Effects of different intervention methods on psychological anxiety, stress, and fatigue among healthcare workers during COVID-19 pandemic: a systematic review and meta-analysis.

OBJECTIVE: This study aimed to explore the treatment effects of various intervention methods on the stress, anxiety, and fatigue of medical workers during the 2019 Coronavirus Disease (COVID-19) pandemic. MATERIALS AND METHODS: We conducted computer searches in both Chinese and English databases including China National Knowledge Infrastructure (CNKI), Wanfang Data Knowledge Service Platform (Wang Fang), VIP Chinese Journal Service Platform (VIP), Web of Science, Embase, PubMed, Cochrane Library, Scopus, and ScienceDirect to include prospective randomized controlled studies (Prospective RCT) published before September 30, 2023, regarding different treatment methods for stress, anxiety, and fatigue among healthcare workers during COVID-19. Data on anxiety, stress, and fatigue of research participants were extracted from the included studies, followed by statistical analysis of treatment effects using R software with the meta package. RESULTS: A total of 9 articles were eventually included, involving a total of 1,466 participants, including 686 in the control group and 780 in the study group. Intervention measures included mindfulness-based therapy in 4 studies and other intervention methods in 5. The anxiety status of the health workers was assessed using the Generalized Anxiety Disorder-7 (GAD-7), and the meta-analysis revealed a pooled mean difference (MD) of -0.53 (95% CI: -1.42, 0.37). Stress status was evaluated by Perceived Stress Scale 4 (PSS-4), and the meta-analysis results showed a post-treatment MD of 0.13 (95% CI: -0.39, 0.65), with a pre- and post-treatment difference MD of -0.44 (95% CI: -2.65, 1.76). Maslach Burnout Inventory (MBI) was employed for the evaluation of fatigue. The meta-analysis results showed an MD of -6.13 (95% CI: -16.68, 4.43) for the MBI Emotional index, an MD of 5.04 (95% CI: -3.25, 13.33) for the Personal Accomplishment index, and an MD of -1.68 (95% CI: -6.50, 3.13) for the Depersonalization index. CONCLUSIONS: Maintaining the mental health of frontline health workers is crucial during the COVID-19 pandemic, and mindfulness-based therapy is the most extensively employed psychological intervention method. However, its effectiveness requires further research confirmation.

[Determination of 4-methyl-2-pentanol in workplace air by solvent desorption-gas chromatography].

Objective: To establish a method for the determination of 4-methyl-2-pentanol in the air of workplace by gas chromatography. Methods: In January 2022, 4-methyl-2-pentanol in the air of workplace was collected by activated carbontube, eluted with dichloromethane-methanol (95∶5, V/V), separated by capillary column and determined by gas chromatogram. Results: The limit of detection for 4-methyl-2-pentanol was 0.04 µg/ml. The linear range of 4-methyl-2-pentanol was 0.16-1616.60 µg/ml, with the regression equation of y=1.94x-5.48, and the coefficient correlation was 0.99958, and the minimum detection concentration was 0.03 mg/m(3) (collected sample volume was 1.50 L). The within-run precisions were 1.08%-1.75% and the between-run precisions were 1.41%-2.52%. The desorption rates were 95.15%-99.91%. The samples could be stored at least 3 days at room temperature and 7 days at 4 ℃ without significant loss. Conclusion: The method has the advantages of good precision, high sensitivity and simple operation. It is suitable for the determination of 4-methyl-2-pentanol in the air of workplace.

[Determination of acetylacetone in workplace air by solvent desorption-gas chromatography].

Objective: To establish a method for the determination of acetylacetone in the air of workplace by gas chromatography. Methods: In August 2020, acetylacetone in the air of workplace was collected by silica gel tube, eluted with methanol, separated and detected by gas chromatography with flame ionization detector. The detection limit and precision of the method were also analyzed. Results: The linear range of acetylacetone was 1.95-1950.60 µg/ml with the regression equation of y=0.815x-3.667, and the correlation coefficient was 0.99993. The limit of detection of the method was 0.18 µg/ml and the minimum detection concentration was 0.12 mg/m(3) (collected sample volume was 1.50 L). The within-run precisions were 1.08%-4.11% and the between-run precisions were 1.98%-2.80%. The desorption rates were 99.68%-100.45%. The sealed samples could be kept at least 15 days at room temperature without significant loss. Conclusion: The solvent desorption-gas chromatography method for the determination of acetylacetone has good precision, high sensitivity and simple operation, and is suitable for the determination of acetylacetone in the air of the workplace.

[Screening, functional analysis and clinical validation of differentially expressed genes in diabetic foot ulcers].

Objective: To screen the differentially expressed genes (DEGs) in diabetic foot ulcers (DFUs), and to perform functional analysis and clinical validation of them, intending to lay a theoretical foundation for epigenetic therapy of chronic refractory wounds. Methods: An observational study was conducted. The gene expression profile dataset GSE80178 of DFU patients in Gene Expression Omnibus (GEO) was selected, and the DEG between three normal skin tissue samples and six DFU tissue samples in the dataset was analyzed and screened using the GEO2R tool. For the screened DEG, ClusterProfiler, org.Hs.eg.db, GOplot, and ggplot2 in the R language packages were used for Gene Ontology (GO) enrichment analysis of biological processes, molecular functions, and cellular components, and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, respectively. Protein-protein interaction (PPI) analysis was performed using STRING database to screen key genes in the DEG, and GO enrichment analysis of key genes was performed using Cytohubba plug-in in Cytoscape 3.9.1 software. DFU tissue and normal skin tissue discarded after surgery were collected respectively from 15 DFU patients (7 males and 8 females, aged 55-87 years) and 15 acute wound patients (6 males and 9 females, aged 8-52 years) who were admitted to Xiang'an Hospital of Xiamen University from September 2018 to March 2021. The mRNA and protein expressions of small proline-rich repeat protein 1A (SPRR1A) and late cornified envelope protein 3C (LCE3C) were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction and immunohistochemistry, respectively. Data were statistically analyzed with independent sample t test. Results: Compared with normal skin tissue, 492 statistically differentially expressed DEGs were screened from DFU tissue of DFU patients (corrected P<0.05 or corrected P<0.01), including 363 up-regulated DEGs and 129 down-regulated DEGs. GO terminology analysis showed that DEGs were significantly enriched in the aspects of skin development, keratinocyte (KC) differentiation, keratinization, epidermal development, and epidermal cell differentiation, etc. (corrected P values all <0.01). KEGG pathway analysis showed that DEGs were significantly enriched in the aspects of tumor-associated microRNA, Ras related protein 1 signaling pathway, and pluripotent stem cell regulatory signaling pathway, etc. (corrected P values all <0.01). PPI analysis showed that endophial protein, SPRR1A, SPRR1B, SPRR2B, SPRR2E, SPRR2F, LCE3C, LCE3E, keratin 16 (all down-regulated DEGs), and filoprotein (up-regulated DEG) were key genes of DEGs screened from DFU tissue of DFU patients, which were significantly enriched in GO terms of keratinization, KC differentiation, epidermal cell differentiation, skin development, epidermis development, and peptide cross-linking, etc. (corrected P values all <0.01). The mRNA expressions of SPRR1A and LCE3C in DFU tissue of DFU patients were 0.588±0.082 and 0.659±0.098, respectively, and the protein expressions were 0.22±0.05 and 0.24±0.04, respectively, which were significantly lower than 1.069±0.025 and 1.053±0.044 (with t values of 20.91 and 13.66, respectively, P values all <0.01) and 0.38±0.04 and 0.45±0.05 (with t values of 9.69 and 12.46, respectively, P values all <0.01) in normal skin tissue of acute wound patients. Conclusions: Compared with normal skin tissue, there is DEG profile in DFU tissue of DFU patients, with DEGs being significantly enriched in the aspects of KC differentiation and keratin function. Key DEGs are related to the biological function of KC, and their low expressions in DFU tissue of DFU patients may impede ulcer healing.

[The method of determination for butyronitrile and isobutyronitrile in the air of workplace by dissolved desorption-gas chromatography].

Objective: To establish a method for the determination of butyronitrile and isobutyronitrile in the air of workplace by gas chromatography. Methods: In March 2020, butyronitrile and isobutyronitrile in the air of workplace was collected by silica gel, eluted with methanol, separated and determined by gas chromatogram with flame ionization detector, the characteristics of determination of nitrile and isobutyronitrile by gas chromatography were analyzed. Results: The limit of detection for butyronitrile and isobutyronitrile was 0.33 µg/ml. The linear range of butyronitrile determined by this method was 1.60-1600.00 µg/ml, y=2.295x-3.480, and the coefficient correlation was 0.99998, and the minimum detection concentration was 0.22 mg/m(3) (collected sample volume was 1.50 L) . The within-run precisions were 2.43%-4.12%, the between-run precisions were 1.72%-3.70%, and the desorption rates were 93.26%-98.41%. The linear range of isobutyronitrile determined by this method was 1.52-1520.00 µg/ml, y=2.208x-0.102, and the coefficient correlation was 0.99998, and the minimum detection concentration was 0.22 mg/m(3) (collected sample volume was 1.50 L) . The within-run precisions were 2.52%-3.22%, the between-run precisions were 1.20%-3.82%, and the desorption rates were 96.85%-102.50%. The sealed samples could be stored at least 10 days at room temperature without significant loss. Conclusion: The method has the advantages of good precision, high sensitivity and simple operation. It is suitable for the simultaneous determination of butyronitrile and isobutyronitrile in the air of workplace.

[Clinical and microbiological characteristics of hypervirulent Klebsiella pneumoniae lung abscess].

Objective: To study the clinical and microbiological characteristics of hypervirulent Klebsiella pneumoniae (hvKP) lung abscess, and to compare with the classic Klebsiella pneumoniae (cKP) lung abscess. Methods: A total of 18 patients with Klebsiella pneumoniae lung abscesses admitted to the First Affiliated Hospital of Zhejiang University School of Medicine from January 2017 to September 2020 enrolled. The strains with positive result of string test were defined as hvKP, and the negative strains were defined as cKP. The patients' basic diseases, symptoms, laboratory data and other clinical characteristics were collected. The microbiological characteristics of the strains included as following: VITEK method to determine the in vitro susceptibility of the strains to antibiotics; PCR to detect the capsular serotypes and virulence genes. The differences in clinical characteristics and microbiological characteristics of strains between hvKP group and cKP group were compared. Results: Among the 18 patients with Klebsiella pneumoniae lung abscess, 12 were hvKP infection, mainly male (10 cases), with a median age of 59.0 years; 8 cases in the hvKP group had an onset time of ≤2 weeks, and the median onset time was 10.5 days. There were significantly more diabetes (12 cases) and extrapulmonary abscesses (11 cases) in hvKP group than those in cKP group (both P<0.001). The extrapulmonary abscesses in the hvKP group were mainly liver abscesses (10 cases), and 4 cases were multi-site (≥3) abscesses. The number of indwelling catheters and invasive procedures before infection were higher in cKP group than those in hvKP group (both P=0.025). The imaging of Klebsiella pneumoniae lung abscess was mainly subpleural with the size of less than 10 cm. There were more multiple abscesses cases in hvKP group (9 cases) than cKP group (P=0.009). The median interval time between the detection of a pulmonary abscess and an extrapulmonary abscess was 1.0 day. The resistance rate of common antibiotics was significantly lower in hvKP than cKP. Conclusions: hvKP lung abscesses are more common in patients with diabetes, and the clinical manifestations are nonspecific. The lung imaging manifestations are multiple subpleural abscesses, indicating hematogenous dissemination. Liver abscesses were present in most cases, suggesting the source of infection. The main virulent gene of hypervirulent Klebsiella pneumoniae is aero. For patients with hvKP lung abscess, attention should be paid to finding hidden lesions.

[Meta-analysis of ß-blockers for the primary prevention of liver cirrhosis with clinically significant portal hypertension with no or small esophageal varices].

Objective: To explore whether NSBB is suitable for the primary prevention of liver cirrhosis accompanied by CSPH with no or small esophageal varices. Methods: Relevant literatures were retrieved from Cochrane library, PubMed, EMBASE, SinoMed, CNKI and Wanfang databases until December 12, 2020. All randomized controlled trials (RCTs) on NSBB use for primary prevention of cirrhosis accompanied by CSPH with no or small esophageal varices were collected. The literature was strictly screened according to the established inclusion and exclusion criteria, odds ratio (OR), and 95% confidence interval (CI) combined effect size. The development of esophageal varices and the initial upper gastrointestinal bleeding were the primary outcome measures. Death (with a maximum average follow-up of about five years) and adverse events (adverse drug reactions, etc.) were the secondary outcome measures. Results: A total of 9 RCTs with 1396 cases were included. Meta-analysis results showed that, compared with placebo, NSBB significantly reduced the incidence of liver cirrhosis accompanied by CSPH with no or small esophageal varices to large esophageal varices progression (OR=0.51, 95%CI: 0.29-0.89, P=0.02), and mortality (with maximum average follow-up of about five years) (OR=0.64, 95%CI: 0.44-0.92, P=0.02); however, there was no statistically significant difference in the initial upper gastrointestinal bleeding rate between the two groups (OR=0.82, 95%CI: 0.44-1.52, P=0.53). Adverse event incidence was greater in the NSBB than the placebo group (OR=1.74, 95%CI: 1.27-2.37, P=0.0005). Conclusions: NSBB use cannot reduce the initial upper gastrointestinal bleeding rate or adverse event incidence in patients with liver cirrhosis accompanied by CSPH with no or small esophageal varices, but it can delay the progression of gastroesophageal varices and reduce patient mortality.

[Bacterial community diversity in Dermatophagoides farinae using high-throughput sequencing].

OBJECTIVE: To investigate the bacterial community diversity in Dermatophagoides farinae. METHODS: Laboratory-cultured D. farinae was collected, and the composition of microbial communities was determined by sequence analyses of the V4 region in the bacterial 16S ribosomal RNA (16S rRNA) gene on an Illumina PE250 high-throughput sequencing platform. Following quality control and filtering of the raw sequence files, valid reads were obtained and subjected to operational taxonomic units (OTU) clustering and analysis of the composition of microbial communities and alpha diversity index using the Usearch software, Silva database, and Mothur software. RESULTS: A total of 187 616 valid reads were obtained, and 469 OTUs were clustered based on a sequence similarity of more than 97%. OTU annotation showed that the bacteria in D. farinae belonged to 26 phyla, 43 classes, 100 orders, 167 families and 284 genera. The bacteria in D. farinae were mainly annotated to five phyla of Proteobacteria, Firmicutes, Bacteroidota, Actinobacteriota, and Acidobacteriota, with Proteobacteria as the dominant phylum, and mainly annotated to five dominant genera of Ralstonia, norank-f-Mitochondria, Staphylococcus and Sphingomonas, with Wolbachia identified in the non-dominant genus. CONCLUSIONS: A high diversity is identified in the composition of the bacterial community in D. farinae, and there are differences in bacterial community diversity and abundance among D. farinae.

[Changes in the related indicators of bone formation and bone resorption in severely burned rats].

Objective: To observe the changes in the related indicators of bone formation and bone resorption in severely burned rats. Methods: The experimental research method was adopted. Thirty female Sprague-Dawley rats aged 6 to 8 weeks were divided into sham injury group, 12% total body surface area (TBSA) full-thickness burn group, and 24%TBSA full-thickness burn group according to the random number table, with 10 rats in each group. The rats were treated on the back correspondingly, after which, the burned rats were rehydrated by intraperitoneal injection according to the Parkland formula, and the wound was coated with 20 g/L iodophor until wound healing. On post injury day (PID) 28, the tibia tissue of rats in each group was collected. The new bone tissue and the number of osteoclasts were observed after staining with Masson and tartrate-resistant acid phosphatase, respectively. The abdominal aortic blood of rats in each group was harvested for serum preparation. The bone metabolism indexes of serum calcium ion and phosphorus ion concentration were determined by the methyl thymol blue colorimetric method and phosphomolybdic acid method, respectively. The serum levels of bone formation marker of aminoterminal propeptide of type 1 procollagen (P1NP) and bone resorption marker of beta-carboxy-terminated peptide of type Ⅰ collagen (ß-CTX) were determined by enzyme-linked immunosorbent assay. The first lumbar spine tissue of rats in each group was collected, and the mRNA expression levels of osteoprotegerin, receptor activator of nuclear factor-κB ligand (RANKL), tumor necrosis factor receptor-associated factor 6 (TRAF-6), nuclear factor of activated T cell 1 (NFATC1), c-Fos, and c-Src were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were statistically analyzed with one-way analysis of variance, Bonferroni method, Welch test, Games-Howell test, Kruskal-Wallis test, Mann-Whitney U test, and Bonferroni correction. Results: On PID 28, compared with that in sham injury group, the formation of new bone tissue in the tibia tissue of rats in the two burn groups was decreased, and the larger the burn area, the more obvious the decrease. The numbers of osteoclasts in the tibia tissue of rats in the two burn groups were similar, both significantly more than the number in sham injury group. On PID 28, the serum calcium ion concentration and serum level of ß-CTX of rats in the three groups were similar (P>0.05). The serum phosphorus ion concentration of rats in 24%TBSA full-thickness burn group was significantly higher than that in 12%TBSA full-thickness burn group (P<0.05), and the serum phosphorus ion concentrations in the two burn groups were significantly higher than the concentration in sham injury group (P<0.01). The serum level of P1NP of rats in 24%TBSA full-thickness burn group was significantly lower than that in sham injury group (P<0.01). On PID 28, the mRNA expression levels of osteoprotegerin in the first lumbar spine tissue of rats in sham injury group, 12%TBSA full-thickness burn group, and 24%TBSA full-thickness burn group were 1.01±0.20, 1.71±0.83, and 2.24±0.51, respectively, and that in 24%TBSA full-thickness burn group was significantly higher than that in sham injury group (P<0.01). The mRNA expression level of RANKL in the first lumbar spine tissue of rats in 24%TBSA full-thickness burn group was 1.31±0.17, which was significantly higher than 1.00±0.14 in sham injury group and 0.97±0.10 in 12%TBSA full-thickness burn group (P<0.01). The mRNA expression levels of TRAF-6, NFATC1 (Z=3.141, 3.782), and c-Src in the first lumbar tissue of rats in 12%TBSA full-thickness burn group and 24%TBSA full-thickness burn group and the mRNA expression level of c-Fos in the first lumbar tissue of rats in 12%TBSA full-thickness burn group were significantly higher than those in sham injury group (P<0.05 or P<0.01). The mRNA expression levels of c-Fos and c-Src in the first lumbar spine tissue of rats in 12%TBSA full-thickness burn group were significantly higher than those in 24%TBSA full-thickness burn group (P<0.01). Conclusions: Severe burns can cause a decrease in the generation of new bone tissue, an increase in the number of osteoclasts and the serum phosphorus ion concentration, and a decrease in the serum level of P1NP in rats. The level of osteoprotegerin, RANKL, TRAF-6, NFATC1, c-Fos, and c-Src in bone tissue showed an increasing trend while the level of NFATC1, c-Fos, and c-Src showed a decreasing trend with the increase of burn area.

[Effects and mechanism of estrogen receptor ß agonist on the migration and oxidative stress of human umbilical vein endothelial cell under high glucose condition].

Objective: To investigate the effects and related mechanism of estrogen receptor ß (ERß) agonist on the migration and oxidative stress of human umbilial vein endothelial cells (HUVECs) under high glucose condition. Methods: The experimental research method was adopted. HUVECs were routinely cultured and passaged, and then cells of the logarithmic growth phase were collected for the subsequent experiments. The cells were divided into three groups according to the random number table, including normal control group (cultured with Roswell Park Memorial Institute 1640 cell culture medium (the same cell culture medium below) containing 5.5 mmol/L D-glucose), high glucose alone group (cultured with cell culture medium containing 25.0 mmol/L D-glucose alone), and high glucose+ERß agonist diarylpropionitrile (DPN) group (cultured with cell culture medium containing 25.0 mmol/L D-glucose and 10 µmol/L DPN). Scratch test was conducted to detect the cell migration rate in the 3 groups at 24 h post scratching. Fluorescent probe method was used to detect the reactive oxygen species (ROS, denoted by red fluorescence intensity) of cells in the 3 groups on 5 d post culture. Western blotting was used to detect the protein expression levels of vascular endothelial growth factor (VEGF) and superoxide dismutase 2 (SOD2) of cells in the 3 groups on 5 d post culture. In the above-mentioned experiments, cells were grouped and cultured correspondingly as before, the number of samples in each group was 5. Data were statistically analyzed with one-way analysis of variance and least significant difference t test. Results: At 24 h post scratching, the cell migration rate in high glucose alone group was (36±5)%, which was significantly lower than (76±4)% of normal control group and (65±5)% of high glucose+DPN group (t=14.511, 9.603, P<0.01), and the cell migration rate in high glucose+DPN group was significantly lower than that in normal control group (t=3.943, P<0.01). On 5 d post culture, the level of ROS of cells in high glucose alone group (1.81±0.12) was significantly increased compared with normal control group and high glucose+DPN group (1.00±0.14, 0.91±0.15, t=9.679, 10.549, P<0.01), while the level of ROS of cells in normal control group and high glucose+DPN group were close (t=1.031, P>0.05). On 5 d post culture, the protein expression levels of VEGF and SOD2 of cells in high glucose alone group were significantly lower than the levels of normal control group (t=14.175, 13.787, P<0.01) and high glucose+DPN group (t=6.321, 17.750, P<0.01). The protein expression level of VEGF of cells in high glucose+DPN group was significantly lower than the level of normal control group (t=7.206, P<0.05), while the protein expression level of SOD2 of cells in high glucose+DPN group was significantly higher than the level of normal control group (t=2.890, P<0.05). Conclusions: The activation of ERß can improve the inhibition of HUVECs migration induced by high glucose and alleviate oxidative stress injury induced by high glucose, which may be achieved by promoting the expression of VEGF and SOD2.

[Research advances on the effect of hydrogen peroxide in wound healing].

Wound healing is a highly ordered biological process involving a variety of cells, fluids, and molecules. Any obstacles in the link may cause poor wound healing and even the formation of chronic wounds. In recent years, more and more studies have reported that hydrogen peroxide plays an important role in wound healing and throughout the whole process of wound healing. This article reviews the research progress of hydrogen peroxide and literatures in the related fields to provide new ideas and clues for promoting the basic and clinical research of wound healing.

Elevated hepatic MDR3/ABCB4 is directly mediated by MiR-378a-5p in human obstructive cholestasis.

OBJECTIVE: The function of MDR3 is important in bile acid transport. The miRNAs can suppress the expression of gene through combining mRNA of target gene. The regulation about MDR3 mediated by FXR or PPARα in cholestasis is clear, but the mechanism through miRNA is hardly reported. We aimed to find out the miRNA, which could suppress MDR3 expression and the significance of this connection in cholestasis. PATIENTS AND METHODS: We measured hsa-miR-378a-5p expression level in liver tissues from 20 patients with cholestasis and 15 patients without cholestasis by quantitative PCR. We also tested the level of clinical features of the same group. HepG2 cell lines were performed experiments to discover the connection between hsa-miR-378a-5p and MDR3, including transient transfection, RNA and protein extraction, qPCR, Western blotting and luciferase reporter assay. RESULTS: A significant decrease of miR-378a-5p was observed in obstructive cholestasis patient liver tissues compared to control group. We also find that the miR-378a-5p expression is correlated to several clinical features, which are important biomarkers in cholestatic liver injury. Then we predicted that MDR3 may be the target gene of miR-378a-5p through miRanda v3.3a. We programed the transient transfection of mimics and inhibitor on HepG2 cell lines, and detected the mRNA and protein expression of MRP2, MRP3 and MDR3. The results suggested that miR-378a-5p could negatively regulate MDR3 expression in both mRNA and protein expression level, and this regulation is specific. We didn't find same regulation in MRP2 and MRP3. Dual luciferase assays proved this regulation is mediated by a direct binding between miR-378a-5p and CDS of MDR3. CONCLUSIONS: We found that hsa-miR-378a-5p expression was down-regulated in cholestatic liver tissues, compared to control liver tissues. Transient transfection and luciferase reporter assay in HepG2 cell lines results suggest that hsa-miR-378a-5p can directly combine MDR3 mRNA and suppress MDR3 protein expression. The down-regulated hsa-miR-378a-5p may cause a protective alteration through up-regulating MDR3 expression in cholestasis.

TTTCA repeat expansion causes familial cortical myoclonic tremor with epilepsy.

BACKGROUND AND PURPOSE: The aim was to investigate whether abnormal TTTTA and TTTCA repeat expansions in introns of SAMD12, TNRC6A and RAPGEF2 are involved in the pathogenesis of familial cortical myoclonic tremor with epilepsy (FCMTE). METHODS: Five families diagnosed with FCMTE were included in the current genetic analysis. Whole-exome sequencing was performed in selected patients of each family. TTTTA and TTTCA expansions were examined by repeat-primed polymerase chain reaction. The clinical features of FCMTE were elicited as defined by the common genetic mechanism of 14 patients. RESULTS: Abnormal TTTCA expansion was identified and co-segregated in all five FCMTE families, four inserted in SAMD12 and one in RAPGEF2. The insertion of expanded TTTCA was not found in 116 control alleles. TTTTA expansion in SAMD12 was detected in 90.9% (10/11) of patients or mutation carriers; TTTTA expansion in RAPGEF2 was not found. The onset age of myoclonic tremor was 27.4 ± 5.9 (19-37) and epilepsy usually presented around age 34. Focal and generalized seizures were witnessed with various origins recorded by electroencephalogram. Cognitive deficits were not common within the first 3 years after epilepsy onset. Emotional instability was reported by most patients. No patients showed any cerebellar deficits. Valproate added with clonazepam is effective in controlling seizures but cannot guarantee a complete remission of tremor. Repeat length showed intergenerational instability and was inversely correlated with age at onset of myoclonic tremor and epilepsy. CONCLUSIONS: TTTCA expansion insertion is associated with FCMTE in Chinese families. The homogenous genetic mechanism allowed for a higher precision of FCMTE description.

Mitochondrial protein OPA mediates osteoporosis induced by radiation through the P38 signaling pathway.

OBJECTIVE: Bone marrow is full of mitochondria. However, the role of bone marrow mitochondrial protein in bone marrow damage and related signal transduction mechanism remains to be further studied. OPA is a newly discovered mitochondrial transmembrane protein. Its expression pattern and function in the physiological and pathological conditions of bone marrow are still elusive. The purpose of this study is to investigate the potential role of OPA in osteoporosis. PATIENTS AND METHODS: A mouse osteoporosis model was established by radiation. The OPA expression was tested by Western blot and qRT-PCR. The P38 signaling activity was evaluated by enzymatic activity kit. The mitochondrial ATP production was determined by flow cytometry. The bone marrow cell apoptosis was detected by flow cytometry. U0126 was used to pretreat mouse before modeling. Bone marrow tissue was collected from patients who received osteoporosis surgery to test the OPA expression, P38 activation and cell apoptosis. The OPA and P38 levels were analyzed by correlation. RESULTS: The mouse osteoporosis model was successfully established by radiation induction. In this osteoporosis model, the expression of OPA was increased. The P38 signaling was activated while the mitochondrial ATP production was reduced, with the increase of apoptosis of bone marrow cells. By contrast, U0126 pretreatment markedly inhibited the OPA expression, restrained the P38 signaling pathway, enhanced mitochondrial ATP production and suppressed the bone marrow cell apoptosis in mouse osteoporosis model. A significantly positive correlation was found between OPA and P38. CONCLUSIONS: The down-regulation of OPA inhibits cell apoptosis and improves osteoporosis via inducing mitochondrial ATP production and suppressing the P38 signaling pathway.

[Research progress of hydrogen-rich saline for eye diseases].

As an antioxidant, hydrogen-rich water has been widely studied in recent years. It is mild enough neither to disturb metabolic redox reactions nor to affect signaling reactive oxygen species. Therefore, there should be no or few adverse effects of H(2). Numerous studies in biology and medicine show that hydrogen-rich water has achieved good results in nervous system, digestive system, respiratory system, heart and kidney diseases. At present, there are few studies on the treatment of eye diseases with hydrogen-rich water. This review summarizes the research progress of hydrogen-rich water for eye diseases in the domestic and foreign literatures. (Chin J Ophthalmol, 2018, 54: 631-635).

[Clinical analysis of 138 multiple primary cancers diagnosed of digestive system malignant tumor initially].

Objective: To study the clinical characteristics, strategy of treatment and prognosis of multiple primary cancers(MPC) diagnosed of digestive system malignant tumor firstly. Methods: From January, 2000 to December, 2015, the clinical, follow-up and prognostic data of 138 MPC patients diagnosed of digestive system malignant tumor firstly were retrospectively analyzed. Results: 138 cases were found in 10 580 cases with malignant tumors, and the incidence was 1.30%. There were 129 cases of duplex primary cancers, 8 cases of triple primary cancers and 1 case of quintuple primary cancers. The repetitive primary cancer was occurred in digestive system (61cases, 44.2%) most frequently, with the next in respiratory system (46 cases, 33.3%). 52.2% (72 cases) suffered second primary cancer in 2 years after first primary cancer diagnosed, and 75.4% (104 cases) in 5 years. The median overall survival in patients with all cancer lesions radically treated was 168 months, better than any other treatment (68 months, P<0.05). Conclusions: The second primary cancers of MPC cases initially diagnosed of digestive system malignant tumor most frequently occurred in the digestive system and respiratory system. More concern should be attracted in follow-up, especially in the first 5 years. The key to improve patient' prognosis was radical treatment to every primary cancer.

Combination of histidine, lysine, methionine, and leucine promotes ß-casein synthesis via the mechanistic target of rapamycin signaling pathway in bovine mammary epithelial cells.

The ratio of different AA in the diets of cows is vital to improve milk protein yield. ß-Casein is one of the important milk proteins with high nutritive value. However, the suitable ratio of essential amino acids (EAA) for the expression of ß-casein in the immortalized bovine mammary epithelial cell line is not fully characterized. This study employed response surface methodology to determine the optimal ratio of His, Lys, Met, and Leu on ß-casein expression level in vitro and clarified the effect of the 4 EAA on ß-casein via the mechanistic target of rapamycin (mTOR) signaling pathway. A central composite design containing 5 axial points per EAA and 28 combinations of the 4 EAA was used in our study. The results of response surface methodology and the changes of the mTOR-related signaling proteins were determined by western blot. The results showed that ß-casein level was significantly affected by all 4 EAA (R2 = 0.71). The optimum conditions for ß-casein expression are found to be 5.47 mM of His, 7.48 mM of Lys, 1.17 mM of Met, and 8.21 mM of Leu (His:Lys:Met:Leu = 5:6:1:7) in the designed scope of concentration. The interaction of Leu and Met significantly affected ß-casein expression (P < 0.01). The phosphorylation of mTOR (Ser2481), regulatory associated protein of target of rapamycin (Ser792), ribosomal protein S6 kinase 1 (Thr389), ribosomal protein S6 (Ser235/236), and eukaryotic elongation factor 2 (Thr56) was increased with the supplementation of either single EAA or an optimal combination of EAA. However, the phosphorylation of eukaryotic initiation factor 4E binding protein 1 (Thr37) was decreased with the addition of Lys, Met, or Leu alone. Furthermore, the phosphorylation (P) of eIF2α (Ser51) was decreased when Met was supplemented alone. Under the optimal mixture of 4 EAA, the phosphorylation of mechanistic target of rapamycin complex 1 signaling proteins was significantly greater than the single EAA supplementations and the expression of ß-casein was 98% as high as the positive control (i.e., medium with all AA). A similar trend was found with P-ribosomal protein S6 kinase 1 and P-ribosomal protein S6. In conclusion, the extracellular concentrations of His, Lys, Met, and Leu at a ratio of 5:6:1:7 maximized ß-casein expression in the immortalized bovine mammary epithelial cell line may occur via activation of the mechanistic target of rapamycin complex 1 signaling pathway.

[Association between single nucleotide polymorphism of adenylyl cyclase 3 and essential hypertension].

OBJECTIVE: To explore the association between the tag single nucleotide polymorphism (tag SNP) of the adenylyl cyclase 3 (ADCY3) and the essential hypertension (EH). METHODS: From April to July 2013, a total of 1 061 subjects diagnosed with EH and 1 218 control subjects were recruited from Ningbo, Zhejiang Province. Information was collected by face-to-face interview. Twelve tag SNPs were detected by ligase detection reaction technique. RESULTS: After adjusted for age, gender, body mass index and other related factors, logistic regression analysis showed that 3 loci (rs11689546, rs7593130, rs2241759)were associated with EH. AG genotype of rs11689546 was associated with 0.494 times lower risk of EH (OR=0.494, 95%CI 0.246-0.993; compared with AA genotype). CT genotype of rs7593130 was associated with 1.596 times higher risk of EH (OR=1.596, 95%CI 1.009-2.524; compared with TT genotype), and CT/CC genotype of rs7593130 was associated with 1.627 times higher risk of EH (OR=1.627, 95%CI 1.034-2.559; compared with TT genotype). AG genotype of rs2241759 was associated with 0.669 times lower risk of EH (OR=0.669, 95%CI 0.503-0.891; compared with AA genotype), and CT/CC genotype of rs2241759 was associated with 0.687 times lower risk of EH (OR=0.687, 95%CI 0.518-0.911; compared with TT genotype). CONCLUSION: The polymorphisms of ADCY3 are associated with lower (G allele of the rs11689546 locus and G allele of the rs2241759 locus) or higher (C allele of the rs7593130 locus) risk of essential hypertension.

Construction of recombinant adenovirus Ad-rat PLCg2-shRNA and successful suppression of PLCg2 expression in BRL-3A cells.

Phospholipase Cg2 (PLCg2) induces apoptosis of immune and tumor cells; however, it remains unclear whether PLCg2 promotes hepatocyte apoptosis during liver regeneration (LR). Therefore, to establish a framework for further exploring the function of PLCg2, we generated recombinant adenoviruses carrying a template encoding short hairpin (sh)-RNA targeting PLCg2 (Ad-PLCg2-shRNA), which were used to silence the expression of PLCg2 in BRL-3A cells. First, three pairs of PLCg2-shRNAs were designed, synthesized, and cloned into a shuttle vector, pHBAd-U6-GFP, after annealing. The recombinant shuttle plasmids were co-transfected with the backbone vector pHBAd-BHG into HK293 cells to package the recombinant Ad-PLCg2-shRNAs used to infect BRL-3A cells. Infection efficiency was monitored by observing the number of GFP-positive cells under a fluorescent microscope. To determine the recombinant adenoviruses with the highest silencing efficiency, levels of PLCg2 mRNA were evaluated by qRT-PCR. DNA sequencing confirmed that the correct shRNA coding sequences were inserted into the shuttle vectors and adenoviral plasmids. The titers of three recombinant adenoviruses were at least 1 x 10(10) PFU/mL. The most effective adenoviral construct, with interference efficiency of 77%, was determined by qRT-PCR. These results show that a recombinant adenovirus, Ad-PLCg2-shRNA, was developed and was effective at silencing the rat PLCg2 gene. This construct may contribute to the study of PLCg2 in hepatocyte apoptosis during LR.