[Hinokiol regulates the cell cycle and apoptosis of nasopharyngeal carcinoma CNE1 cells via Hippo-YAP signaling pathway].

Objective: To explore the effects of hinokiol on the cell cyle and apoptosis of CNE1 nasopharyngeal carcinoma cells and the relevant molecular mechanism. Methods: The CNE1 cells were cultured in vitro and incubated with different concentrations of honokiol, and the cells were divided into blank control group, 10 µmol/L, 20 µmol/L and 40 µmol/L hinokiol treatment groups, and 10 µg/ml cisplatin group. Cell viability was determined by methylthiazolyldiphenyl- tetrazolium bromide (MTT) method, the cell cycle distribution was detected by flow cytometry, mitochondrial membrane potential was detected by mitochondrial membrane potential test kit, apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method, and the proteins expression of proliferating cell nuclear antigen (PCNA) and G1/S specific cyclin D1 (cyclin D1) were detected by immunoblotting. RNA-Seq was conducted in the hinokiol-treated cells. The mRNA expression of yes-associated protein delta (YAP) was detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR). The proteins expression of phosphor-YAP (p-YAP) and nuclear YAP were detected by immunoblotting, the nuclear distribution of YAP protein was detected by immunofluorescence in the cells with or without treated with the mammalian STE20-like kinase 1/2 (MST1/2) inhibitor (XMU-MP-1), hinokiol, and XMU-MP-1+hinokiol. Statistical analysis of the data was conducted using GraphPad Prism 8.0 software. Resluts Compared with the control group, the cell viablity of CNE1 cells, the levels of mitochondrial membrane potential, the proteins expression of PCNA and cyclin D1 in hinokiol treatment groups were markedly decreased (all P values<0.05), while the proportion of G0/G1 phase cells and the ratio of TUNEL-positive cells were significantly increased (both P values<0.05). Transcriptome analysis showed that differential genes were mainly enriched in Wnt signaling pathway, tumor necrosis factor pathway, and Hippo signaling pathway. The mRNA level of YAP and the protein expression of YAP in the nucleus were decreased and the level of p-YAP protein was increased in cells treated with hinokiol, which were significantly different from control group (all P values<0.05). Compared with the hinokiol group, XMU-MP-1+hinokiol groups showed the decrease of p-YAP protein expression (1.157±0.076 vs 0.479±0.038, t=37.120, P<0.05), the increase of YAP protein expression in the nucleus (0.143±0.012 vs 0.425±0.031, t=29.181, P<0.05), the reduced proportion of cells in G0/G1 phase [(72.494±3.309)% vs (58.747±2.865)%, t=17.265, P<0.05], and the decrease of apoptosis ratio [(53.158±3.376)% vs (29.621±2.713)%, t=28.584, P<0.05]. Conclusion: Hinokiol can arrest the cell cycle and induce the cell apoptosis of CNE1 cells via Hippo/YAP signaling pathway.

[Neoadjuvant strategy for locally advanced colorectal cancer based organ preservation].

Neoadjuvant therapy for locally advanced colorectal cancer has made great progress in the past 20 years, but there are still limitations such as side effects, organ dysfunction and unsatisfactory control of metastasis. In recent years, with the improvement of surgical techniques and further development of molecular research, how to further improve local control, reduce distant metastasis, and even avoid surgery according to clinical remission to achieve organ preservation, is the current demand and research goal. With the advancement of molecular research, colorectal cancer has different treatment strategies based on microsatellite status. For patients with microsatellite instability locally advanced colorectal cancer, immune checkpoint inhibitor therapy significantly increased the pathologic complete response rate, reduced the incidence of adverse events and improved organ function compared with conventional chemoradiotherapy. For patients with microsatellite stable locally advanced colon cancer, neoadjuvant therapy is still in the exploratory stage. The standard of care is surgery combined with perioperative chemotherapy. For microsatellite stable locally advanced rectal cancer, the complete response rate is improved by enhancing neoadjuvant therapy, which helps to preserve organs. On the other hand, selective radiotherapy preserves organ function and improves quality of life. This article reviews the neoadjuvant treatment strategies for locally advanced colorectal cancer based on organ-sparing strategies.

[A successfully treated case of respiratory failure caused by eating Nassariidaes].

Food poisoning caused by Nassariidaes has occurred frequently in coastal areas of China, especially in summer and autumn. Nassariidaes poisoning can be manifested as lip and tongue paralysis, dizziness, headache, nausea and vomiting, arrhythmia and even respiratory failure. We admitted a case of respiratory failure caused by eating Nassariidaes. After timely respiratory support, hemoperfusion and other active treatment, the patient was recovered and was discharged. This paper summarized clinical characteristics and treatment of Nassariidaes poisoning, in order to provide reference for clinical diagnosis and treatment of similar cases.

Hydroponic barley supplementation fed with high protein diets improves the production performance of lactating dairy cows.

The study investigated the effects of dietary protein level and the inclusion of hydroponic barley sprouts (HB) on lactation performance, blood biochemistry and N use efficiency in mid-lactation dairy cows. Treatments were arranged in a 2 × 2 factorial design with 2 crude protein (CP) levels [16.8% and 15.5% of dry matter (DM)], with HB (4.8% of DM, replacing 4.3% of alfalfa hay and 0.5% of distillers dried grains with solubles (DDGS)) or without HB. Forty-eight multiparous Holstein dairy cows (146 ± 15 d in milk, 40 ± 5 kg/d of milk) were randomly allocated to 1 of 4 diets: high protein diet (16.8% CP, HP), HP with HB (HP+HB), low protein diet (15.5% CP, LP), or LP with HB (LP+HB). An interaction between CP × HB on dry matter intake (DMI) was detected, with DMI being unaffected by HB inclusion in cows fed the high CP diets, but was lower in cows fed HB when the low CP diet was fed. A CP × HB interaction was also observed on milk and milk protein yield, which was higher in cows fed HB with HP, but not LP. Inclusion of HB also tended to reduce milk fat content, and feeding HP resulted in a higher milk protein and milk urea N content, but lower milk lactose content. Feed efficiency was increased by feeding HP or HB diets, whereas N efficiency was higher for cows fed LP or HB diets. There was an interaction on the apparent total-tract digestibility of DM and CP, which was higher when HB was fed along with HP, but reduced when fed with LP, whereas the digestibility of ADF was increased by feeding low protein diets. In conclusion, feeding a low protein diet had no adverse effect on cow performance, while feeding HB improved milk and milk component yield, and N efficiency when fed with a high CP diet, but compromised cow performance with a low CP diet.

Effects of rumen-encapsulated methionine and lysine supplementation and low dietary protein on nitrogen efficiency and lactation performance of dairy cows.

Low crude protein (CP) diets might be fed to dairy cows without affecting productivity if the balance of absorbed AA were improved, which would decrease the environmental effect of dairy farms. The aim of this study was to investigate the effects of supplementing ruminally protected Lys (RPL) and Met (RPM) at 2 levels of dietary CP on nutrient intake, milk production, milk composition, milk N efficiency (MNE), and plasma concentrations of AA in lactating Holstein cows and to evaluate these effects against the predictions of the new NASEM (2021) model. Fifteen multiparous cows were used in a replicated 3 × 3 Latin square design with 21-d periods. The 3 treatments were (1) a high-protein (HP) basal diet containing 16.4% CP (metabolizable protein [MP] balance of -130 g/d; 95% of target values), (2) a medium-protein diet containing 15% CP plus RPL (60 g/cow per day) and RPM (25 g/cow per day; MPLM; MP balance of -314 g/d; 87% of target values), and (3) a low-protein diet containing 13.6% CP plus RPL (60 g/cow per day) and RPM (25 g/cow per day; LPLM; MP balance of -479 g/d; 80% of target values). Dry matter intake was less for cows fed MPLM and LPLM diets compared with those fed the HP diet. Compared with the HP diet, the intake of CP, neutral detergent fiber, acid detergent fiber, and organic matter, but not starch, was lower for cows fed MPLM and LPLM diets. Milk production and composition were not affected by MPLM or LPLM diets relative to the HP diet. Milk urea N concentrations were reduced for the MPLM and LPLM diets compared with the HP diet, indicating that providing a low-protein diet supplemented with rumen-protected AA led to greater N efficiency. There was no significant effect of treatment on plasma AA concentrations except for proline, which significantly increased for the MPLM treatment compared with the other 2 treatments. Overall, the results supported the concept that milk performance might be maintained when feeding lactating dairy cows with low CP diets if the absorbed AA balance is maintained through RPL and RPM feeding. Further investigations are needed to evaluate responses over a longer time period with consideration of all AA rather than on the more aggregated MP and the ratio between Lys and Met.

[Progress in clinical research on potential therapeutic drugs for acute-on-chronic liver failure].

Acute-on-chronic liver failure (ACLF), has a high mortality rate and a poor prognosis. Currently, the only effective treatment for ACLF is liver transplantation. However, the number of patients who can successfully undergo liver transplantation is limited due to the rapid progression of ACLF, the occurrence of serious complications, and a dearth of liver donors. The available drug treatment indication expansion and pathogenesis exploration are expected to delay the progression of ACLF, reduce complications, and provide patients with opportunities for liver transplantation by improving portal vein pressure, inhibiting excessive inflammatory response, correcting energy metabolism disorders, reducing oxidative stress, resisting hepatic cell apoptosis, and promoting liver regeneration.

Odd- and branched-chain fatty acids in milk fat from Holstein dairy cows are influenced by physiological factors.

Dairy products are the major source of odd- and branched-chain fatty acids (OBCFAs), a group of nutrients with emerging health benefits. The animal diet is known to influence milk fat OBCFAs of dairy cows; however, little is known about the effects of physiological factors. The objective of this study was to investigate the effects of parity and lactation stage on OBCFAs in milk fat of dairy cows. Holstein dairy cows (n = 157) were selected according to parity (first, second, third, or greater) and days in milk (DIM) (≤21 DIM, 21 < DIM ≤ 100, 100 < DIM ≤ 200, >200 DIM). All cows were fed the same total mixed ration for three weeks. Milk samples were collected during the last three days of each lactation stage for fatty acid (FA) analyses via gas chromatography. Results showed that first- and second-parity cows displayed significantly higher proportions and yields of iso-14:0, iso-15:0, iso-16:0, total iso-FA, and total branched-chain FA (P < 0.05) compared with other parities. The proportions of C17:0 and C17:1 cis-9 were also greater in first-parity cows (P < 0.05), while the yields of C17:0 and C17:1 cis-9 were similar among different parities (P > 0.05). The proportions of total OBCFAs were greater in first- and second-parity cows (P < 0.05), whereas the highest yield was observed in second-parity cows. Lactation dairy cows in ≤ 21 DIM group displayed lower proportions of iso-13:0, anteiso-13:0, C13:0, iso-14:0, C15:0, iso-16:0, total iso-FA, and total OBCFAs compared with that of the other groups (P < 0.05), and also lower yields of iso-14:0 and iso-16:0 (P < 0.05). In contrast, C17:0 and C17:1 cis-9 proportions and yields were higher in dairy cows with ≤ 21 DIM (P < 0.05). Iso-17:0 and anteiso-17:0 were not affected by lactation stage (P > 0.05). Taken together, our data showed that both parity and lactation stage have considerable effects on milk fat OBCFAs of dairy cows. In summary, first- and second-parity cows had higher milk OBCFAs compared with later parity cows, and OBCFAs with medium chain lengths were lower in dairy cows with ≤ 21 DIM, while C17:0 and C17:1 cis-9 were higher. These findings show that milk OBCFA contents are differentially modulated by physiological state. They will be useful in future studies that seek to alter OBCFA composition of Holstein dairy cow milk fats.

[Effects of CACNA1H gene knockout on autistic-like behaviors and the morphology of hippocampal neurons in mice].

OBJECTIVE: To investigate the effects of CACNA1H gene knockout (KO) on autistic-like behaviors and the morphology of hippocampal neurons in mice. METHODS: In the study, 25 CACNA1H KO mice of 3-4 weeks old and C57BL/6 background were recruited as the experimental group, and 26 wild type (WT) mice of the same age and background were recruited as the control group. Three-chamber test and open field test were used to observe the social interaction, anxiety, and repetitive behaviors in mice. After that, their brain weight and size were measured, and the number of hippocampal neurons were observed by Nissl staining. Furthermore, the CACNA1H heterozygote mice were interbred with Thy1-GFP-O mice to generate CACNA1H-/--Thy1+(KO-GFP) and CACNA1H+/+-Thy1+ (WT-GFP) mice. The density and maturity of dendritic spines of hippocampal neurons were observed. RESULTS: In the sociability test session of the three-chamber test, the KO mice spent more time in the chamber of the stranger mice than in the object one (F1, 14=95.086, P < 0.05; Post-Hoc: P < 0.05), without any significant difference for the explored preference index between the two groups (t=1.044, P>0.05). However, in the social novelty recognition test session, no difference was observed between the time of the KO mice spend in the chamber of new stranger mice and the stranger one (F1, 14=18.062, P < 0.05; Post-Hoc: P>0.05), and the explored preference index of the KO mice was less than that of the control group (t=2.390, P < 0.05). In the open field test, the KO mice spent less time in the center of the open field apparatus than the control group (t=2.503, P < 0.05), but the self-grooming time was significantly increased compared with the control group (t=-2.299, P < 0.05). Morphological results showed that the brain weight/body weight ratio (t=0.356, P>0.05) and brain size (t=-0.660, P>0.05) of the KO mice were not significantly different from those of the control group, but the number of neurons were significantly reduced in hippocampal dentate gyrus compared with the control group (t=2.323, P < 0.05). Moreover, the density of dendritic spine of dentate gyrus neurons in the KO-GFP mice was significantly increased compared with the control group (t=-2.374, P < 0.05), without any significant difference in spine maturity (t=-1.935, P>0.05). CONCLUSION: CACNA1H KO mice represent autistic-like behavior, which may be related to the decrease in the number of neurons and the increase in the density of dendritic spine in the dentate gyrus.

[Survival outcomes and prognostic factors of patients with salvage surgery for hypopharyngeal carcinoma after radiotherapy].

Objective: To investigate the survival outcomes and prognostic factors of patients with salvage surgery for hypopharyngeal carcinoma after radiotherapy. Methods: A retrospective analysis was performed, including 26 patients treated in Ningbo Medical Center Lihuili Hospital between January 2010 and December 2015. All patients were males, aged 48-83 years, of whom 8 cases were local residual after radiotherapy alone, 8 cases were local recurrence after postoperative radiotherapy, 2 cases were residual of cervical lymph nodes after radiotherapy alone, 2 cases were recurrence of cervical lymph nodes after radiotherapy alone, 2 cases were recurrence of cervical lymph nodes after postoperative radiotherapy and 4 cases were recurrence of tracheal stoma. The salvage operations included: local resection, local resection with neck dissection, simple neck dissection, tumor resection of tracheostomy, and additional repair according to the defect. Chi square test was used for recurrence and metastasis analysis, Kaplan-Meier method for survival analysis, Log-rank test for univariate analysis, and Cox regression model for multivariate analysis. Results: The complication rate of salvage surgery was 23.1% (6/26). The recurrence rate was 65.4% (17/26) and the distant metastasis rate was 42.3% (11/26) in the 5-year follow-up after salvage surgery. Patient's age and tumor invasion extent were correlated with recurrence. Initial treatment, tumor persistence or recurrence after radiotherapy, recurrence location and tumor invasion extent were correlated with distant metastasis (all P<0.05). Overall, 3 year and 5 year survival rates were 42.3% and 23.1% respectively. Age, recurrence location, surgical margin and tumor invasion extent were related to prognosis (χ²=6.56, 10.68, 9.32, and 7.90 respectively, all P<0.05). Multivariate analysis showed that surgical margin and tumor invasion extent were independent risk factors for prognosis (OR (95%CI) = 3.19 (1.03-9.84), 14.37 (2.46-84.08), both P<0.05). Conclusion: Salvage surgery is the first choice for patients with recurrence after radiotherapy for hypopharyngeal carcinoma. Safe surgical margin should be ensured, especially in tumors invading muscle, bone tissue or lymph node capsule.

[Predictive value of impedance of leadless pacemaker during implantation on trend changes of pacing threshold].

Objective: To explore the predictive value of the impedance measured during leadless pacemaker Micra implantation on the trend of changes of pacing threshold post implantation. Methods: This is a retrospective cross-sectional study. Patients who received implantation of leadless pacemaker Micra at the Second Xiangya Hospital of Central South University from December 2019 to August 2020 were enrolled. The clinical data and the intraoperative electrical parameters during leadless pacemaker implantation were collected. The impedance and pacing threshold data were analyzed at three time points: immediate release, 5-10 min after release, and after traction test. Receiver operating characteristic (ROC) curves and the area under the curve (AUC) were used to analyze the value of the impedance at immediate release on predicting the trend of changes of pacing threshold post implantation. Results: A total of 21 patients (mean age: (72.2±12.5) years, 12 males) were included. The impedance of 21 patients was (798.1±35.3) Ω immediately after implantation, (800.9±35.6) Ω after 5-10 minutes of release, and (883.6±31.7) Ω after traction test. Impedance was similar between the three time points (P>0.05). The threshold was (0.97±0.11) V/0.24 ms immediately after implantation, (0.95±0.12) V/0.24 ms at 5-10 min after the release, and (0.59±0.06) V/0.24 ms after the traction test. The threshold was significantly lower after the traction test than that immediately after release (P=0.003) and than that at 5-10 minutes after release (P=0.008), suggesting a decreased tendency of the threshold over time. According to the analysis of the ROC curve, the immediate impedance after the release ≥680 Ω could predict the ideal pacing threshold after the traction test (AUC=0.989, 95%CI 0.702-0.964, P<0.001), the prediction sensitivity was 87%, and the specificity was 100%. The pacing threshold would be not ideal with the immediate impedance ≤ 520 Ω (95%CI 0.893-1.000, P<0.001), the sensitivity was 100%, and the specificity was 80%. Conclusions: The impedance immediately after the release has predictive value for the changing trend of threshold post leadless pacemaker Micra implantation. Impedance ≥680 Ω immediately after release is often related with ideal pacing threshold after the traction test. In contrast, the impedance ≤ 520 Ω pacing is often related with unsatisfactory threshold after the traction test, therefore, it is recommended to find a new pacing site to achieve the impedance ≥680 Ω immediately after release during leadless pacemaker Micra implantation.

[Establishment of reference interval of vitamin A for Chinese children and adolescents based on expectation-maximization algorithm].

Objective: To establish the reference interval of vitamin A for Chinese children and adolescents by indirect approach. Method: The serum retinol samples from 28 provinces, autonomous regions and municipalities in China between January 2016 and June 2019 were examined by high performance liquid chromatography in third-party clinical laboratory institutions. Using a cross-sectional study, descriptive statistics of serum vitamin A levels in children and adolescents aged 0-18 years were carried out in population, region and season. The distribution parameters of vitamin A levels were calculated iteratively by the expectation-maximization (EM) algorithm, and the reference interval was established. Results: A total of 1 096 597 cases were included in the analysis, including 631 017 males and 465 580 females, with a median age of 3(1, 6) years. Before the age of 4 years, the median serum vitamin A level increased with age [median level and its quartiles was 0.15 (0.11, 0.18) mg/L for neonates, 0.23 (0.18, 0.29) mg/L for infants aged below 1 year, and 0.27 (0.20, 0.34) mg/L for kids aged 1-<4 years]. The median serum vitamin A level was stable at the age of 4-8 years (median level ranged from 0.27 to 0.29 mg/L). Afterwards, the median serum vitamin A level reached 0.30 mg/L at the age of 9 years, and then an increasing trend with age was observed till 18 years. The median serum vitamin A level was higher in south area (male:0.33 mg/L; female:0.34 mg/L), compared with other areas (0.24-0.30 mg/L). The variation of the median serum vitamin A level among seasons was within 0.03 mg/L. Based on EM algorithm using indirect approach, the reference intervals of vitamin A for Chinese children and adolescents were as follows: newborn, 0.27-0.47 mg/L; infants aged below 1 year, 0.36-0.56 mg/L; children aged 1-<7 years, 0.35-0.54 mg/L; children aged 7-<16 years, 0.46-0.60 mg/L; and teenagers aged over 16 years, 0.38-0.59 mg/L. Conclusion: The EM algorithm can provide a basis for the establishment of vitamin A reference interval for Chinese children and adolescents, and the established reference interval needs to be further verified by direct method.

The effect of androgen on wool follicles and keratin production in Hetian sheep / Efeito do androgênio na produção de folículos e queratina em ovinos Hetianos

Abstract To investigate the optimal androgen concentration for culturing Hetian sheep wool follicle and to detect effects of androgen concentration on wool follicle cell proliferation and apoptosis using immunofluorescence labeling and real-time quantitative fluorescence determinations of wool keratin-associated protein gene expression levels. Wool follicles were isolated by microdissection and wool follicles and skin pieces were cultured in various concentrations of dihydrotestosterone (DHT) in culture medium. Next, daily lengthwise growth measurements of wool follicles were obtained using a microscopic micrometer. Cultured Hetian wool follicles were stained using the SACPIC method to reveal wool follicle structure, while sheep skin slices were used to observe cell proliferation by immunostaining and cell apoptosis using the TUNEL method. At the molecular biological level, keratin-associated protein (Kap) gene expression was studied using wool follicles cultured for various numbers of days in vitro. Effects of androgen concentrations on Hetian wool follicle growth and development were experimentally studied. EdU proliferation assays revealed that androgen promoted cell proliferation within wool follicle dermal papillae. TUNEL apoptosis detection demonstrated that androgen treatment could delay cell apoptosis. Quantitative reverse transcription polymerase chain reaction (qPCR) results demonstrated that gene expression level patterns of Hetian mountain sheep super-high sulfur protein. Kap1.1, KIF1.2, Kap2.12 and Kap4.2 gene expression level of the mountainous experimental group was significantly higher than plains Hetian sheep. An androgen concentration of 100 nM can promote the growth of Hetian wool follicle cells in vitro, resulting in overexpression of some genes of the Kap family.

Resumo Investigar a concentração ideal de andrógenos em cultura de folículos pilosos de carneiro Hetiano e detectar os efeitos da concentração de andrógenos na proliferação e apoptose de células foliculares, por meio de imunofluorescência e de determinação quantitativa, em tempo real, da fluorescência dos níveis de expressão gênica de proteína associada à queratina. Folículos pilosos foram isolados por microdissecção, e folículos de lã e pedaços de pele foram cultivados em várias concentrações de di-hidrotestosterona (DHT) em meio de cultura. Em seguida, medições diárias de crescimento longitudinal dos folículos capilares foram obtidas usando um micrômetro microscópico. Folículos de lã cultivados de Hetianos foram corados pelo método SACPIC para revelar a estrutura do folículo piloso, enquanto fatias de pele de carneiro foram usadas para observar a proliferação celular por imunocoloração e apoptose celular por meio do método TUNEL. Em âmbito da biologia molecular, a expressão gênica da proteína associada à queratina (Kap) foi estudada usando folículos capilares cultivados por vários dias, in vitro. Os efeitos das concentrações de andrógenos no crescimento e desenvolvimento dos folículos de lã de Hetianos foram estudados experimentalmente. Ensaios de proliferação de EdU revelaram que o andrógeno promoveu a proliferação celular dentro das papilas dérmicas do folículo piloso. A detecção de apoptose por TUNEL demonstrou que o tratamento com andrógeno poderia atrasar a apoptose celular. Os resultados da reação em cadeia da polimerase transcrição reversa quantitativa (qPCR) demonstraram que os padrões de expressão gênica da proteína de enxofre Kap1.1, KIF1.2, Kap2.12 e Kap4.2 foram significativamente maiores no grupo de ovinos Hetianos de montanha. Uma concentração de androgênio de 100 nM pode promover o crescimento de células foliculares de lã de Hetianos in vitro, resultando na superexpressão de alguns genes da família Kap.

[Treatment report of three cases of multiple wasp stings].

From September to November 2019, three patients with Hornet stings were admitted to hospital with acute anaphylaxis and various degrees of organ dysfunction. After hemoperfusion combined with continuous hemodialysis and hormone pulse therapy, 1 patient was successfully treated and 2 patients were not cured. Bee Stings can cause systemic allergic reactions and toxic reactions. In severe cases, acute anaphylaxis, rhabdomyolysis, intravascular hemolysis, abnormal coagulation, and acute liver and kidney injury may occur, myocardial damage and other multi-organ function damage or even failure, is a clinical acute and critical illness, mortality is higher. Through the analysis of the diagnosis and treatment process of these 3 cases, the experience is summarized: the Bee sting should be treated as soon as possible, the early prevention and treatment of multiple organ dysfunction, the choice of accurate blood purification and comprehensive treatment, can effectively improve the prognosis, increase the success rate of treatment.

[Effects of stepwise acute pain management on acute pain and post-traumatic stress disorder in children with burns: a prospective randomized controlled study].

Objective: To explore the effects of stepwise acute pain management on acute pain and post-traumatic stress disorder (PTSD) in children with burns. Methods: From November 2018 to December 2019, 196 children with burns who were admitted to West China Hospital of Sichuan University and met the inclusion criteria were enrolled in the prospective randomized controlled study. The children were divided into traditional pain management group (97 children, 51 males and 46 females, aged 1 to 6 years) and stepwise pain management group (96 children, 55 males and 41 females, aged 1 to 6 years) according to the random number table. Children in traditional pain management group were treated with traditional acute pain care, while the children in stepwise pain management group were treated with stepped acute pain management (moderate pain was treated with oral administration of acetaminophen sustained-release dry suspension at the dose of 10 to 15 mg/kg once every 4 to 6 hours, and severe pain was treated with morphine intravenous injection at the dose of 0.1 to 0.2 mg/kg once every 4 hours) on the basis of traditional acute pain care after admission. The COMFORT behavior scale was applied to compare the resting pain levels of children in the two groups within post injury day (PID) 3 (1, 9, and 17 o'clock each day). The adverse reactions of children in the stepwise pain management group during the treatment period were recorded. The occurrence of PTSD within one month after injury was evaluated in both groups by the revised PTSD scale. Data were statistically analyzed with independent sample t test, Bonferroni correction, analysis of variance for repeated measurement, chi-square test, Wilcoxon rank sum test, and Fisher's exact probability test. Results: The pain scores of children in stepwise pain management group were significantly lower than traditional pain management group at 1, 9, and 17 o'clock on PID 1, 1, 9, and 17 o'clock on PID 2, and 1, 9, and 17 o'clock on PID 3 (t=2.71, 3.44, 4.05, 4.18, 4.08, 4.19, 4.25, 3.69, 3.71, P<0.05 or P<0.01). The pain scores of children in both groups showed a decreasing trend over time. Of the 96 children in stepwise pain management group, 84 children were treated with oral administration of acetaminophen sustained-release dry suspension, and 12 children were treated with morphine intravenous injection. No adverse reaction occurred during the treatment period. The incidence of PTSD of children in stepwise pain management group within 1 month after injury was 3.12% (3/96), which was significantly lower than 14.43% (14/97) in traditional pain management group, P<0.05. Conclusions: The stepped acute pain management can relieve the acute pain and reduce the incidence of PTSD in children with burns.

The effect of androgen on wool follicles and keratin production in Hetian sheep.

To investigate the optimal androgen concentration for culturing Hetian sheep wool follicle and to detect effects of androgen concentration on wool follicle cell proliferation and apoptosis using immunofluorescence labeling and real-time quantitative fluorescence determinations of wool keratin-associated protein gene expression levels. Wool follicles were isolated by microdissection and wool follicles and skin pieces were cultured in various concentrations of dihydrotestosterone (DHT) in culture medium. Next, daily lengthwise growth measurements of wool follicles were obtained using a microscopic micrometer. Cultured Hetian wool follicles were stained using the SACPIC method to reveal wool follicle structure, while sheep skin slices were used to observe cell proliferation by immunostaining and cell apoptosis using the TUNEL method. At the molecular biological level, keratin-associated protein (Kap) gene expression was studied using wool follicles cultured for various numbers of days in vitro. Effects of androgen concentrations on Hetian wool follicle growth and development were experimentally studied. EdU proliferation assays revealed that androgen promoted cell proliferation within wool follicle dermal papillae. TUNEL apoptosis detection demonstrated that androgen treatment could delay cell apoptosis. Quantitative reverse transcription polymerase chain reaction (qPCR) results demonstrated that gene expression level patterns of Hetian mountain sheep super-high sulfur protein. Kap1.1, KIF1.2, Kap2.12 and Kap4.2 gene expression level of the mountainous experimental group was significantly higher than plains Hetian sheep. An androgen concentration of 100 nM can promote the growth of Hetian wool follicle cells in vitro, resulting in overexpression of some genes of the Kap family.

[Epidemic characteristics of hand, foot and mouth disease (HFMD) and the lag effect of average daily temperature on the epidemic of HFMD in Beijing].

Objective: To analyze the epidemic characteristics of hand, foot and mouth disease (HFMD) in Beijing and explore the short-term lag effect of daily average temperature on HFMD. Methods: The incidence data and meteorological data of HFMD were collected from Xicheng District, Chaoyang District, and Changping District of Beijing between January 2009 and December 2018. The dose-response relationship and short-term lag effect of temperature and HFMD incidence were explored by the distributed lag non-linear model (DNLM). Results: From 2009 to 2018, a total of 97 210 cases of HFMD were confirmed in Xicheng District, Chaoyang District, and Changping District of Beijing. The incidence rate of HFMD in Xicheng district was 71.83/100 000, which was the lowest in three districts. The total annual average incidence was 146.89/100 000 in the three districts. The cases were concentrated from May to July, with a small peak in October to November. The ratio of male to female was 1.49∶1, and the median age of the patients was 3.08 (1.89, 4.39) years old, which showed a decreasing trend in children under 6 years old (Z=-30.11, P<0.01). The DNLM showed that the cumulative relative risk (RR) curve had a bimodal distribution, with RR of 1.36 (95%CI: 1.05-1.76) and 1.35 (95%CI: 1.10-1.66), and the peak values appeared at 4 ℃ and 26 ℃, respectively. Conclusions: The incidence rate of HFMD in Xicheng was the lowest in three districts of Beijing. In addition, the incidence of HFMD is seasonal, and the incidence is higher in spring and summer than that in autumn and winter. Daily average temperature had a lag effect on HFMD, which was different between high temperature and low temperature.

Elevated CISD2 expression predicts poor diagnosis and promotes invasion and migration of prostate cancer cells.

OBJECTIVE: To explore roles of CDGSH iron-sulfur domain-containing protein 2 (CISD2) in the progression of prostate cancer (PCa) cells, and relationships between CISD2 expression and the prognosis and clinical pathological parameters in PCa patients. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) analysis and Western blot analysis were used to detect the CISD2 expression in PCa tissues and cells. CISD2 siRNA was used to inhibit the CISD2 expression. Kaplan-Meier method and Log rank analysis were performed to determine survival analysis while Chi-square test was performed to analyze the association between CISD2 and clinicopathological parameters of PCa patients. Transwell assay and wound healing assay was conducted to examine the invasion and migration ability of PCa cells, respectively. RESULTS: CISD2 was up-regulated in PCa tissues and cells, and showed positive association with the poor prognosis, T stage, lymphatic invasion, prostate-specific antigen (PSA) level, and distant metastasis of PCa patients. Besides, we found that inhibition of CISD2 significantly impaired the migration and invasion ability of PCa cells. CONCLUSIONS: The paper demonstrated that CISD2 could act as a new target for the diagnosis and treatment of PCa patients.

[Investigation on seroprevalence of Toxoplasma gondii infections among neonates in Fujian Province].

OBJECTIVE: To investigate the seroprevalence of Toxoplasma gondii infections among neonates in Fujian Province, so as to provide insights into the development of interventions for the prevention and control of congenital toxoplasmosis. METHODS: A total of 1 045 neonates delivered in Fujian Province from 2017 to 2018 were recruited, including 387 preterm infants and 658 full-term infants. Umbilical cord blood was sampled from all neonates, and the seroprevalence of anti-T. gondii IgG antibody was detected and compared between preterm and full-term infants. In addition, elbow venous blood samples were collected from neonates'mothers, and the seroprevalence of anti-T. gondii IgG antibody was detected and compared between preterm and full-term infants'mothers. RESULTS: The overall seroprevalence of anti-T. gondii IgG antibody was 9.38% among the 1 045 neonates in Fujian Province. The seroprevalence of anti-T. gondii IgG antibody was 18.35% in the 387 preterm infants, and there was no significant difference in the seroprevalence of anti-T. gondii IgG antibody between male and female infants (17.69% vs. 18.75%, χ2 = 0.07, P > 0.05). The seroprevalence of anti-T. gondii IgG antibody was 4.10% in the 658 full-term infants, and there was no significant difference in the seroprevalence of anti-T. gondii IgG antibody between male and female infants (4.14% vs. 4.08%, χ2 = 0, P > 0.05). In addition, the overall seroprevalence of anti-T. gondii IgG antibody was 15.02% in all neonates' mothers, and the seroprevalence was significantly greater in preterm infants'mothers than in full-term infant's mothers (20.93% vs. 11.55%, χ2 = 16.79, P < 0.01). CONCLUSIONS: The seroprevalence of T. gondii infections is significantly higher in preterm infants and their mothers than in full-term infants and their mothers. Prenatal detection of T. gondii infections and health education pertaining to toxoplasmosis prevention and control knowledge are required to be strengthened to effectively reduce the incidence of congenital toxoplasmosis.

Identification of downstream target genes regulated by CX43 in hepatocellular carcinoma.

The aim of study was to identify the downstream target genes of CX43 by Human Transcriptome Array. Therefore, a gene microarray was generated which consists of CX43-overexpressed hepatocellular carcinoma (HCC) cells transfected with the constructed plasmid and negative controls to identify candidate genes. Integrated bioinformatic analysis was used to clarify biological functions of the identified genes, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, protein-protein interaction network, and survival analysis. The candidate genes were further validated by qRT-PCR in liver cancer tissues and CX43-silenced HCC cells. We have found the mRNA and protein levels of CX43 significantly upregulated in HCC cells transfected with CX43 constructed plasmid. We identified 928 differentially expressed genes including 394 upregulated and 534 downregulated genes, enriched in the cancer related functions and pathways by GO and KEGG pathway analysis. The protein-protein interaction network revealed 9 hub genes in this study. Statistical analysis indicated that upregulation of RALA and SRC was associated with poor prognosis in liver cancer. The differential expression of 2 candidate genes were further validated in HCC cells and tissues. In conclusion, protein-coding genes RALA and SRC could be target genes of CX43 and therapeutic targets for hepatocellular carcinoma.

[A prognostic scoring scale for adult patients with supratentorial primary anaplastic gliomas].

Objective: This study explored the prognostic factors of patients with supratentorial anaplastic gliomas and tried to propose a prognostic scoring scale with aim to provide theoretical reference for clinical treatment. Methods: The clinical data of 198 patients surgically treated for primary anaplastic glioma in Henan Provincial People's Hospital between Jan 2009 and Jan 2018 were reviewed. Univariate and multivariate analyses were used to identify prognostic factors with methods of Kaplan-Meier plot and Cox proportional hazard model, respectively. Based on the prognostic factors, a scoring scale was thereby proposed. Results: Univariate analysis results showed age, tumor location, tumor diameter, preoperative KPS, extent of resection, radiotherapy, chemotherapy, pathology with oligodendroglial components, 1p/19q, IDH, MGMT were significantly correlated with survival (P<0.05). Multivariate analysis results showed age ≥45 years old, tumor diameter ≥6 cm, preoperative KPS<70, without radiotherapy, 1p/19q intact, MGMT promoter unmethylation were independent prognostic risk factors (P<0.05). Patients were scored with 0-6 points based on the formulation that each independent prognostic risk factor was assigned with 1 point. Then patients were further grouped according to the score. Those with less than 2 points were low-risk group, equal to 2 points were medium-risk group, equal to 3 points were high-risk group, more than or equal to 4 points were extremely high-risk group. There were significant differences in survival between the different groups (P<0.000 1). Conclusions: The higher score, the shorter survival time. This prognostic scoring scale can provide a theoretical basis for the prognosis estimation of patients with anaplastic glioma and help to carry out personalized clinical treatment.