Prognostic value of APOBEC3A in patients with cervical squamous cell carcinoma in a major urban center in China: a retrospective study.

Background: APOBEC3A (A3A) has been implicated to have vital prognostic value in several common cancers. This study aimed to investigate the prognostic value of A3A expression in cervical squamous cell carcinoma (CESC). Methods: This retrospective study enrolled 59 patients with CESC or cervical squamous intraepithelial neoplasia from January 2014 to January 2017 in Changhai Hospital, Naval Medical University. Then, A3A histoscores (H-scores) using immunohistochemistry (IHC) were analyzed in formalin-fixed paraffin-embedded archival tissue blocks. Moreover, overall survival was analyzed by the Kaplan-Meier method. Results: The H-score of A3A protein expression was relatively higher in CESC than in squamous intraepithelial neoplasia, and the relative expression level of normal cervical tissues was lower than that of cervical squamous intraepithelial neoplasia (P<0.001). Moreover, the H-score of poorly differentiated cases was 6, which was higher than that of moderately differentiated cases (H-score =3), while the H-score of well-differentiated cases was 2, which was lower than that of moderately differentiated cases. Moreover, patients in the A3A low expression group had higher overall survival rates by prognostic analysis (P=0.027). Conclusions: A3A protein expression was increased during CESC progression. Moreover, A3A expression was tightly related to poor prognosis in CESC. Thus, these results showed that A3A overexpression may provide a marker for poor prognosis in CESC.

Xiaotan Sanjie recipe, a compound Chinese herbal medicine, inhibits gastric cancer metastasis by regulating GnT-V-mediated E-cadherin glycosylation.

OBJECTIVE: Xiaotan Sanjie recipe (XTSJ), a Chinese herbal compound medicine, exerts a significant inhibitory effect on gastric cancer (GC) metastasis. This work investigated the mechanism underlying the XTSJ-mediated inhibition of GC metastasis. METHODS: The effect of XTSJ on GC metastasis and the associated mechanism were investigated in vitro, using GC cell lines, and in vivo, using a GC mouse model, by focusing on the expression of Glc-N-Ac-transferase V (GnT-V; encoded by MGAT5). RESULTS: The migration and invasion ability of GC cells decreased significantly after XTSJ administration, which confirmed the efficacy of XTSJ in treating GC in vitro. XTSJ increased the accumulation of E-cadherin at junctions between GC cells, which was reversed by MGAT5 overexpression. XTSJ administration and MGAT5 knockdown alleviated the structural abnormality of the cell-cell junctions, while MGAT5 overexpression had the opposite effect. MGAT5 knockdown and XTSJ treatment also significantly increased the accumulation of proteins associated with the E-cadherin-mediated adherens junction complex. Furthermore, the expression of MGAT5 was significantly lower in the lungs of BGC-823-MGAT5 + XTSJ mice than in those of BGC-823-MGAT5 + solvent mice, indicating that the ability of gastric tumors to metastasize to the lung was decreased in vivo following XTSJ treatment. CONCLUSION: XTSJ prevented GC metastasis by inhibiting the GnT-V-mediated E-cadherin glycosylation and promoting the E-cadherin accumulation at cell-cell junctions. Please cite this article as: Huang N, He HW, He YY, Gu W, Xu MJ, Liu L. Xiaotan Sanjie recipe, a compound Chinese herbal medicine, inhibits gastric cancer metastasis by regulating GnT-V-mediated E-cadherin glycosylation. J Integr Med. 2023; 21(6): 561-574.

Effects of polymer terminal group inside micelle core on paclitaxel loading promoting and burst release suppressing.

Background: Paclitaxel (PTX) is widely used in the treatment of advanced esophageal and gastric cancer. Polymeric micelles can improve the drug-loading efficiency of PTX. However, the end groups on the amphiphilic blocks affect the drug-loading efficiency and the release kinetics of polymeric micelles. Therefore, there is an urgent need to disclose the tailoring of the core-/shell-forming terminal groups. Methods: Different from the conventional block copolymer synthesis in the reversible addition-fragmentation chain-transfer polymerization, which has a hydrophilic end group on the core-forming blocks, an alternative monomer addition method was applied to tune and obtain two block copolymers with symmetrical and similar block length PBMAn-b-PNAMm [PNAM, poly(N-acryloylmorpholine); PBMA, poly(n-butyl methacrylate)] but distinct end groups on the hydrophobic core-forming blocks, that is, HOOC-PBMA-PNAM-Phen and HOOC-PNAM-PBMA-Phen. The chemical structure of the resulting copolymers was elucidated by proton nuclear magnetic resonance spectroscopy and differential scanning calorimetry. The spherical morphology revealed by transmission electron microscopy and the uniform particle size revealed by dynamic light scattering analysis clearly confirmed the successful preparation of a PTX-polymeric micelle complex. Results: The particle sizes of HOOC-PBMA-PNAM-Phen and HOOC-PNAM-PBMA-Phen were about 40 and 235 nm respectively. The PTX loading efficiency of HOOC-PBMA-PNAM-Phen was much lower than that of HOOC-PNAM-PBMA-Phen. The PTX release from HOOC-PBMA-PNAM-Phen was much slower than that of HOOC-PNAM-PBMA-Phen. The polymers had glass transition temperature (Tg) values of 70.24 and 74.22 ℃, which was from the HOOC-PBMA-PNAM-Phen and HOOC-PNAM-PBMA-Phen micelles, respectively. The systematic study on the PTX loading and releasing profile disclosed that, compared with the HOOC-PBMA-PNAM-Phen, the micelles with Phen group on the hydrophobic block (HOOC-PNAM-PBMA-Phen) enhanced drug loading and prolonged drug release but with a larger particle size. Conclusions: The results indicated that the hydrophobic end group Phen on the core-forming blocks can promote hydrophobic drug loading and suppress burst release.

Association between android fat mass, gynoid fat mass and cardiovascular and all-cause mortality in adults: NHANES 2003-2007.

Objectives: Evidence of the relationship between android fat mass and gynoid fat mass with the mortality prediction is still limited. Current study analyzed the NHANES database to investigate the relationship between android fat mass, gynoid fat mass and CVD, with all-cause mortality. Method: The study subjects were NHANES participants over 20 years old, two indicators of regional body composition, android fat and gynoid fat were measured by Dual Energy x-ray Absorptiometry (DEXA). The other various covariates data obtained from the NHANES questionnaire and laboratory measurements, including age, gender, education, race/ethnicity, uric acid, total serum cholesterol, albumin, Vitamin C, folate, alcohol drinking, smoking status, history of diabetes, and hypertension. Mortality status was ascertained from a linked mortality file prepared by the National Center for Health Statistics. The study population was divided quartiles based on the distribution of android fat mass and gynoid fat mass. The relationship between these two indicators with cardiovascular and all-cause mortality was investigated by using Cox regression. The covariates age, gender, smoking status, drinking status, history of diabetes, and history of hypertension were stratified. Results: In the fully adjusted model, Q3 had the lowest HR in android fat mass and gynoid fat mass. When examining the relationship between android fat mass and CVD mortality, current smokers and drinkers had the lowest CVD risk in Q2 [smoking: 0.21 (0.08, 0.52), drinking: 0.14 (0.04, 0.50)]. In diabetic patients, compared with Q1, other groups with increased android fat mass can significantly reduce the risk of CVD [Q4: 0.17 (0.04, 0.75), Q3: 0.18 (0.03, 1.09), Q2: 0.27 (0.09, 0.83)]. In ≥60 years old and female, the greater the gynoid fat mass, the smaller the HR of all-cause mortality [Q4 for ≥60 years old: 0.57 (0.33, 0.96), Q4 for female: 0.37 (0.23, 0.58)]. People <60 years old had a lower risk of all-cause mortality with gynoid fat mass in Q3 than those ≥60 years old [<60 years: 0.50 (0.27, 0.91), ≥60 years: 0.65 (0.45, 0.95)]. Among subjects without hypertension, the group with the largest android fat mass had the lowest risk of CVD mortality, and the group with the largest gynoid fat mass had the lowest risk of all-cause mortality [Android fat mass: 0.36 (0.16, 0.81), gynoid fat mass: 0.57 (0.39, 0.85)]. Conclusion: Moderate android fat mass and gynoid fat mass (Q3) had the most protective effect. Smokers and drinkers need to control their body fat. Being too thin is harmful to people with diabetes. Increased gynoid fat mass is a protective factor for all-cause mortality in older adults and females. Young people's gynoid fat mass is more protective in the moderate range than older people's. If no high blood pressure exists, people with more android and gynoid fat mass have a lower risk of CVD or all-cause mortality.

Structure and Function of SNM1 Family Nucleases.

Three human nucleases, SNM1A, SNM1B/Apollo, and SNM1C/Artemis, belong to the SNM1 gene family. These nucleases are involved in various cellular functions, including homologous recombination, nonhomologous end-joining, cell cycle regulation, and telomere maintenance. These three proteins share a similar catalytic domain, which is characterized as a fused metallo-ß-lactamase and a CPSF-Artemis-SNM1-PSO2 domain. SNM1A and SNM1B/Apollo are exonucleases, whereas SNM1C/Artemis is an endonuclease. This review contains a summary of recent research on SNM1's cellular and biochemical functions, as well as structural biology studies. In addition, protein structure prediction by the artificial intelligence program AlphaFold provides a different view of the proteins' non-catalytic domain features, which may be used in combination with current results from X-ray crystallography and cryo-EM to understand their mechanism more clearly.

Comprehensive evaluation of military training-induced fatigue among soldiers in China: A Delphi consensus study.

Objective: Military training-induced fatigue (MIF) often results into non-combat attrition. However, standard evaluation of MIF is unavailable. This study aimed to provide credible suggestions about MIF-evaluation. Methods: A 3-round Delphi study was performed. The authority of the experts was assessed by the authority coefficient (Aa). In round 1, categories of indicators were collected via anonymous survey of experts, then potential indicators were selected via literature search. In round 2, experts should evaluate the clinical implication, practical value, and importance of each potential indicators, or recommend new indicators based on feedback of round 1. Indicators with recommendation proportions ≥ 70% and new recommended indicators would be included in round 3 to be rated on a 5-point Likert scale. "Consensus in" was achieved when coefficient of concordance (Kendall's W) of a round was between 0.2 and 0.5 and the coefficient of variation (CV) of each aspect for an indicator was < 0.5. If round 3 could not achieve "consensus in," more rounds would be conducted iteratively based on round 3. Indicators included in the recommendation set were ultimately classified into grade I (highly recommended) or grade II (recommended) according to the mean score and CV of the aspects. Results: Twenty-three experts participated with credible authority coefficient (mean Aa = 0.733). "Consensus in" was achieved in round 3 (Kendall's W = 0.435, p < 0.001; all CV < 0.5). Round 1 recommended 10 categories with 73 indicators identified from 2,971 articles. After 3-round consultation, consensus was reached on 28 indicators focusing on the cardiovascular system (n = 4), oxygen transport system (n = 5), energy metabolism/metabolite level (n = 6), muscle/tissue damage level (n = 3), neurological function (n = 2), neuropsychological/psychological function (n = 3), endocrine function (n = 3), and exercise capacity (n = 2). Among these, 11 indicators were recommended as grade I: basic heart rate, heart-rate recovery time, heart rate variability, hemoglobin, blood lactic acid, urine protein, creatine kinase, reaction time, Borg Rating of Perceived Exertion Scale, testosterone/cortisol, and vertical jump height. Conclusion: This study developed a reliable foundation for the comprehensive evaluation of MIF among soldiers.

Association of lung function with the risk of cardiovascular diseases and all-cause mortality in patients with diabetes: Results from NHANES III 1988-1994.

Objective: The potential effects of pulmonary dysfunction on cardiovascular diseases (CVD) and all-cause mortality are receiving attention. The current study aimed to explore whether reduced lung function predicts CVD and all-cause mortality in people with diabetes. Methods: A total of 1,723 adults with diabetes (mean age 60.2 years) were included in the National Health and Nutrition Examination Survey (NHANES III). Death outcomes were ascertained by linkage to the database records through 31 December 2015. Cox proportional hazards regression models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for coronary heart disease (CHD), CVD, and all-cause mortalities. We conducted stratified analyses based on age, body mass index (BMI), history of hypertension, and dyslipidemia. Results: During a mean follow-up of 14.62 years (25,184 person-year), a total of 1,221 deaths were documented, of which 327 were CHD, 406 were CVD, and 197 were cancer. After multi-factor adjustment, participants with lower FEV1 and FVC had a higher risk of CHD, CVD, and all-cause mortality. This association was also found in lower FVC and a higher risk of cancer mortality [HR: 3.85 (1.31-11.32); P for trend = 0.040], but the association of FEV1 was attenuated after adjustment for covariates [HR:2.23 (0.54-9.17); P for trend = 0.247]. In subgroup analysis, we found that the adverse associations of FEV1 and FVC with CVD mortality were observed in subgroups of age, BMI, and history of hypertension and dyslipidemia. Conclusion: Declined lung function was associated with a higher risk of CVD and all-cause mortality in people with diabetes. Lung function tests, especially FEV1 and FVC, should be encouraged to provide prognostic and predictive information for the management of CVD and all-cause mortality in patients with diabetes.

MiR-200c-3p and miR-485-5p overexpression elevates cisplatin sensitivity and suppresses the malignant phenotypes of non-small cell lung cancer cells through targeting RRM2.

BACKGROUND: This study intended to investigate the potential mechanism of microRNA-200c-3p (miR-200c-3p) and miR-485-5p in mediating the cisplatin (DDP) resistance in non-small cell lung cancer (NSCLC). METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to measure the expression of miR-200c-3p, miR-485-5p, and ribonucleotide reductase regulatory subunit M2 (RRM2) messenger RNA (mRNA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to analyze the DDP resistance and the proliferation of NSCLC cells. Colony formation assay was used to assess cell proliferation. Transwell assays were used to evaluate cell migration and invasion. The target relationship between RRM2 and miR-200c-3p or miR-485-5p was verified using dual-luciferase reporter assay. The protein level of RRM2 was measured using Western blot assay. Animal experiments were conducted to analyze the roles of miR-200c-3p and miR-485-5p in the DDP resistance of xenograft tumors in vivo. RESULTS: MiR-200c-3p and miR-485-5p were both downregulated in DDP-resistant NSCLC tissues and cell lines. Overexpressing miR-200c-3p or miR-485-5p suppressed the DDP resistance and malignant behaviors of NSCLC cells. MiR-200c-3p played a synergistic role with miR-485-5p in regulating the chemo-resistance and biological behaviors NSCLC cells. RRM2 was confirmed as a target of miR-200c-3p and miR-485-5p. RRM2 silencing restrained the DDP resistance and progression of NSCLC. RRM2 overexpression partly reversed miR-200c-3p or miR-485-5p-induced influences in NSCLC cells. The overexpression of miR-200c-3p or miR-485-5p aggravated DDP-mediated suppressive effect on tumor growth in vivo. CONCLUSION: MiR-200c-3p or miR-485-5p enhanced the DDP sensitivity and suppressed the malignant behaviors of NSCLC cells partly through targeting RRM2.

Efficacy of the Synergy Between Live-Attenuated and Inactivated PRRSV Vaccines Against a NADC30-Like Strain of Porcine Reproductive and Respiratory Syndrome Virus in 4-Week Piglets.

The NADC30-like strain of porcine reproductive and respiratory syndrome virus (PRRSV) is a novel strain responsible for substantial economic losses to swine production in China. This study evaluated the cross-protective efficacy of the synergy between live-attenuated and inactivated PRRSV vaccines compared with a single vaccination with PRRS modified-live virus (MLV) vaccine against challenge with NADC30-like strain, v2016/ZJ/09-03. A total of 45 PRRSV free pigs were randomly divided into five groups: (1) strict control (SC); (2) positive control (PC); (3) single MLV dose (M1); (4) primed intramuscularly with MLV and boosted with killed vaccine 3 weeks later (MK1); and (5) intramuscular prime MLV boosted subcutaneously with killed vaccine B 3 weeks later (MK2). Serological tests in MK groups revealed no differences in both anti-N and anti-GP protein antibodies compared with M1 group, and failed to provide further protection against clinical signs, virus shedding, and gross lesions. However, the viremic titer, gross lung lesions, and average daily weight gain were significantly improved in the MLV vaccinated groups, suggesting that MLV provides substantial cross-protection against the NADC30-like virus. Thus, as a booster, the killed vaccine confers minimal additional protection in NADC30-like infected piglets.

Core muscle functional strength training for reducing the risk of low back pain in military recruits: An open-label randomized controlled trial.

BACKGROUND: Core muscle functional strength training (CMFST) has been reported to reduce injuries to the lower extremity. However, no study has confirmed whether CMFST can reduce the risk of low back pain (LBP). OBJECTIVE: This study identified the effects of CMFST on the incidence of LBP in military recruits. DESIGN, SETTING, PARTICIPANTS AND INTERVENTION: We performed a prospective, open-label, randomized, controlled study in a population of young healthy male naval recruits from a Chinese basic combat training program. Participants were randomly assigned to either the core group or the control group. In additional to normal basic combat training, recruits in the core group underwent a CMFST program for 12 weeks, while recruits in the control group received no extra training. MAIN OUTCOME MEASURES: At the beginning of the study and at the 12th week, the number of participants with LBP was counted, and lumbar muscle endurance was measured. In addition, when participants complained of LBP, they were assessed using the visual analog scale (VAS) and Roland Morris Disability Questionnaire (RMDQ). RESULTS: A total of 588 participants were included in the final analysis (295 in the core group and 293 in the control group). The incidence of LBP in the control group was about twice that of the core group over the 12-week study (20.8% vs 10.8%, odds ratio: 2.161-2.159, P < 0.001). The core group had better lumbar muscle endurance at 12 weeks than the control group ([200.80 ± 92.98] s vs [147.00 ± 84.51] s, P < 0.01). There was no significant difference in VAS score between groups, but the core group had a significantly lower RMDQ score at week 12 than the control group (3.33 ± 0.58 vs 5.47 ± 4.41, P < 0.05). CONCLUSION: This study demonstrated that the CMFST effectively reduced the incidence of LBP, improved lumbar muscle endurance, and relieved the dysfunction of LBP during basic military training.

Associations of eicosapentaenoic acid and docosahexaenoic acid intakes with cardiovascular and all-cause mortality in patients with diabetes: Result from National Health and Nutrition Examination Survey 1999-2008.

Introduction: The evidence on eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) intake status and long-term mortality among people with diabetes is scarce. This study aimed to investigate the relationship between EPA and DHA intakes with all-cause and cause-specific mortality in adults with diabetes. Methods: This study included 2,991 adults with diabetes from the National Health and Nutrition Examination Survey (NHANES) 1999-2008. Death outcomes were ascertained by linkage to the database records through 31 December 2015. Cox proportional hazards regression models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for mortality from all causes, cardiovascular disease (CVD), and coronary heart disease (CHD) in patients with diabetes. Results: Among 2,991 patients with diabetes, the mean age was 61.9 years (55.2% males). During the mean follow-up duration of 9.4 years, a total of 1,091 deaths were documented, of which 273 were due to CVD, including 227 CHD deaths. EPA and DHA intakes were associated with lower mortality risks, especially that of CVD. After adjusting for demographic, major lifestyle factors, overall dietary intake patterns, and history of hypertension and dyslipidemia, the multivariable HRs (95% CIs) of mortality risk comparing Q4 to Q1 of EPA intake were 0.55 (0.33-0.92; P-trend = 0.019) for CHD, 0.55 (0.36-0.83; P-trend = 0.005) for CVD, and 0.91 (0.70-1.18; P-trend = 0.264) for all-cause. The respective HRs (95% CIs) comparing Q4 to Q1 of DHA were 0.60 (0.37-0.98; P-trend = 0.051) for CHD, 0.58 (0.38-0.89; P-trend = 0.014) for CVD, and 0.92 (0.72-1.18; P-trend = 0.481) for all-cause. In subgroup analysis, we found that the association trends of EPA and DHA intakes with death risk remained robust among patients with diabetes, especially among those who are old, female, those with higher BMI, and dyslipidemia patients with CVD and CHD. Discussion: In the USA, higher EPA and DHA intakes were associated with a lower risk of CHD and CVD mortality in patients with diabetes. Our study supports the benefits of adequate EPA and DHA intakes in promoting the health of patients with diabetes.

A systematic review and meta-analysis of acupuncture combined with Tuina in the treatment of insomnia.

BACKGROUND: Insomnia is a sleep disorder with insufficient sleep time or/and poor sleep quality. Relevant epidemiological studies have shown that insomnia symptoms occur in about 35% to 50% of the adult population, and it is one of the most common diseases in the elderly. Patients who often suffer from insomnia are prone to symptoms such as fatigue, weakened cognitive function, depression, and even mental illness, which bring serious physical and mental damage to individuals and a heavy economic burden to social medical care and families. Traditional Chinese medicine and modern medicine have their own advantages in the treatment of insomnia, and there is currently a lack of reports on the comparison of acupuncture combined with massage and conventional medicine. OBJECTIVE: To evaluate the clinical efficacy of acupuncture combined with Tuina in the treatment of insomnia. METHODS: Search for clinical randomized controlled trials (RCTs) of acupuncture combined with Tuina in the treatment of insomnia from PubMed, Cochrane Library, Web of Science, China National Knowledge Infrastructure, Wan Fang Database, and China Science and Technology Journal Database. The RevMan5.4 software was used for Meta- analysis after literature screening, data extraction and quality evaluation. RESULTS: A total of 29 studies were included with a total of 2688 cases. Compared with drugs or acupuncture alone, acupuncture combined with Tuina has advantages in the total clinical effectiveness, as well as the Pittsburgh Sleep Quality Index (PSQI) and Statistical Self-Rating Anxiety Scale score (SAS) (OR = 3.59, 95% confidence interval [CI] [2.77, 4.66], Z = 9.62 [P < .00001]) (MD = -2.44, 95% CI [-2.93, -1.95], Z = 9.72 [P < .00001]) (MD = -8.42, 95% CI [-10.23, -6.61], Z = 9.09 [P < .00001]). There was no statistically significant difference in Statistical Self-rating Depression Scale score (SDS) (MD = -5.26, 95% CI [-11.29, 0.78], Z = 1.71 [P > .05]). CONCLUSION: Acupuncture combined with Tuina has obvious clinical advantages in the treatment of insomnia. This result is expected to provide a reference for the clinical treatment of insomnia, but the long-term effect of clinical efficacy still needs further study.

Efficient and Scalable Production of Full-length Human Huntingtin Variants in Mammalian Cells using a Transient Expression System.

Full-length huntingtin (FL HTT) is a large (aa 1-3,144), ubiquitously expressed, polyglutamine (polyQ)-containing protein with a mass of approximately 350 kDa. While the cellular function of FL HTT is not entirely understood, a mutant expansion of the polyQ tract above ~36 repeats is associated with Huntington's disease (HD), with the polyQ length correlating roughly with the age of onset. To better understand the effect of structure on the function of mutant HTT (mHTT), large quantities of the protein are required. Submilligram production of FL HTT in mammalian cells was achieved using doxycycline-inducible stable cell line expression. However, protein production from stable cell lines has limitations that can be overcome with transient transfection methods. This paper presents a robust method for low-milligram quantity production of FL HTT and its variants from codon-optimized plasmids by transient transfection using polyethylenimine (PEI). The method is scalable (>10 mg) and consistently yields 1-2 mg/L of cell culture of highly purified FL HTT. Consistent with previous reports, the purified solution state of FL HTT was found to be highly dynamic; the protein has a propensity to form dimers and high-order oligomers. A key to slowing oligomer formation is working quickly to isolate the monomeric fractions from the dimeric and high-order oligomeric fractions during size exclusion chromatography. Size exclusion chromatography with multiangle light scattering (SEC-MALS) was used to analyze the dimer and higher-order oligomeric content of purified HTT. No correlation was observed between FL HTT polyQ length (Q23, Q48, and Q73) and oligomer content. The exon1-deleted construct (aa 91-3,144) showed comparable oligomerization propensity to FL HTT (aa 1-3,144). Production, purification, and characterization methods by SEC/MALS-refractive index (RI), sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), western blot, Native PAGE, and Blue Native PAGE are described herein.

Intrathecal IGF2 siRNA injection provides long-lasting anti-allodynic effect in a spared nerve injury rat model of neuropathic pain.

Previous studies have shown an increase of insulin-like growth factor-2 (IGF2) in animal models of neuropathic pain. We aimed to examine the hypothesis that reducing the expression of IGF2 using intrathecal IGF2 small-interfering RNA (siRNA) would attenuate the development of neuropathic pain in rats after spared nerve injury (SNI). Male Wistar rats were divided into three groups: sham-operated group, in which surgery was performed to cut the muscles without injuring the nerves; SNI group, in which SNI surgery was performed to sever the nerves; and SNI + siRNA IGF2 group, in which SNI surgery was performed, and IGF2-siRNA was administered intrathecally 1 day after SNI. The rats were assessed for mechanical allodynia and cold allodynia 1 day before surgery (baseline), and at 2, 4, 6, 8, and 10 days after siRNA treatment. The rat spinal cord was collected for quantitative polymerase chain reaction and western blot analysis. Compared with the SNI group, rats that received IGF2 siRNA showed a significantly increased SNI-induced paw-withdrawal threshold to metal filament stimulation from Day 4 to Day 10 after SNI surgery. IGF2 siRNA significantly decreased the response duration from the acetone test from Day 2 to Day 10 following SNI surgery. SNI increased IGF2 mRNA expression on Day 2 and increased IGF2 protein expression on Day 8 and Day 10 in the spinal cord of the SNI rats. However, the above-mentioned effects of IGF2 mRNA and protein expression were significantly inhibited in the SNI + IGF2 siRNA group. We demonstrated that intrathecal administration of IGF2 siRNA provided significant inhibition of SNI-induced neuropathic pain via inhibition of IGF2 expression in the spinal cord. The analgesic effect lasted for 10 days. Further exploration of intrathecal IGF2 siRNA administration as a potential therapeutic strategy for neuropathic pain is warranted.

Detection and Evaluation of Serological Biomarkers to Predict Osteoarthritis in Anterior Cruciate Ligament Transection Combined Medial Meniscectomy Rat Model.

Biomarkers are essential tools in osteoarthritis (OA) research, clinical trials, and drug development. Detecting and evaluating biomarkers in OA research can open new avenues for researching and developing new therapeutics. In the present report, we have explored the serological detection of various osteoarthritis-related biomarkers in the preclinical model of OA. In this surgical OA model, we disrupted the medial tibial cartilage's integrity via anterior cruciate ligament transection combined with medial meniscectomy (ACLT+MMx) of a single joint of Wistar rats. The progression of OA was verified, as shown by the microscopic deterioration of cartilage and the increasing cartilage degeneration scoring from 4 to 12 weeks postsurgery. The concentration of serological biomarkers was measured at two timepoints, along with the complete blood count and bone electrolytes, with biochemical analysis further conducted. The panel evaluated inflammatory biomarkers, bone/cartilage biomarkers, and lipid metabolic pathway biomarkers. In chronic OA rats, we found a significant reduction of total vitamin D3 and C-telopeptide fragments of type II (CTX-II) levels in the serum as compared to sham-operated rats. In contrast, the serological levels of adiponectin, leptin, and matrix metallopeptidase (MMP3) were significantly enhanced in chronic OA rats. The inflammatory markers, blood cell composition, and biochemical profile remained unchanged after surgery. In conclusion, we found that a preclinical model of single-joint OA with significant deterioration of the cartilage can lead to serological changes to the cartilage and metabolic-related biomarkers without alteration of the systemic blood and biochemical profile. Thus, this biomarker profile provides a new tool for diagnostic/therapeutic assessment in OA scientific research.

Postoperative Drip-Infusion of Remifentanil Reduces Postoperative Pain-A Retrospective Observative Study.

Development of remifentanil-induced hyperalgesia (RIH) postoperatively is an unpleasant experience that requires further treatment. This study assessed the effects of gradual withdrawal combined with drip infusion of remifentanil on postoperative pain and the requirement for rescue analgesics. A total of 559 patients receiving total intravenous anesthesia with propofol and remifentanil were enrolled. All patients either underwent gradual withdrawal of remifentanil (GWR) or gradual withdrawal combined with drip infusion (GWDR) with a dose of 1 mcg·kg-1 for 30 min after extubation. The numeric rating scale (NRS) and the requirement of rescue analgesics were assessed. The requirement for rescue analgesics was significantly lower in the GWDR group than in the GWR group (13.2% vs. 35.7%; p < 0.001). At the post-anesthetic care unit (PACU), patients in the GWDR group had a lower NRS pain score (p < 0.001). In addition, in the postoperative 2nd hour, patients in the GWDR group had a significantly lower NRS than the GWR group (beta, -0.31; p = 0.003). No remifentanil-related adverse effects were observed. We found that gradual withdrawal combined with drip infusion of remifentanil required less rescue analgesics and reduced pain scores. The new way of remifentanil administration may be effective to prevent RIH.

Inhibition of leucine-rich repeats and calponin homology domain containing 1 accelerates microglia-mediated neuroinflammation in a rat traumatic spinal cord injury model.

BACKGROUND: Spinal cord injury (SCI) triggers the primary mechanical injury and secondary inflammation-mediated injury. Neuroinflammation-mediated insult causes secondary and extensive neurological damage after SCI. Microglia play a pivotal role in the initiation and progression of post-SCI neuroinflammation. METHODS: To elucidate the significance of LRCH1 to microglial functions, we applied lentivirus-induced LRCH1 knockdown in primary microglia culture and tested the role of LRCH1 in microglia-mediated inflammatory reaction both in vitro and in a rat SCI model. RESULTS: We found that LRCH1 was downregulated in microglia after traumatic SCI. LRCH1 knockdown increased the production of pro-inflammatory cytokines such as IL-1ß, TNF-α, and IL-6 after in vitro priming with lipopolysaccharide and adenosine triphosphate. Furthermore, LRCH1 knockdown promoted the priming-induced microglial polarization towards the pro-inflammatory inducible nitric oxide synthase (iNOS)-expressing microglia. LRCH1 knockdown also enhanced microglia-mediated N27 neuron death after priming. Further analysis revealed that LRCH1 knockdown increased priming-induced activation of p38 mitogen-activated protein kinase (MAPK) and Erk1/2 signaling, which are crucial to the inflammatory response of microglia. When LRCH1-knockdown microglia were adoptively injected into rat spinal cords, they enhanced post-SCI production of pro-inflammatory cytokines, increased SCI-induced recruitment of leukocytes, aggravated SCI-induced tissue damage and neuronal death, and worsened the locomotor function. CONCLUSION: Our study reveals for the first time that LRCH1 serves as a negative regulator of microglia-mediated neuroinflammation after SCI and provides clues for developing novel therapeutic approaches against SCI.

Propofol-based total intravenous anesthesia improves survival compared to desflurane anesthesia in gastric cancer surgery: A retrospective analysis.

Surgical management of cancer may induce stress and increase the likelihood of cancer metastasis and recurrence. Appropriate surgical and anesthetic techniques may affect the patient's outcome. Although numerous studies have been performed, conflicting results have been obtained regarding the effect of anesthetic techniques on the outcome of patients with cancer. We conducted this study to evaluate the association of anesthetic techniques with overall and recurrence-free survival in patients who had undergone gastric cancer surgery.This retrospective study reviewed the electronic medical records of patients, who had visited our hospital and had been diagnosed with gastric cancer between July 1st, 2006 to June 30th, 2016. Univariate analysis of the potential prognostic factors was performed using the log-rank test for categorical factors, and parameters with a P-value < .05 at the univariate step were included in the multivariate regression analysis. Propensity Score Matching was performed to account for differences in baseline characteristics: propofol or desflurane, in a 1:1 ratio.A total of 408 patients anesthetized with desflurane (218) and propofol (190) were eligible for analysis. After propensity matching, 167 patients remained in each group. The overall mortality rate was significantly higher in the desflurane group (56%) than in the propofol group (34%) during follow-up (P < .001). In addition, a greater percentage of patients in the desflurane group (41%) exhibited postoperative metastasis than those in the propofol group (19%, P < .001).The authors found some association between types of anesthesia used and the long-term prognosis of gastric cancer. Propofol-based total intravenous anesthesia improved survival and reduced the risk of recurrence and metastasis during the 5-year follow-up period after gastric cancer surgery.

Structural analysis of the catalytic domain of Artemis endonuclease/SNM1C reveals distinct structural features.

The endonuclease Artemis is responsible for opening DNA hairpins during V(D)J recombination and for processing a subset of pathological DNA double-strand breaks. Artemis is an attractive target for the development of therapeutics to manage various B cell and T cell tumors, because failure to open DNA hairpins and accumulation of chromosomal breaks may reduce the proliferation and viability of pre-T and pre-B cell derivatives. However, structure-based drug discovery of specific Artemis inhibitors has been hampered by a lack of crystal structures. Here, we report the structure of the catalytic domain of recombinant human Artemis. The catalytic domain displayed a polypeptide fold similar overall to those of other members in the DNA cross-link repair gene SNM1 family and in mRNA 3'-end-processing endonuclease CPSF-73, containing metallo-ß-lactamase and ß-CASP domains and a cluster of conserved histidine and aspartate residues capable of binding two metal atoms in the catalytic site. As in SNM1A, only one zinc ion was located in the Artemis active site. However, Artemis displayed several unique features. Unlike in other members of this enzyme class, a second zinc ion was present in the ß-CASP domain that leads to structural reorientation of the putative DNA-binding surface and extends the substrate-binding pocket to a new pocket, pocket III. Moreover, the substrate-binding surface exhibited a dominant and extensive positive charge distribution compared with that in the structures of SNM1A and SNM1B, presumably because of the structurally distinct DNA substrate of Artemis. The structural features identified here may provide opportunities for designing selective Artemis inhibitors.

A T7 exonuclease assisted dual-cycle signal amplification assay of miRNA using nanospheres-enhanced fluorescence polarization.

Based on streptavidin coated nanospheres and T7 exonuclease assisted dual-cycle signal amplification, we developed a novel sensitive fluorescence polarization detection method for miRNA. When target miRNA was present in the system, hairpin probe hybridized with miRNA, forming a double-stranded structure. The 5' end of hairpin probe was then recognized and digested by T7 exonuclease, releasing the non-degraded single strand DNA fragments and miRNA. The released target miRNA could trigger the next cycle of hybridization and digestion, releasing more non-degraded fragments from hairpin probe. The fragments could hybridize with a signal probe (with carboxyfluorescein modification at 5'-end and biotin modification at 3'-end). The formed blunt 5'-end of signal probe was then recognized and degraded by T7 exonuclease, releasing the fragments and the fluorophore carboxyfluorescein. The next cycle of hybridization and digestion of signal probe was triggered by the released fragment at the same time. The free carboxyfluorescein cannot connect with streptavidin coated nanospheres which were used as the fluorescence polarization signal amplifier. So, there was a big change of fluorescence polarization signal after adding miRNA into the detection system, due to the different fluorescence polarization signal between nanospheres-captured intact signal probe and free carboxyfluorescein. The detection limit of this method is about 0.001 nM, and it has a good selectivity. In addition, it was also applicable to determine target miRNA in total miRNA extracts and compare the expression level of target miRNA in different cells. Consequently, the proposed method is expected to be used for the potential cancer diagnosis and the related biomedical research.