Fragile X syndrome(FXS)is caused by abnormal duplication and amplification of the FMR1 gene CGG.This article reports a pair of brothers diagnosed with FXS by genetic testing.Two patients,aged 15 and 14 years old respectively,both had clinical manifestations such as language disorders,intellectual disabilities,attention deficit disorder,autism spectrum disorder,and FXS's characteristic facial features.The proband had a rare late-onset epileptic seizure,which was well treated with levetiracetam,while his younger brother had no electroencephalogram abnormalities after repeated follow-up.This pair of cases suggests that the clinical phenotype of FXS has diversity and heterogeneity.

Objective:To investigate the differential gene expression profiling between subcutaneous fat (SF) and orbital septum fat (OF).Methods:Six samples of SF tissue were collected from patients undergoing abdomen or thigh liposuction, and six samples of OF tissue were collected from patients undergoing double eyelid or pouch surgery in Chongqing Xing Rong Plastic Surgery Hospital from May 2022 to June 2022. Transcriptional characteristics of SF and OF were compared by RNA sequencing. The gene expression differences between SF and OF were analyzed by bioinformatics methods. The differences in gene expression were verified by real-time quantitative PCR (qRT-PCR).Results:Principal component analysis (PCA) showed the SF and OF groups were obviously separated as two independent gene clusters. The volcano plot analysis displayed the upregulated genes (such as FOXL2 and FOXL2NB), or the downregulated genes (such as DEFB132 and HOXB3) in SF group compared to those in OF group. Besides, the protein-protein interaction (PPI) network revealed the top 3 upregulated hub genes in SF group (MYH7, TCAP and MYL2) and in OF group (SOX9, WNT1 and WNT2). Heatmap analysis based on the above PPI showed that the two groups of upregulated genes were clustered and separated obviously. In addition, the signaling pathway analysis revealed that cardiomyopathy, muscle contraction and AMPK pathway were enriched in the SF group, while Wnt singling and cancer-related pathway were enriched in OF group. Finally, the qRT-PCR showed that SOX9 ( P<0.001) and WNT2 ( P<0.001) were upregulated, while DEFB132 ( P<0.001) and HOXB3 ( P<0.01) were downregulated in OF group compared to SF group, respectively. Conclusions:Compared to SF group, SOX9 and WNT2 are regulated, while DEFB132 and HOXB3 are downregulated in OF group. The differentially expressed genes in OF may be the key factors mediating its unique physiological features.

Objective To knock out cdc42 gene in zebrafish using CRISPR/Cas9 technology,and investigate the effect of cdc42 on early osteochondral development.Methods After the conservation of cdc42 gene sequence of different species was analyzed by multiple sequence alignment analysis,guide RNA of cdc42 gene was designed,and cdc42 knockout zebrafish was constructed by CRISPR/Cas9 technology.The expression pattern of cdc42 was detected by whole-mount in situ hybridization,and the chondrogenesis phenotype was observed by transgenic labeled fish line Tg(col2a1a:GFP),and vertebral mineralization was observed by alizarin red staining.Results Multiple sequence alignment analysis showed that cdc42 was highly conserved in human,mouse and zebrafish,and in situ hybridization results showed that cdc42 was expressed in a variety of tissues in the head and whole body,including mandibular cartilage.With the aid of guide RNA of cdc42,cdc42 knockout zebrafish was successfully constructed by CRISPR/Cas9 technology.The cdc42 mutants exhibited shortened body length(P＜0.01)and delayed cranial development at 3 d post fertilization,with small heads and eyes(P＜0.01),as well as delayed mandibular development.The Tg(col2a1a:GFP)zebrafish showed that the mutants presented abnormal morphology of Meckel's cartilage and ceratohyal cartilage cells,with disordered arrangement of chondrocytes and increased angle and decreased length in ceratohyal cartilage(P＜0.01).The homozygous mutants died at 10～13 d after fertilization.The results of alizarin red staining suggested delayed vertebral mineralization and reduced endochondral ossification of the mutants.Conclusion CRISPR/Cas9 technology successfully knocks out the cdc42 gene in zebrafish,resulting in delayed development of jaw cartilage,delayed mineralization of the vertebrae,and decreased endochondral ossification.

Objective To enhance the triage efficiency of patients with acute heart failure(AHF)by improving emergency severity index(ESI)score,and to achieve stratified care for patients with AHF.Additionally,the aim is to improve the combination of stratified scores and nursing care in emergency departments to address the limitations of current studies.Methods A retrospective nested cohort study was conducted on 120 patients with AHF admitted to the Affiliated Hospital of Guizhou Medical University from January 1,2019 to December 30,2021.According to the clinical outcomes of the patients,the patients were divided into subgroup A(71 cases,recovered after treatment,no serious complications during treatment),subgroup B[35 cases,discharged after treatment,multiple organ dysfunction syndrome(MODS),acute respiratory distress syndrome(ARDS),shock and other serious complications occurred during treatment],and subgroup C(14 cases,cardiac death during treatment).A prospective randomized controlled study involving 106 patients with AHF admitted to our hospital from January 1 to December 30,2022,was conducted,and the patients were divided into control group(53 cases)and stratified care group(53 cases)according to the random number table method.The control group received routine nursing care,while patients in the stratified care group received stratified care based on the ESI scoring clinical care pathway.The early warning score model of elderly patients with AHF was constructed with risk threshold as warning signal.The improved ESI scoring criteria were used to score AHF patients,with a total score of 25 points.The higher the score,the higher the risk.0-10 is divided into Ⅰ level risk,>10-20 is divided into Ⅱ level risk,>20 is divided into Ⅲ level risk.Implement nuring at grades Ⅲ,Ⅱ,and Ⅰ,respectively.The changes of modified ESI scores in subgroup A,subgroup B and subgroup C were observed,and Spearman correlation coefficient was used to evaluate the correlation between modified ESI scores and clinical outcomes of different severity of disease.Receiver operator characteristic curve(ROC curve)was drawn to evaluate the effectiveness of the risk threshold classification method of different nursing plan designs in the stratified nursing group and the control group,and to compare the efficiency differences of emergency nursing rescue time,rescue success rate,and stay time in the emergency department,and the incidence of complications such as MODS,ARDS,shock and nursing satisfaction during treatment between the stratified nursing group and the control group.Results The modified ESI score in subgroup C was the highest,significantly higher than that in subgroup B and A(19.6±1.7 vs.17.8±1.5,15.3±1.3,all P<0.05).Spearman correlation analysis showed that the modified ESI score was significantly positively correlated with the occurrence of complications,the occurrence of cardiac death,and the recovery after treatment(r values were 0.623,0.635,0.322,P values were 0.004,0.003,0.012,respectively).Therefore,an improved ESI score has a certain early warning effect on complications and cardiac death of patients.ROC curve analysis showed that the area under the ROC curve(AUC)of routine care for stratified care was 0.710 and 0.620,and the 95％ confidence interval(95％ CI)was 0.620-0.750 and 0.580-0.690,respectively,with Pvalues of 0.023 and 0.034.It shows that the difference between the two nursing methods is significant,and further indicates that the risk threshold classification method designed in this study is effective.With the increase of risk,the rescue time and emergency stay time of the two groups were gradually extended,and the success rate of rescue was gradually decreased,the rescue time and emergency stay time of grade Ⅲ risk were the longest and the success rate of rescue was the lowest,and the rescue time and emergency stay time of the stratified nursing group were significantly shorter than those of the control group(minutes:59.6±6.3 vs.76.5±7.2 and 57.6±5.4 vs.68.2±7.1,both P<0.05),the success rate of rescue was significantly increased[(68.7±6.1)％ vs.(54.6±5.2)％,P<0.05],and the difference of rescue time between grade Ⅰ risk and grade Ⅲ risk was the largest.There were significant differences in the success rate of rescue and the duration of emergency stay in grade Ⅲ risk.The incidence of complications such as MODS,ARDS and shock in stratified care group was significantly lower than that in control group[incidence of MODS:13.2％(7/53)vs.18.9％(10/53),incidence of ARDS:15.1％(8/53)vs.22.6％(12/53),incidence of shock:13.2％(7/53)vs.20.8％(11/53),all P<0.05],satisfaction of patients with service attitude,operation technique,comfort and timeliness were significantly improved compared with the control group[service attitude:28.3％(15/53)vs.18.9％(10/53),operation technology:30.2％(16/53)vs.20.8％(11/53),comfort:32.1％(17/53)vs.24.5％(13/53),timeliness:32.1％(17/53)vs.24.5％(13/53),all P<0.05].Conclusion Based on the modified ESI score,early risk warning thresholds are divided and stratified nursing is conducted,which is beneficial for timely intervention in AHF,effectively ensuring the clinical nursing value of patients during the implementation of medical measures,and has broad application prospects.

Background A large number of studies on fluoride-induced systemic bone damage have been reported previously, but there is little understanding of the characteristics of fluoride accumulation in jawbone. Jawbone is homologous to the other bone tissues in the body, and is an indispensable and important frame structure in the oral cavity. Objective To study fluoride accumulation and its change trends in teeth, jawbone, and femur of SD rats with chronic drinking-water-borne fluorosis. Methods A total of 144 three-week-old SD rats, half male and half female, were randomly divided into two groups, a normal control group and a fluoride group. The rats in the normal controlgroup drank purified water disinfected and filtered from Guizhou, and the water contained 0.08 mg·kg⁻¹ fluoride which was lower than the national water quality standard at 1 mg·kg⁻¹. The rats in the fluoride group were fed with sodium fluoride (NaF) solution with a concentration of 150 mg·L⁻¹. At 3, 5 and 7 months of the fluoride exposure, the levels of fluoride in urine, blood, teeth, jawbone, and femur were measured by fluoride ion electrode method. Results There was no sex difference in fluoride content in different biological samples of rats in the fluoride group and the normal control group (all P>0.05). After 3 months of fluoride exposure, the rats in the fluoride group showed dental fluorosis of grade II, and higher levels of fluoride ion in blood and urine than the normal control group (all P<0.05), indicating that the rat model of fluoride drinking-water-borne chronic fluorosis was successfully replicated. In the normal control group, the levels of fluoride in femur remained stable; at the end of 3 months, the levels of fluoride in jawbone and teeth were (1097.36±470.34) and (453.09±173.43) mg·kg⁻¹ respectively, and at the end of 7 months, the levels of fluoride in jawbone and teeth were (2113.18±634.49) and (1604.80±160.43) mg·kg⁻¹ respectively. Both jawbone and teeth showed a positive temporal effect of increasing fluoride accumulation (P<0.05). After continuous fluoride feeding, the fluoride levels in jawbone, teeth, and femur of rats in the fluoride group were (3145.02±765.82), (1550.20±77.73), and (3640.55±699.42) mg·kg⁻¹ after 3 months, and (8420.36±1728.56), (4702.08±1417.06), and (6091.99±1384.97) mg·kg⁻¹ after 7 months. The three kinds of hard tissues all showed a positive temporal effect of increasing fluoride accumulation (P<0.05), and the cumulative increas was large than that in the normal control group. Among them, jawbone fluorine increased most. At the end of 5 months, the levels of fluoride in jawbone, femur, and teeth were (6485.02±2141.98), (4914.99±1529.41), and (3365.21±1462.27) mg·kg⁻¹ respectively, and the levels of fluoride in jawbone was much higher than those in femur and teeth (P<0.05). Conclusion Hard tissues such as bones and teeth are fluorine sensitive tissues. Compared with femur, jawbone showed significantly high fluoride accumulation, while teeth show relatively lagging fluoride accumulation.

Objective:To observe the effect of fluoride on growth and development of bone microstructure of rats condyle subchondral bone (RCSB).Methods:Forty two 3-week-old SD rats (half male and half female) were fed adaptively for 1 week, and 3 females and 3 males were sacrificed and recorded as 0 month. The remaining rats were randomly divided into control group ( n = 18) and fluoride exposed group ( n = 18) according to their body weight (55 - 70 g), half male and half female. The fluoride exposed group was fed with 150 mg/L sodium fluoride (NaF) aqueous solution, and the control group was fed with tap water. The two groups of experimental animals were sacrificed at 3, 5 and 7 month, respectively, 6 rats in each group, half male and half female. The right mandibular condyle was separated, and Micro CT scanning was performed to detect the microstructure parameters of RCSB. Results:In fluoride exposed group (3 month), bone surface/tissue volume (BS/TV), bone surface/bone volume (BS/BV), trabecular thickness (Tb.Th), trabecular number (Tb.N), structure model index (SMI), connectivity, connectivity density (Conn.D) and total porosity of female rats were significantly different from those of male rats ( t = - 5.10, - 5.58, 4.52, - 4.32, - 4.03, - 2.81, - 6.71, - 3.32, P < 0.05). There was no significant difference in each index between female and male rats in fluoride exposed group (5, 7 month, P > 0.05). Conclusion:In chronic fluorine exposure bone environment, the RCSB bone microarchitecture of male and female rats is different with time, showing the tendency of fluoride injury that the bone changes of female rats are slowed and the bone changes of male rats are active.

Objective:The investigation and research on the application status of Hepatic Venous Pressure Gradient (HVPG) is very important to understand the real situation and future development of this technology in China.Methods:This study comprehensively investigated the basic situation of HVPG technology in China, including hospital distribution, hospital level, annual number of cases, catheters used, average cost, indications and existing problems.Results:According to the survey, there were 70 hospitals in China carrying out HVPG technology in 2021, distributed in 28 provinces (autonomous regions and municipalities directly under the central Government). A total of 4 398 cases of HVPG were performed in all the surveyed hospitals in 2021, of which 2 291 cases (52.1%) were tested by HVPG alone. The average cost of HVPG detection was (5 617.2±2 079.4) yuan. 96.3% of the teams completed HVPG detection with balloon method, and most of the teams used thrombectomy balloon catheter (80.3%).Conclusion:Through this investigation, the status of domestic clinical application of HVPG has been clarified, and it has been confirmed that many domestic medical institutions have mastered this technology, but it still needs to continue to promote and popularize HVPG technology in the future.

[摘 要] 目的：探讨长基因间非编码RNA（long intergene non-coding RNA，LINC）01018是否通过抑制miR-297调控胃癌HGC-27细胞的增殖、凋亡及放射敏感性。方法： 收集于青海省第五人民医院接受手术的胃癌患者（21例）及化疗抵抗胃癌患者（19例）的癌组织和癌旁组织，以qPCR检测胃癌组织、化疗抵抗胃癌组织和胃癌HGC-27细胞中LINC01018、miR-297的表达。双荧光素酶报告基因实验验证LINC01018与miR-297之间的靶向关系。在HGC-27细胞中转染LINC01018过表达质粒pcDNA-LINC01018或miR-297抑制剂，或共转染pcDNA-LINC01018和miR-297模拟物，以qPCR检测验证细胞转染效果。MTT和克隆形成实验检测转染后HGC-27细胞的增殖活力与克隆形成能力，流式细胞术检测转染后HGC-27细胞的凋亡率，克隆形成实验检测各组转染后HGC-27细胞的放射敏感性，WB实验检测细胞中Ki67、cleaved-caspase3、pro-caspase3蛋白的表达。结果： 与癌旁组织或胃正常黏膜上皮细胞GES-1相比，胃癌组织、化疗抵抗胃癌组织和胃癌HGC-27细胞中LINC01018呈低表达、miR-297呈高表达（均P<0.01）。LINC01018与miR-297存在靶向结合关系，LINC01018负向调控miR-297的表达。过表达LINC01018或敲减miR-297可抑制HGC-27细胞的增殖活力与克隆形成能力、降低细胞存活率，促进细胞的凋亡、下调Ki67和pro-caspase3蛋白表达水平、上调cleaved-caspase3蛋白表达水平、提高HGC-27细胞的放射敏感性（均P<0.01）。共转染pcDNA-LINC01018和miR-297模拟物可逆转过表达LINC01018对HGC-27细胞的所有上述作用，尤其可逆转其对HGC-27细胞的放射增敏作用（P<0.05或P<0.01）。结论： LINC01018通过下调miR-297表达抑制胃癌HGC-27细胞增殖而促进凋亡和增强细胞的放射敏感性，该作用与Ki67和caspase3的表达变化有关。

Objective: To explore the valuable indicators for differential diagnosis by comparing the clinical features of Crohn′s disease (CD) with primary intestinal lymphoma (PIL). Methods: From 2010 to 2017, at The Seventh Medical Center of PLA General Hospital, a total of 91 patients diagnosed with CD or PIL were enrolled, including 76 cases of CD, 14 cases of PIL and one case of CD with secondary lymphoma. The clinical data of enrolled patients were retrospectively analyzed. T-test, non-parametric test and chi-square test were used for statistical analysis. Results: The average age at diagnosis of CD patients was (37.7±16.0) years, which was younger than that of PIL patients (52.6±19.6) years, and the difference was statistically significant (t=-3.085, P=0.003). The median duration of CD was 36.0 months, which was longer than that of PIL (3.5 months), and the difference was statistically significant (Z=-3.616, P<0.01). Abdominal pain and extra-intestinal manifestations (oral ulcers, joint injuries, erythema nodosum and perianal lesions) were more common in CD patients, and the differences were all statistically significant (χ²=9.427 and 5.173, both P<0.05). CD patients were not diagnosed by colonoscopic biopsy alone. Totally 13 of 14 patients were diagnosed by colonoscopic biopsy and immunohistochemisty and one patient was diagnosed after surgery. The diagnostic rate of pathological biopsy in PIL group was significantly higher than that in CD group (χ²=82.584, P<0.01). One patient was initially clinically diagnosed as CD, and then developed secondary lymphoma after five times of infliximab treatment. Conclusions The diagnosis of CD is generally supported by young age, long course of disease, abdominal pain and extra-intestinal manifestations. The diagnostic rate of colonoscopic biospsy in PIL patients is higher than that in CD patients.

Purpose To explore the expression and significance of histone methyltransferase EZH2,clinicopathological features in primary testicular diffuse large B-cell lymphoma (DLBCL).Methods Immunohistochemical of Ventana Ultra View two-step staining was used to detect expression of EZH2 in 17 cases of primary testicular DLBCL.The relationship between EZH2 expression and its clinicopathological features were analyzed.Sanger squencing was used to detect EZH2 Y641 mutation in these cases.Results Morphologically,the tumor cells resembled centroblasts in 11 cases,immunoblasts in 3 cases,and anaplastic variants in 3 cases.Immunophenotypically,14 cases were non-germinal centre B cell like (non-GCB) type and 3 cases were germinal centre B cell like (GCB) subtype.EZH2 overexpressed in all 17 cases.EZH2 overexpressed in nearly tumor celts with uniformly strong intensity in 15 cases,and more than 70％ tumor cells with moderate to strong intensity in 2 cases.The follow-up information was obtained in 9 patients,with a median survival time of 35 months.No association was found between the level of EZH2 expression and outcome of patients.No mutation of EZH2 Y641 was detected.Conclusion Primary testicular DLBCL is a rare aggressive B cell lymphoma with distinctive clinicopathological features.Detection of EZH2 expected to assist in the diagnosis and treatment of tumor.