[Modified gasless trans-subclavian approach endoscopic lateral neck dissection for treatment of papillary thyroid carcinoma: a series of 31 cases].

Objective: To examine the feasibility of the modified gasless trans-subclavian approach endoscopic thyroidectomy for lateral neck dissection (LND) in papillary thyroid carcinoma (PTC). Methods: The clinical data of 31 patients with PTC who underwent modified gasless trans-subclavian approach endoscopic LND in the Department of Head and Neck Surgery, Run Run Shaw Hospital, from January to October 2022 were retrospectively analyzed. There were 2 males and 29 females, aged (32.6±8.3) years (range: 17 to 55 years). The maximum diameter of the primary thyroid lesion (M(IQR)) was 1.06 (1.16) cm (range: 0.53 to 2.44 cm), and the maximum diameter of the metastatic lymph node was (1.04±0.37) cm (range: 0.44 to 1.88 cm). Operation time, postoperative hospital stay, number of lymph nodes dissected, and postoperative complications were recorded. Outpatient follow-up was conducted until November 30, 2022. Results: All operations were successfully completed with the endoscopy approach without conversion to open surgery. The operation time was 160 (20) minutes (range: 100 to 215 minutes), and the postoperative hospital stay was 4 (2) days (range: 2 to 14 days). The number of lymph nodes obtained by dissection in the central and lateral compartment of the neck was 11 (12) (range: 0 to 37) and 34.7±14.8 (range: 15 to 69), respectively. Temporary hypoparathyroidism occurred in 4 cases and all recovered within 1 month after the operation. One case suffered from recurrent laryngeal nerve injury (continuing followed up to assess whether it is a temporary injury). The complication of LND included 1 case of chylous leakage that was recovered with conservative treatment, 1 case of Horner syndrome returned to normal 3 months after surgery. During follow-up, there was no residual tumor or recurrence. Conclusion: The modified gasless trans-subclavian approach endoscopic LND for PTC is feasible, with a thorough dissection and concealed incision.

[The value of renal color Doppler ultrasound in evaluating the severity and prognosis of patients with acute organophosphorus pesticide poisoning complicated by acute kidney injury].

Objective: To analyze the value of renal color Doppler ultrasound examination and clinical indicators in evaluating the severity and prognosis of acute organophosphorus pesticide poisoning (AOPP) complicated by acute kidney injury (AKI) . Methods: In November 2019, 86 AOPP patients complicated by AKI who were admitted from May 2018 to May 2019 were selected as the observation group, and they were divided into AKI stage 1 group (n=37) , AKI stage 2 group (n=32) and AKI stage 3 group (n=17) . 40 healthy people were selected as the control group. The differences in power Doppler ultrasound (PDU) score, renal interlobular artery resistance index (RI) value and related clinical indicators of each group were measured and analyzed, and the correlations between the indicators were analyzed. At the same time, binary logistic regression was used to analyze the risk factors of death in AOPP patients complicated by AKI. Results: There were statistically significant differences in Acute Physiology and Chronic Health Evaluation (APACHE) Ⅱscore, mean arterial pressure (MAP) , serum creatinine (SCr) and the length of continuous renal replacement therapy (CRRT) between different groups (P<0.05) . Compared with the control group, the APACHE Ⅱscores and SCr of patients in the AKI stage 2 and resistance index AKI stage 3 groups increased, while the MAP decreased (P<0.05) . Compared with the control group, AKI stage 1 group and AKI stage 2 group, the PDU score of patients in the AKI stage 3 group was significantly decreased, and the renal interlobular artery RI value was significantly increased (P<0.05) . SCr was positively correlated with the RI value of renal interlobular arteries and CRRT days (r=0.435, 0.713, P<0.05) , and was negatively correlated with renal PDU score (r=-0.643, P<0.05) . The renal PDU score was negatively correlated with the RI value of renal interlobular arteries and CRRT days (r=-0.350, -0.556, P<0.01) . Binary logistic regression analysis showed that SCr (OR=1.017, 95%CI: 1.004-1.041) and APACHE Ⅱ score (OR=1.289, 95%CI: 1.019-1.827) were risk factors for death in patients with AOPP complicated by AKI (P<0.05) . Conclusion: Both PDU score and the RI value of renal interlobular artery can reflect the severity and stage of patients with AOPP complicated by AKI to a certain extent, but neither of them is a key factor affecting the death of patients.

Endometrial carcinoma: diffusion-weighted imaging diagnostic accuracy and correlation with Ki-67 expression.

AIM: To evaluate the role of diffusion-weighted imaging (DWI) in the detection of endometrial carcinoma and to correlate the apparent diffusion coefficient (ADC) value with Ki-67 expression. MATERIALS AND METHODS: Fifty-two patients with invasive cancer who underwent pelvic MRI were prospectively evaluated using DWI with b-values of 0 and 1000 s/mm2.The ADC values from standard DWI were measured. The expression of Ki-67 in histological specimens was analysed using immunohistochemistry. The ADC values of endometrial carcinoma and normal endometrial parenchyma were compared. Relationships between ADC values and Ki-67 expression were determined using Wilcoxon's signed rank test and the Kruskal-Wallis test. RESULTS: Endometrial carcinoma was detected at DWI as a hyperintense area in 92.3% (48/52) of patients. There was a significant difference in the mean ADC values between endometrial carcinoma and normal endometrial parenchyma (1.39±0.27×10-3 versus 0.93±0.21×10-3 mm2/s, p<0.001). The mean ADC values of grade 1 patients were significantly higher than those of grade 3 patients (1.01±0.16×10-3 versus 0.83±0.21×10-3 mm2/s, p<0.05). The mean ADC values of stage IB patients were significantly lower than those of stage IA patients (0.86±0.16×10-3 versus 1.04±0.21×10-3 mm2/s, p<0.01). The mean ADC values of high Ki-67 expression patients were significantly lower than those of low Ki-67 expression patients (0.82±0.12×10-3 versus 1.16±0.12×10-3 mm2/s, p<0.001). There was a significant negative correlation between the mean ADC value and Ki-67 expression (r=-0.82, p<0.001). CONCLUSION: The ADC value was a helpful parameter for detecting the tumour grade, stage, and proliferation of endometrial carcinoma, and may further improve patient prognosis and contribute to the development of more effective treatment programmes.

[Effect of picroside â ¡ on the expression of mitochondrial VDAC1 after cerebral ischemia/reperfusion in rats].

Objective: To explore the effect of picroside Ⅱ on the expression of mitochondrial voltage-dependent anion channel 1 (VDAC1) in rats after cerebral ischemiareperfusion. Methods: A total of 70 Wistar rats models with middle cerebral artery occlusionreperfusion (MCAO/R) were randomly divided into the sham group, model group, picroside (Picr) group, ruthenium red (RuR) group, RuR+ Picr group, Spermine (Sper) group, Sper+ Picr group (n=10 per group). Modified neurological severity scale (mNSS) was used to evaluated the neurobehavioral function, the expression of reactive oxygen species (ROS) in brain tissues were measured by enzyme-linked immunosorbent assay (ELISA), the morphology of brain tissues was observed by hematoxylin-eosin (HE) staining, the apoptotic cells were counted by terminal deoxynucleotidyl transferase dUTP nick end labeling assay (TUNEL), and the expressions of VDAC1 and endonuclease G (EndoG) were determined by immunohistochemical assay and Western blot. Results: Compared with the shame group, the mNSS scores (9.6±1.9), the expression of ROS[(47.6±2.7)U/ml], the apoptosis of neuron(23.8±2.8), and the expressions of VDAC1(0.94±0.06) and EndoG in cytoplasm (0.76±0.06) and nuclei(0.75±0.06)were enhanced in the model group (all P<0.05). The Picr group had obviously decreased mNSS scores (5.7±0.9), ROS expression[(35.6±2.2)U/ml], number of apoptotic cells (14.5±2.1), VDAC1 (0.63±0.06) and EndoG in cytoplasm (0.34±0.05) and nuclei (0.31±0.06)expressions compared to the model group (P<0.05). Conclusion: Picroside Ⅱ could attenuate cerebral I/R injury by down-regulating the expression of VDAC1 and inhibiting the EndoG release from mitochondria into cytoplasm.

[Inflammatory fibroid polyp of the gastrointestinal tract: a clinicopathologic features of 37 cases].

OBJECTIVE: To study the clinicopathologic features, pathogenesis, and differential diagnosis of inflammatory fibroid polyp (IFP) of the gastrointestinal tract. METHODS: The clinical and pathologic findings of 37 IFPs in the gastrointestinal tract were retrospectively analyzed. Immunohistochemical study and KIT, PDGFRA molecular analysis were carried out and the literatures reviewed. RESULTS: There were 9 males and 28 females. The median age was 57 (range 37 to 78) years. Twenty-two were in the antrum, nine in the ileum, three in the cardia, and one each in the gastric angle, corpora ventriculi and duodenum. The lesion ranged in size from 0.5 to 5.5 cm (mean 3 cm). Grossly, the majority appeared as a solitary non-encapsulated, submucosal, polypoid lesion. There was associated mucosal ulceration in three cases. Microscopically, the gastric lesions showed spindle-shaped cells arranged in an onion skin-like pattern around vessels and mucosal glands in a concentric formation. But the lesions in the ileum represented Vanek's tumor subtype devoid of concentric formations, with spindled to epithelioid cells dispersed in edematous stroma. Most of the lesions were in the mucosa and submucoma, but one small intestinal IFP infiltrated the muscularis propria. The inflammatory component of the lesions consisted predominantly of lymphocytes and eosinophils. Immunohistochemically, all cases displayed diffuse reactivity for vimentin and CD34; and 18 expressed PDGFRA. Analysis of KIT and PDGFRA mutations was performed in 18 cases. No KIT mutations were identified. However, four cases harbored activating mutations in PDGFRA exon 18 (D842V), five showed mutations in exon 12 (p.566-571delSPDGHEinsR). Follow-up in 30 cases showed no recurrence or metastasis. CONCLUSIONS: IFPs not only exhibit two morphologies, but also show mutations in the PDGFRA gene. IFP is a benign mesenchymal tumor rather than a reactive lesion.

Systematic review and meta-analysis comparing three techniques for pancreatic remnant closure following distal pancreatectomy.

BACKGROUND: Established closure techniques for the pancreatic remnant after distal pancreatectomy include stapler, suture and anastomotic closure. However, controversy remains regarding the ideal technique; therefore, the aim of this study was to compare closure techniques and risk of postoperative pancreatic fistula (POPF). METHODS: A systematic review was carried out according to PRISMA guidelines for studies published before January 2014 that compared at least two closure techniques for the pancreatic remnant in distal pancreatectomy. A random-effects model was constructed using weighted odds ratios (ORs). RESULTS: Thirty-seven eligible studies matched the inclusion criteria and 5252 patients who underwent distal pancreatectomy were included. The primary outcome measure, the POPF rate, ranged 0 from to 70 per cent. Meta-analysis of the 31 studies comparing stapler versus suture closure showed that the stapler technique had a significantly lower rate of POPF, with a combined OR of 0.77 (95 per cent c.i. 0.61 to 0.98; P = 0.031). Anastomotic closure was associated with a significantly lower POPF rate than suture closure (OR 0.55, 0.31 to 0.98; P = 0.042). Combined stapler and suture closure had significantly lower POPF rates than suture closure alone, but no significant difference compared with stapler closure alone. CONCLUSION: The use of stapler closure or anastomotic closure for the pancreatic remnant after distal pancreatectomy significantly reduces POPF rates compared with suture closure. The combination of stapler and suture closure shows superiority over suture closure alone.

Association between interleukin-10 polymorphisms and sepsis: a meta-analysis.

Genetic association studies of the cytokine interleukin-10 (IL-10) and sepsis have provided inconsistent results. This work attempts to further quantitatively assess the association of three widely evaluated polymorphisms of IL-10 (-592C/A, -819C/T, -1082A/G) with sepsis susceptibility through a meta-analysis. A search of Pubmed, Web of Science and EMBASE databases was performed. Overall, the three polymorphisms have no strong association with sepsis risk. Subgroup analysis by ethnicity showed there was association between sepsis susceptibility with -592C/A in Caucasians (A vs. C: OR 0·78, 95% CI 0·62-1·00, P = 0·05; AA + CA vs. CC: OR 0·75, 95% CI 0·56-1·00, P = 0·05), and with -1082A/G in Asians (G vs. A: OR 1·41, 95% CI 1·04-1·91, P = 0·03; GG + AG vs. AA: OR 2·11, 95% CI 1·07-4·16, P = 0·03). This meta-analysis suggests that -592C/A and -1082A/G polymorphisms are associated with sepsis susceptibility in Caucasian, and Asian populations, respectively.

Withaferin A attenuates lipopolysaccharide-induced acute lung injury in neonatal rats.

Withaferin A (WFA) is an active compound from Withania somnifera and has been reported to exhibit a variety of pharmacological activities such as anti—inflammatory, immunomodulatory and anti—tumor properties. In the present study, we investigated the potential protective role of WFA on acute lung injury in neonatal rats induced by lipopolysaccharide (LPS). We found that WFA significantly attenuated the pathological changes of lungs induced by LPS injection. Administration with WFA obviously decreased pulmonary neutrophil infiltration accompanied with decreased MPO concentrations. WFA also reduced the expression of pro—inflammatory cytokines including MIP—2, TNF—α, IL—1β and IL—6. Meanwhile, the expression levels of anti—inflammatory mediators such as TGF—β1 and IL—10 were significantly increased following WFA administration. Moreover, WFA protected LPS—treated rats from oxidative damage via up—regulation of TBARS and H2O2 concentrations and down—regulation of ROS contents. Taken together, the present study demonstrated that WFA administration attenuated LPS—induced lung injury through inhibition of inflammatory responses and oxidative stress.

Relationship between zinc and the growth and development of young children.

The purpose of this study was to evaluate the relationship between zinc and the growth and development of young children. The parents of 8102 young children were surveyed in person by a trained surveyor using structured questionnaires. The hair zinc concentration of the children was determined using an atomic absorption spectrophotometer. The height, weight, sitting height, and head circumference of the children were measured at follow-up visits. There was a positive correlation between hair zinc concentration and adaptive developmental quotient (ADQ; r = 0.3164, P = 0.0272) while no correlation was found between hair zinc concentration and body measurement Z scores or intelligence quotient (IQ). There was a strong positive correlation between hair zinc concentration and weight-for-age Z scores (r = 0.3618, P = 0.0416) and ADQ (r = 0.2761, P = 0.0387) in boys; there was no correlation between hair zinc concentration and body measurement Z scores, IQ, and ADQ in girls. In boys with normal hair zinc levels, ADQ was 9.58 (P = 0.0392), higher than in boys who had zinc-deficient hair. In girls with normal hair zinc levels, ADQ was 2.52 (P = 0.0296), lower than in girls with zinc-deficient hair. In conclusion, there is no significant correlation between hair zinc levels and IQ or Z scores for all body measurements in young children.

Connexin43 hemichannels mediate small molecule exchange between chondrocytes and matrix in biomechanically-stimulated temporomandibular joint cartilage.

OBJECTIVE: Connexin (Cx) 43 hemichannels play a role in mechanotransduction. This study was undertaken in order to determine if Cx43 hemichannels were activated in rat temporomandibular joint (TMJ) chondrocytes under mechanical stimulation. METHODS: Sprague-Dawley rats were stimulated dental-mechanically. Cx43 expression in rat TMJ cartilage was determined with immunohistochemistry and real-time PCR, and Cx43 hemichannel opening was evaluated by the extra- and intracellular levels of prostaglandin E2 (PGE2). Both primary rat chondrocytes and ATDC5 cells were treated with fluid flow shear stress (FFSS) to induce hemichannel opening. The Cx43 expression level was then determined by real-time PCR or Western blotting, and the extent of Cx43 hemichannel opening was evaluated by measuring both PGE2 release and cellular dye uptake. RESULTS: Cx43 expression and intra- and extracellular PGE2 levels were increased in mechanically-stimulated rat TMJ cartilage compared to the unstimulated control. The FFSS treatment increased Cx43 expression and induced Cx43 hemichannel opening in primary rat chondrocytes and ATDC5 cells indicated by enhanced PGE2 release and dye uptake. Furthermore, the Cx43 hemichannel opening could be blocked by the addition of 18ß-glycyrrhetinic acid, a Cx channel inhibitor, Cx43-targeting siRNA, or by withdrawal of FFSS stimulation. The migration of cytosolic Cx43 protein to the plasma membrane in ATDC5 cells was still significant after 8 h post 2-h FFSS treatment, and the Cx43 protein level was still high at 48 h, which returned to control levels at 72 h after treatment. CONCLUSION: Cx43 hemichannels are activated and mediate small molecule exchange between TMJ chondrocytes and matrix under mechanical stimulation.

Mannose-binding lectin polymorphisms and the risk of sepsis: evidence from a meta-analysis.

Several studies have evaluated the association between mannose-binding lectin (MBL) polymorphisms and sepsis. However, the results are inconclusive and conflicting. To better understand the roles of MBL polymorphisms in sepsis, we conducted a comprehensive meta-analysis. All relevant studies were searched from PubMed, EMBASE and Web of Knowledge databases, with the last report up to 7 May 2013. Twenty-nine studies addressing four MBL polymorphisms (-550G/C, -221G/C, structure variant A/O, Gly54Asp) were analysed for susceptibility to sepsis and one study for sepsis-related mortality. Overall, significant associations between structure variant A/O and susceptibility to sepsis were observed for AO + OO vs. AA [odds ratio (OR) 1·27, 95% confidence interval (CI) 1·05-1·52, P = 0·01] and O vs. A (OR 1·19, 95% CI 1·02-1·40, P = 0·03). In subgroup analysis based on age group, increased risk was found in the paediatric group in the dominant model (OR 1·72, 95% CI 1·16-2·56, P = 0·007). Moreover, there was a slight association between the +54A/B polymorphism and susceptibility to sepsis in Caucasians (recessive model: OR 10·64, 95% CI 1·24-91·65, P = 0·03). However, no association was observed for -550G/C and -221G/C polymorphisms both overall and in subgroup analysis. For sepsis-related mortality, only one study suggested AO/OO was associated with in-hospital mortality in pneumococcal sepsis patients after controlling for confounding variables. Our meta-analysis indicated that MBL structure variants might be associated with susceptibility to sepsis but further studies with a large sample size should be conducted to confirm these findings.

Transcriptome network-based method to identify genes associated with unruptured intracranial aneurysms.

Intracranial aneurysm is a balloon or sac-like dilatation of blood vessels inside the brain. Despite their importance, the biological mechanisms of intracranial aneurysms are not totally understood. We used public genome-wide gene expression profile data to identify potential genes that are involved in intracranial aneurysm in order to construct a regulation network. Some of the transcription factors and target genes that we identified in this network had been identified as related to intracranial aneurysm in previous studies. We found additional transcription factors and target genes that are apparently related to intracranial aneurysm with this method. The confirmation of previously identified genes and transcription factors supports the usefulness of this transcriptome network analysis for the identification of candidate genes involved in intracranial aneurysm.

First Report of Fruit Stem Anthracnose of Kiwifruit Caused by Glomerella septospora in China.

Kiwifruit (Actinidia spp.) is a very economically important fruit crop in southern China, namely Fengxin County. In 2011 and 2012, a new anthracnose on the fruit stem of kiwifruit with blight symptoms was observed during the blossom and fruit-set periods. This caused considerable blossom and fruit drop. To isolate the causal pathogen, symptomatic fruit-stem tissue at the junction of diseased and healthy tissue was cut into 30-mm-long segments. The segments were surface-sterilized in 70% ethanol solution for 10 s, followed by 0.1% mercuric chloride solution for 3 min, and a final set of three rinses in sterile distilled water. Segments were plated onto PDA and incubated at 25°C on a 12-h alternating light-dark cycle (1). Colonies emerging from plant tissues were then transferred to new PDA plates to obtain pure cultures. Colonies grew at the rate of 13.75 mm/day, and were initially white, then became dark green. After 25 days incubation, black and subglobose perithecia appeared at the center of the colonies. Through microscopic examination of perithecia at 400× magnification, club-shaped asci, usually containing 8 ascospores, were found. Ascospores were cylindrical to narrowly fusiform, straight or slightly curved, mostly 3-septate, and 54.5 to 107.5 × 5.0 to 7.5 µm in size. Combining cultural and morphological characteristics with reports in the literature (2), the pathogen was preliminarily identified as a Glomerella species. For six isolates, the rDNA internal transcribed spacer (ITS) region was amplified with the primers ITS1 and ITS4, and the sequences were deposited in GenBank (JX885687). A BLASTn search of GenBank showed that the ITS sequence of our Glomeralla isolate (JX885687) had 100% homology with that of an isolate of Glomeralla septospora (GU935911). Twenty representative isolates (G. septospora) obtained from infected fruit stem tissues in the blossom period and the fruit-set period, respectively, were selected for pathogenicity tests on detached young fruit stems of kiwifruit (JinKui). Each isolate was inoculated into three fruit stems, each of which was slightly wounded with a sterile scalpel and inoculated with a 5-mm-diameter colonized potato dextrose agar (PDA) plug. Negative control stems were treated with the sterile PDA plugs. The inoculated stems were incubated in a growth chamber at 25°C and 95% relatively humidity. After 4 days, characteristic symptoms of fruit stem anthracnose were observed on all inoculated stems, whereas the controls remained asymptomatic. Pure cultures of G. septospora were recovered from all inoculated fruit stems, fulfilling Koch's postulates. These results showed G. septospora is the pathogen of fruit stem anthracnose of kiwifruit, and to our knowledge, this is the first report of G. septospora causing fruit stem anthracnose of kiwifruit in Fengxin county, Jiangxi Province of China, as well as in the world. References: (1) J. Y. Lu. Plant Pathogenic Mycology. China Agriculture Press, Beijing, China, 2004. (2) A. Sivanesan and W. H. Hsieh. Mycological Research, 97:1523, 1993.

[Effects and mechanism of hydrogen peroxide pretreatment with low molarity on oxidative stress induced apoptosis of mouse bone marrow mesenchymal stem cells].

Objective: To investigate the effects and mechanism of hydrogen peroxide (HP) pretreatment with low molarity on oxidative stress induced apoptosis of mouse bone marrow mesenchymal stem cells (BMSCs). Methods: The experimental research methods were used. BMSCs were isolated and cultured from two 2-week-old male BALB/c mice by the whole bone marrow culture method. The 3rd-7th passages of cells in logarithmic growth phase were used for the experiments after identification. According to the random number table (the same grouping method below), the cells were divided into 0 µmol/L HP group (without HP, the same below), 25 µmol/L HP group, 50 µmol/L HP group, 100 µmol/L HP group, 150 µmol/L HP group, 200 µmol/L HP group, 250 µmol/L HP group, and 300 µmol/L HP group in which cells were treated by the corresponding final molarity of HP, respectively. The apoptosis rate was detected by flow cytometry (n=4) after 24 hours of culture. The cells were divided into 0 µmol/L HP group, 25 µmol/L HP group, 50 µmol/L HP group, and 100 µmol/L HP group in which cells were treated by the corresponding final molarity of HP, respeclively. After 24 hours of culture, the protein expressions of B-lymphoma-2 (Bcl-2) and Bcl-2-related X protein (Bax) were detected by Western blotting, and the Bcl-2/Bax ratio was calculated (n=3). The cells were divided into 0 µmol/L HP group, 25 µmol/L HP group, 50 µmol/L HP group, 100 µmol/L HP group, 200 µmol/L HP group, and 300 µmol/L HP group in which cells were treated by the corresponding final molarity of HP, respectively. After 24 hours of culture, the protein expressions of glycogen synthase kinase-3ß (GSK-3ß) and phosphorylated GSK-3ß (p-GSK-3ß) were detected by Western blotting (n=3). The cells were divided into 0 µmol/L HP group, 50 µmol/L HP group, and 300 µmol/L HP group in which cells were treated by the corresponding final molarity of HP, respeclively, and HP pretreatment group with 50 µmol/L HP being added in advance for 12 h and then 300 µmol/L HP being added. After 24 hours of culture, the morphology and growth of cells were observed by inverted fluorescence microscopy (non-fluorescent condition) and immunofluorescence method, the apoptosis rate was detected by flow cytometry, the protein expressions of Bcl-2, Bax, cysteine aspartic acid specific protease-3 (caspase-3), caspase-9, cleavage caspase-3, cleavage caspase-9, GSK-3ß, and p-GSK-3ß were detected by Western blotting, and the Bcl-2/Bax ratio was calculated, with all the number of samples being 3. Data were statistically analyzed with one-way analysis of variance and Bonferroni test. Results: After 24 hours of culture, compared with that in 0 µmol/L HP group, the apoptosis rate of cells did not change significantly in 25 µmol/L HP group, 50 µmol/L HP group, or 100 µmol/L HP group (P>0.05) but increased significantly in 150 µmol/L HP group, 200 µmol/L HP group, 250 µmol/L HP group, and 300 µmol/L HP group (P<0.01). After 24 hours of culture, compared with that in 0 µmol/L HP group, the Bcl-2/Bax ratio of cells increased significantly in 25 µmol/L HP group and 50 µmol/L HP group (P<0.05 or P<0.01) but decreased significantly in 100 µmol/L HP group (P<0.05). After 24 hours of culture, compared with those in 0 µmol/L HP group, the protein expression of GSK-3ß in cells showed no significant change in 25 µmol/L HP group and 50 µmol/L HP group (P>0.05), the protein expressions of p-GSK-3ß in cells significantly increased in 25 µmol/L HP group and 50 µmol/L HP group (P<0.01), the protein expressions of GSK-3ß and p-GSK-3ß in cells in 100 µmol/L HP group showed no significant change (P>0.05), the protein expressions of GSK-3ß in cells in 200 µmol/L HP group and 300 µmol/L HP group were significantly increased (P<0.05). but the protein expression of p-GSK-3ß in cells in 200 µmol/L HP group and 300 µmol/L HP group was significantly decreased (P<0.05). After 24 hours of culture, the morphology and growth of cells in 0 µmol/L HP group and 50 µmol/L HP group were similar and normal; in contrast, the cells in 300 µmol/L HP group became smaller and round, with the cell protrusions being shorter or disappeared, the nucleus being cavitated, and the cell abscission being increased significantly; the morphology of most cells in HP pretreatment group was normal, with the shedding of cells being less than that in 300 µmol/L HP group, and the morphology of nucleus being normal. After 24 hours of culture, the protein expression of caspase-9 was similar among the four groups (P>0.05). Compared with that in 0 µmol/L HP group, the apoptosis rate and the protein expressions of cleavage caspase-9, caspase-3, and cleavage caspase-3 of cells in 50 µmol/L HP group showed no significant changes (P>0.05), the Bcl-2/Bax ratio of cells in 50 µmol/L HP group increased significantly (P<0.05), the apoptosis rate and the protein expressions of cleavage caspase-9, caspase-3, and cleavage caspase-3 of cells in 300 µmol/L HP group were significantly increased (P<0.01), while the Bcl-2/Bax ratio of cells in 300 µmol/L HP group was significantly decreased (P<0.05). Compared with those in 300 µmol/L HP group, the apoptosis rate and the protein expressions of cleavage caspase-9, caspase-3, and cleavage caspase-3 of cells were significantly decreased in HP pretreatment group (P<0.05 or P<0.01), while the Bcl-2/Bax ratio of cells was significantly increased in HP pretreatment group (P<0.01). After 24 hours of culture, the protein expressions of GSK-3ß and p-GSK-3ß of cells in 0 µmol/L HP group, 50 µmol/L HP group, 300 µmol/L HP group, and HP pretreatment group were 1.09±0.14, 0.62±0.17, 1.35±0.21, 0.74±0.34, 0.68±0.03, 0.85±0.08, 0.38±0.10, and 0.54±0.09, respectively. Compared with those in 0 µmol/L HP group, the protein expression of p-GSK-3ß of cells was significantly increased in 50 µmol/L HP group (P<0.05) but significantly decreased in 300 µmol/L HP group (P<0.01), while the protein expression of GSK-3ß of cells was significantly increased in 300 µmol/L HP group (P<0.05). Compared with those in 300 µmol/L HP group, the protein expression of GSK-3ß of cells was significantly decreased in HP pretreatment group (P<0.01), while the protein expression of p-GSK-3ß of cells was significantly increased in HP pretreatment group (P<0.01). Conclusions: The molarity of 50 µmol/L may be the optimal molarity of HP to pretreat mouse BMSCs, and 50 µmol/L HP pretreatment can antagonize mitochondrial pathway of oxidative stress induced apoptosis by inhibiting the activity of GSK-3ß.

Polymorphisms in the toll-like receptor 9 gene associated with sepsis and multiple organ dysfunction after major blunt trauma.

BACKGROUND: Toll-like receptor (TLR) 9 is the pattern recognition receptor for microbial DNA. Genetic variation within pattern recognition receptors for bacterial endotoxin and exotoxin has been shown to be associated with the risk of sepsis and organ dysfunction in critical illness. However, little is known about the clinical relevance of TLR9 gene polymorphisms in critical illness. METHODS: A total of 557 patients with major blunt trauma were included in the study. Genetic variation data for the entire TLR9 gene were obtained from the HapMap Project. The genotypes of TLR9 gene polymorphisms were determined using a pyrosequencing method. Whole peripheral blood samples obtained immediately after admission were stimulated with bacterial DNA and production of tumour necrosis factor (TNF) α was then determined. Sepsis morbidity rate and multiple organ dysfunction (MOD) scores were assessed. RESULTS: Of five single-nucleotide polymorphisms (SNPs) genotyped, four (rs187084, rs352139, rs352140 and rs352162) existed as common SNPs and were in strong linkage disequilibrium. Both rs187084 and rs352162 were significantly associated with TNF-α production by peripheral blood leucocytes in response to bacterial DNA stimulation and a higher sepsis morbidity rate in patients with major trauma. In addition, the rs352162 polymorphism was significantly associated with MOD scores, whereas rs187084 showed a trend to be associated with MOD score. CONCLUSION: TLR9 polymorphisms rs187084 and rs352162 might be used to provide relevant risk estimates for the development of sepsis and MOD in patients with major trauma.

Cerebral phaeohyphomycosis caused by Bipolaris spicifera after heart transplantation.

Phaeohyphomycosis is an increasingly recognized cause of brain abscess in both immunocompetent and immunocompromised hosts. We report a case of cerebral phaeohyphomycosis in a 55-year-old male heart transplant recipient caused by Bipolaris spicifera. We review the literature regarding the pathogenesis, epidemiology, diagnosis, and management of infections with dematiaceous fungi.

Does endoplasmic reticulum stress stimulate the apoptosis of granulosa cells in polycystic ovary syndrome?

The aim of this study was to determine whether endoplasmic reticulum (ER) stress is involved in the apoptosis of granulosa cells in patients with polycystic ovary syndrome (PCOS). A total of 116 patients undergoing in vitro fertilization/intracytoplasmic sperm injection cycles at the Binzhou Medical University Hospital IVF Center between September 2019 and January 2020 were enrolled in the study. Apoptosis of the granulosa cells in each patient was analyzed using flow cytometry, and progesterone and estrogen levels in the cell-culture fluid were measured by enzyme-linked immunosorbent assay. The expressions of X-box-binding protein 1 (XBP1(s)), activating transcription factor 6 (ATF6), C/EBP homologous protein (CHOP), B-cell CLL/lymphoma 2 protein (Bcl-2), and Bcl-2 associated X protein (Bax) at the gene or protein level were analyzed using quantitative real-time polymerase chain reaction and Western blotting, respectively. In patients with PCOS, body mass index and basal serum concentrations of luteinizing hormone, testosterone (T), and anti-Mullerian hormone significantly increased (P < 0.05). The number of oocytes retrieed and the rate of clinical pregnancy after the first frozen embryo transfer also significantly increased (P < 0.05). Although apoptosis rate in the granulosa cells significantly increased in patients with PCOS, progesterone (P) and estrogen (E2) levels in the cell-culture fluid significantly decreased (P < 0.05). The apoptosis of granulosa cells was also found to affect blastocyst formation. Furthermore, the messenger ribonucleic acid (mRNA) expressions of XBP1(s), ATF6, CHOP, and Bax significantly increased in patients with PCOS, while that of Bcl-2 significantly decreased (P < 0.05). The protein expressions of CHOP and Bax significantly increased in patients with PCOS, while that of Bcl-2 significantly decreased (P < 0.05). After treatment of the granulosa cells with tauroursodeoxycholic acid, apoptosis rate and mRNA or protein expressions of XBP1(s), CHOP, and Bax significantly decreased, while the expression of Bcl-2 and levels of progesterone and estrogen significantly increased (P < 0.05). We conclude that ER stress could induce the apoptosis of granulosa cells in patients with PCOS. Cell apoptosis may decrease the number of blastocysts formed.

[Role of interleukin-6 in human umbilical vein endothelial cell to mesenchymal cell transformation].

Objective: To observe the effect of interleukin-6 (IL-6) on the phenotype and function of human umbilical vein endothelial cells (HUVECs) and explore the role of IL-6 in the process of endothelial-to-mesenchymal transition (EndMT). Methods: The experimental research method was used. Fresh umbilical cord discarded after normal maternal delivery was collected. On the second day of the primary cell isolation and cultivation, the cell morphology was observed under inverted phase contrast microscope. HUVECs of the 4th passage were identified by immunofluorescence method, and 2 batches of HUVECs ofthe 3rd to 5th passages were used for the subsequent experiments. The first batch of cells were divided into 6 groups according to the random number table (the same below): blank control group, 5 ng/mL IL-6 group, 10 ng/mL IL-6 group, 25 ng/mL IL-6 group, 50 ng/mL IL-6 group, and 100 ng/mL IL-6 group. The second batch of cells were divided into 4 groups: blank control group, 10 ng/mL IL-6 group, 25 ng/mL IL-6 group,and 50 ng/mL IL-6 group; the cells in blank control group was cultured with complete culture medium only, while the cells in the other groups were added with IL-6 of the corresponding final mass concentrations.Cells from the 1st batch were cultured for 72 hours after grouping, the morphology of HUVECS in the 6 groups was observed under inverted phase contrast microscope. At 72 h after grouping culture, the positive expressions of coagulation factor Ⅷ and α vascular smooth muscle actin (α-SMA) in HUVECs in the 6 groups were detected by immunofluorescence method, and the ratio of the number of double positive cells to the number of coagulation factor Ⅷ positive cells (the ratio of double positive cells for short) was calculated, with 6 samples per group; mRNA expression levels of vascular endothelial cadherin and α-SMA of HUVECs in 6 groups were detected by reverse transcription-polymerase chain reaction, with 3 samples per group.Cells from the 2nd batch were cultured 72 hours after grouping, the protein expression levels of vascular endothelial cadherin, α-SMA, and type Ⅰ collagen in the 4 groups were detected by Western blotting, with 3 samples per group. Data were statistically analyzed with one-way analysis of variance and Bonferroni correction. Results: On the 2nd day after isolation and cultivation, the primary cells were in short spindle shape or polygon, cells of the 4th passage were identified as HUVECs by immunofluorescence method. At 72 hours of culture after grouping, the cells from the 1st batch in the 6 groups changed to long spindle shape morphologically along with the increase of IL-6 concentration, the intercellular connections decreased or disappeared with the gap between cells becoming larger. At 72 h after grouping culture, compared with that inblank control group, the ratio of double positive cells in 25 ng/mL IL-6 group, 50 ng/mL IL-6 group, and 100 ng/mL IL-6 group were significantly increased (P<0.01); compared with that in 5 ng/mL IL-6 group, the ratio of double positive cells in 25 ng/mL IL-6 group, 50 ng/mL IL-6 group, and 100 ng/mL IL-6 group were significantly increased (P<0.01); compared with that in 10 ng/mL IL-6 group, the ratio of double positive cells in 50 ng/mL IL-6 group and 100 ng/mL IL-6 group were significantly increased (P<0.01); the ratio of double positive cells in 100 ng/mL IL-6 group was significantly increased compared with those in 25 ng/mL IL-6 group and 50 ng/mL IL-6 group (P<0.01). At 72 h after grouping culture, compared with that in blank control group, the mRNA expression levels of vascular endothelial cadherin of cells in 25 ng/mL IL-6 group, 50 ng/mL IL-6 group, and 100 ng/mL IL-6 group were significantly decreased (P<0.01 or P<0.05); compared with that in 5 ng/mL IL-6 group, the mRNA expression levels of vascular endothelial cadherin of cells in 50 ng/mL IL-6 group and 100 ng/mL IL-6 group were significantly decreased (P<0.01); compared with that in 10 ng/mL IL-6 group, the mRNA expression levels of vascular endothelial cadherin of cells in 50 ng/mL IL-6 group and 100 ng/mL IL-6 group were significantly decreased (P<0.01); compared with that in 25 ng/mL IL-6 group, the mRNA expression levels of vascular endothelial cadherin of cells in 50 ng/mL IL-6 group and 100 ng/mL IL-6 group were significantly decreased (P<0.01). At 72 h after grouping culture, compared with that in blank control group, the mRNA expression levels of α-SMA of cells in 5 ng/mL IL-6 group, 10 ng/mL IL-6 group, 25 ng/mL IL-6 group, 50 ng/mL IL-6, group, and 100 ng/mL IL-6 group were significantly increased (P<0.05 or P<0.01). Cells from the 2nd batch were cultured for 72 hours after grouping. Compared with 1.391±0.026 in blank control group, the protein expressions of vascular endothelial cadherin of cells in 10 ng/mL IL-6 group (1.185±0.063), in 25 ng/mL IL-6 group (0.717±0.078), and in 50 ng/mL IL-6 group (0.239±0.064) were significantly decreased (P<0.05); compared with that in 10 ng/mL IL-6 group, the protein expressions of vascular endothelial cadherin of cells in 25 ng/mL IL-6 group and 50 ng/mL IL-6 group were significantly decreased (P<0.01); compared with that in 25 ng/mL IL-6 group, the protein expression of vascular endothelial cadherin of cells in 50 ng/mL IL-6 group was significantly decreased (P<0.01). At 72 h after grouping culture, compared with that in blank control group, the protein expression levels of α-SMA of cells in 10 ng/mL IL-6 group, 25 ng/mL IL-6 group, and 50 ng/mL IL-6 group were significantly increased (P<0.01); compared with that in 10 ng/mL IL-6 group, the protein expression levels of α-SMA of cells in 25 ng/mL IL-6 group and 50 ng/mL IL-6 group were significantly increased (P<0.01). At 72 h after grouping culture, compared with that in blank control group, the protein expressions of type Ⅰ collagen of cells in 25 ng/mL IL-6 group and 50 ng/mL IL-6 group were significantly increased (P<0.05). Conclusions: After IL-6 treatment, the phenotype and function of HUVECS showed the characteristics of mesenchymal cells in a concentration-dependent manner. The inflammatory factor can promote the process of EndMT, and become one of the important factors regulating the mechanism of tissue fibrosis.

NADPH oxidase 4 (Nox4) deletion accelerates liver regeneration in mice.

Liver is a unique organ in displaying a reparative and regenerative response after acute/chronic damage or partial hepatectomy, when all the cell types must proliferate to re-establish the liver mass. The NADPH oxidase NOX4 mediates Transforming Growth Factor-beta (TGF-ß) actions, including apoptosis in hepatocytes and activation of stellate cells to myofibroblasts. Aim of this work was to analyze the impact of NOX4 in liver regeneration by using two mouse models where Nox4 was deleted: 1) general deletion of Nox4 (NOX4-/-) and 2) hepatocyte-specific deletion of Nox4 (NOX4hepKO). Liver regeneration was analyzed after 2/3 partial hepatectomy (PH). Results indicated an earlier recovery of the liver-to-body weight ratio in both NOX4-/- and NOX4hepKO mice and an increased survival, when compared to corresponding WT mice. The regenerative hepatocellular fat accumulation and the parenchyma organization recovered faster in NOX4 deleted livers. Hepatocyte proliferation, analyzed by Ki67 and phospho-Histone3 immunohistochemistry, was accelerated and increased in NOX4 deleted mice, coincident with an earlier and increased Myc expression. Primary hepatocytes isolated from NOX4 deleted mice showed higher proliferative capacity and increased expression of Myc and different cyclins in response to serum. Transcriptomic analysis through RNA-seq revealed significant changes after PH in NOX4-/- mice and support a relevant role for Myc in a node of regulation of proliferation-related genes. Interestingly, RNA-seq also revealed changes in the expression of genes related to activation of the TGF-ß pathway. In fact, levels of active TGF-ß1, phosphorylation of Smads and levels of its target p21 were lower at 24 h in NOX4 deleted mice. Nox4 did not appear to be essential for the termination of liver regeneration in vivo, neither for the in vitro hepatocyte response to TGF-ß1 in terms of growth inhibition, which suggest its potential as therapeutic target to improve liver regeneration, without adverse effects.