019 constituted the intervention group (39 cases). Patients in both groups received routine health education and psychological care, and the intervention group received extra 4 weeks of painting intervention once a week. The Revised Piper Fatigue Scale (PFS-R), Self-rating Anxiety Scale (SAS) and Self-rating Depression Scale (SDS) were used to assess fatigue and emotional status of two groups during chemotherapy before the intervention, after the intervention and one month after the intervention. One case in control group was lost to follow-up while three cases in intervention group.Results:At 2 times points after painting therapy, total fatigue、anxiety and depression score in the intervention group were (4.04±0.91), (48.81±3.92), (46.06±4.53), (3.55±0.84), (47.47±4.33), (45.17±4.74) points; those in the control group were (5.34±1.18), (51.13±3.88), (50.63±4.36), (5.07±1.40), (52.74±4.04), (51.24±4.01) points.The time effect on fatigue of each dimension and total scores, anxiety and depression scores was significant ( F values were 48.859-126.096, P<0.05); the group effect and the interaction effect of intervention and time on sensory, emotional, behavioral and total fatigue scores, anxiety and depression scores were significant ( F values were 6.037-37.014, P<0.05; F values were 7.696-35.754, P<0.05). Conclusion:Painting therapy can effectively alleviate fatigue and relieve anxiety and depression in patients with hematological neoplasms undergoing chemotherapy.

OBJECTIVE：To understand the situation of healthcar e workers’occupational exposure of hazardous drugs （chemotherapeutic drugs and antiviral drug ）and relative cognition level ，and to provide reference for improving the level of occupational protection. METHODS ：During Oct. 2018 to Mar. 2019，healthcare workers from 12 hospitals of different levels and different departments in Shaanxi province were selected as respondents to conduct a self-designed questionnaire survey. Information we surveyed contained baseline characteristics ，hazardous drugs exposure ，physical health status ，and occupational protection. Valid questionnaires were collected and analyzed statistically. RESULTS ：A totally of 1 848 questionnaires were sent out ，and 1 767 questionnaires were collected ，including 95 were pharmacists ，100 were physicians ，and 1 572 were nurses. The frequency of diarrhea and menstrual disorders in healthcare workers with long-term exposure to antiviral drugs （antiviral drug exposure group ） and cross-exposure to antiviral drugs and chemotherapeutic drugs （cross-exposure group ）were significantly higher than unexposed group （P＜0.05）. The incidence of routine blood abnormalities in cross-exposure group ，antiviral drug exposure group and healthcare workers with long-term exposure to chemotherapy drugs （chemotherapy drug exposure group ） was higher than unexposed group by 9.13％，5.50％ and 12.34％，respectively. 84.7％ of the respondents had little knowledge of hazardous drugs ， and 8.15％ of the respondents had never received occupational protection training. Additionally ，the ratio of healthcare workers receiving occupational protection training in the cross-exposure group was significantly higher than unexposed group （P＜0.05）， and the ratio of healthcare workers receiving occupational protection training in antiviral drug exposure group were significantly lower than unexposed group （P＜0.05）. CONCLUSIONS ：Long-term exposure to hazardous drugs will cause certain occupational hazards to healthcare workers. It is necessary to improve healthcare workers ’awareness of self-protection ，increase the input of hospitals in occupational protection training and establish occupational protection standards in order to improve the current situation of occupational protection of healthcare workers.