Determination and difference analysis of bile acid active constituents of artificial bezoar in Babao jingfeng powder from different manufacturers / 中国药房

OBJECTIVE To establish the method for determination of bile acid active constituents （cholic acid， hyodeoxycholic acid and sodium taurocholate） in Babao jingfeng powder， and to compare the differences of 3 constituents in Babao jingfeng powder from different manufacturers. METHODS UPLC-MS method was adopted to determine the contents of cholic acid， hyodeoxycholic acid and sodium taurocholate in 9 batches of Babao jingfeng powder from 3 manufacturers. The separation was performed on a Phenomenex Luna® C18 column with mobile phase consisted of acetonitrile-water （gradient elution） at a flow rate of 0.5 mL/min. The column temperature was 40 ℃ and the sample size was 2 μL. MS system was operated in a negative ion single quadrupole mode， and the m/z of cholic acid， hyodeoxycholic acid and sodium taurocholate were 407.2， 392.2 and 514.2， respectively. With the contents of three components as the index， cluster analysis and principle component analysis were performed on 9 batches of samples. RESULTS The linear ranges of cholic acid， hyodeoxycholic acid and sodium taurocholate were 5.48- 548.40， 5.38-538.40， 4.74-474.05 ng/mL， respectively （r≥ 0.999 3）. RSDs of precision， repeatability and stability tests（24 h） were all lower than or equal to 3.7% （n＝6）， and the average recoveries were 103.3%， 103.3% and 101.6% with RSDs of 3.3%， 3.4%， 4.2% （n＝6）， respectively. The contentsof cholic acid ranged 0.702-1.711 mg/g， those of deoxycholic acid ranged 0.599-2.049 mg/g， and those of sodium taurocholate ranged 0.664-1.752 mg/g in 9 batches of samples. The average content of 3 components in samples from manufacturer A was high， and the difference between batches was the smallest； the average content of 3 components from manufacturer C was the lowest， and the difference between batches was large. Cluster analysis could basically distinguish samples from different manufacturers； cluster analysis was conducted by using the comprehensive score of principal components as an indicator， the results of which were basically consistent with those of cluster analysis for content. CONCLUSIONS The method is established successfully for the content determination of the 3 bile acid active constituents of artificial bezoar in Babao jingfeng powder. The contents of 3 components in Babao jingfeng powder from 3 manufacturers are significantly different.

Research progress of Kelch-like ECH-associated protein 1-nuclear factor erythroid 2-related factor 2 signaling pathway regulated retinal oxidative stress / 国际眼科杂志(Guoji Yanke Zazhi)

Oxidative stress(OS)is a major reason for body damage. Studies have shown that a variety of factors, such as ischemia and hypoxia, excessive light and hyperglycemia can cause the increase of reactive oxygen species and free radicals in the retina, thus inducing OS, damaging retina and affecting the normal visual function. Kelch-like ECH-associated protein 1(KEAP1)and nuclear factor erythroid 2 related factor 2(NRF2), which together constitute the main antioxidant stress signaling pathway in the body, play an antioxidant role by regulating retinal energy metabolism and cell proliferation, apoptosis and autophagy through various ways, so as to reduce retinal damage caused by OS. In this paper, the role and mechanism of the KEAP1-NRF2 signaling pathway regulation of OS in the retinal are briefly reviewed, aiming to provide ideas for subsequent research.

Effects of low-energy red light on choroidal thickness and spherical equivalent and axial length in myopic children / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To systematically evaluate the effects of low-energy red light on choroidal thickness(ChT), spherical equivalent(SE), and axial length(AL)in myopic children.METHODS: A total of 8 databases, including Pubmed, Embase, Cochrane Library, Web of Science, CNKI, WanFang Database, VIP Database and China Biomedical Literature Database, were electronically retrieved to collect the clinical randomized controlled trial(RCT)of low-energy red light in myopia, with red light intervention as an experimental group and glasses as a control group. The retrieval time limit is from the establishment of the database to January 2023. According to the recommendation of the Cochrane Handbook, literature quality and risk of bias were evaluated. Revman5.4 software was used for Meta-analysis.RESULTS: Totally 8 RCT about red-light treatment with myopia were included. The sample size for ChT analysis contained 407 eyes in the red-light group and 425 eyes in the control group; SE analysis included 490 eyes in the red-light group and 518 eyes in the control group; sample size for AL analysis were 518 eyes in the red-light group and 539 eyes in the control group. The differences in ChT, SE and AL between the groups were statistically significant(ChT: WMD=37.81, 95%CI: 21.55～54.07, P<0.001; SE: WMD=0.58, 95%CI: 0.46～0.71, P<0.001; AL: WMD=-0.31, 95%CI: -0.39～-0.24, P<0.001).CONCLUSION: Specific red light can promote the increase of ChT while inhibit SE and AL. However, its long-term efficacy and side effects remain unclear. The above conclusions need to be further clarified by more RCT with large samples.

Clinical efficacy of temozolomide combined with X-ray stereotactic radiotherapy in the treatment of recurrent glioma / 中国医师杂志

Objective:To investigate the curative efficacy of X-ray stereotactic radiotherapy combined with temozolomide in the treatment of recurrent glioma.Methods:48 patients with recurrent glioma treated in Mianyang Central Hospital from January 2018 to January 2019 were retrospectively selected, including 24 patients treated with stereotactic radiotherapy as the control group and 24 patients treated with temozolomide combined with stereotactic radiotherapy as the observation group. The treatment effect, inflammatory factor level, incidence of adverse events and survival rate were compared between the two groups.Results:The complete remission rate and total effective rate in the observation group were significantly higher than those in the control group (66.7% vs 37.5%, 87.5% vs 62.5%) (all P<0.05). There were no significant differences in serum levels of hepatocyte growth factor (HGF), tumor necrosis factor-α (TNF-α) and interleukin-17 (IL-17) between the two groups before treatment (all P>0.05). After treatment, the serum levels of HGF, TNF-α and IL-17 in observation group was significantly lower than those in control group (all P<0.05). The incidence of adverse events in the observation group was significantly lower than that in the control group ( P<0.05). During follow-up of 6, 12 and 18 months, the survival rate of patients in the observation group was significantly higher than that in the control group, with statistically significant difference (all P<0.05). Conclusions:The combination of X-ray stereotactic radiotherapy with temozolomide in the treatment of recurrent glioma shows better clinical outcome and extended survival rate. To conclude, this combined treatment is recommended in further clinical promotion.

Role of Shh/Gli1 signaling pathway in sleep deprivation-induced cognitive impairment in young mice / 中华麻醉学杂志

Objective:To evaluate the role of sonic hedgehog (Shh)/glioma-associated oncogene homolog 1 (Gli1) signaling pathway in sleep deprivation-induced cognitive impairment in young mice.Methods:Forty-eight SPF healthy male C57BL/6 mice, aged 4 weeks, weighing 14-16 g, were divided into 3 groups ( n=16 each) by the random number table method: control group (C group), sleep deprivation group (SD group) and Shh agonist SAG group (SD+ SAG group). Multi-platform water environment method was used to prepare the sleep deprivation model in mice, and the sleep deprivation was 20 h a day for 10 consecutive days.In SD+ SAG group, SAG 10 mg/kg was intraperitoneally injected at 5 min before each sleep deprivation, while the equal volume of normal saline was intraperitoneally injected in group C and group SD.The mice underwent novel object recognition and Y-maze tests at 24 h after development of the model.Mice were sacrificed after the behavioral testing, and the hippocampi were isolated for determination of the density of dendritic spines in hippocampal CA1 region (by Golgi staining), expression of Gli1 and brain-derived neurotrophic factor (BDNF) in hippocampal tissues (by Western blot), and expression of Gli1 and BDNF mRNA in hippocampal tissues (by quantitative real-time polymerase chain reaction). Results:Compared with group C, the preference index in novel object recognition and Y-maze tests and density of dendritic spines in CA1 region were significantly decreased, and the expression of Gli1 and BDNF protein and mRNA in hippocampus was down-regulated in group SD ( P<0.05). Compared with group SD, the preference index in novel object recognition and Y-maze tests and density of dendritic spines in CA1 region were significantly increased, and the expression of Gli1 and BDNF protein and mRNA in hippocampus was up-regulated in group SD+ SAG ( P<0.05). Conclusions:Inhibition of Shh/Gli1 signaling pathway and reduction of plasticity of dendritic spines of hippocampal neurons are involved in sleep deprivation-induced cognitive impairment in young mice.

Relationship between thalamocortical glutamate and neuronal activity in mice with sleep disorders induced by neuropathic pain / 中华麻醉学杂志

Objective:To evaluate the relationship between thalamocortical glutamate and neuronal activity in mice with neuropathic pain-induced sleep disorders.Methods:SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 15-25 g, were divided into 2 groups ( n=14 each) using a random number table method: sham operation group (Sham group) and neuropathic pain group (CCI group). Neuropathic pain was induced by chronic constrictive injury (CCI). The animals were anesthetized with intraperitoneal 10% chloral hydrate 3 ml/kg.The right sciatic nerve was exposed and 4 ligatures were placed on the sciatic nerve at 1 mm intervals.The mechanical paw withdrawal threshold and thermal paw withdrawal latency on the operated side were measured at 1 day before CCI (T 0) and 3, 5, 7, 14 and 21 days after CCI (T 1-5). Electroencephalogram recording electrodes were stereotaxically implanted in visual cortex at T 3, and electroencephalogram were monitored for 6 h, the percentages of non-rapid eye movement, rapid eye movement and wakefulness in the total time were calculated.Microwire electrodes were implanted epidurally over the ventral posterior (VP) nucleus of the thalamus and primary somatosensory cortex (S1) using a brain stereotaxic apparatus at T 3, and the data acquisition system was used to record field potentials at T 4, the percentage of power of each wave was calculated, and the coherence of the field potentials of VP and S1 was simultaneously evaluated.The mice were sacrificed at T 4, brain tissues were collected, and proton nuclear magnetic resonance spectroscopy was used to determine the level of neurotransmitter in the thalamus and cortex. Results:Compared with group Sham, the mechanical paw withdrawal threshold was significantly decreased and thermal paw withdrawal latency was shortened at T 1-5, the percentage of non-rapid eye movement time was decreased, the percentage of wakefulness time was increased, the percentage of δ wave power in the VP area was decreased, the percentage of δ wave power in the VP and S1 areas was increased, and the coherence of the field potentials of VP-S1 was increased in the frequency range of δ wave (1-4 Hz) and α wave (8-14 Hz), and the levels of glutamate, glutamine and glutamate-glutamine in the thalamus and cortex were increased in group CCI ( P<0.05). Conclusions:Neuropathic pain-induced sleep disturbance is related to increased thalamocortical glutamate levels, resulting in changes in the electrical activity of thalamocortical neurons of mice.

Effects of mesenchymal stem cells on primary culture and secretion function of parathyroid cells in vitro / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the effects of adipose-derived mesenchymal stem cells (ADMSCs) on proliferation and hormone secretion of parathyroid cells in votro. Methods: The parathyroid cells and ADMSCs were obtained from 10 SD rats by cell separation and culture. The phenotype of P3 generation for ADMSCs was detected by flow cytometry. The co-culture of parathyroid cells and ADMSCs was conducted in the ratios of 2∶1, 1∶1, 1∶2 and 1∶5, respectively. The level of parathyroid hormone in cell supernatant was determined. The results were compared with the parathyroid hormone in the supernatant of parathyroid cells cultured separately in the corresponding number. The effects of ADMSCs on the hormone secretion of parathyroid cells were evaluated. SPSS 11.0 software was used for statistical analysis. Results: The primary culture of either parathyroid cells or ADMSCs and the co-culture of these cells in vitro were performed successfully, and the in vitro culture of different proportions of the two cells showed different effects on parathyroid hormone secretion. The co-culture of parathyroid cells and ADMSCs, especially in the ratio of 1∶5, facilitated the secretion of parathyroid hormone ((1.3±0.0) vs. (0.8±0.1), (1.3±0.0) vs. (0.9±0.0), (1.7±0.5) vs. (0.9±0.0), (1.7±0.0) vs. (1.2±0.2))ng/L with t value of 25.46, 64.30, 3.32, 7.16, P<0.05 on the 2nd, 4th, 6th and 8th days respectively. Secondly, when the ratio was 1∶2, the PTH level showed an upward trend. Conclusion: Parathyroid cells and ADMSCs can be co-cultured in vitro, facilitating the secretion of parathyroid hormone under the appropriate cell proportion such as the ratio of by parathyroid cells to ADMSCs at 1∶5.

A new classification of maxillary defect and simultaneous accurate reconstruction / 中华耳鼻咽喉头颈外科杂志

Objective: To select the preferred flaps for the reconstruction of different maxillary defects and to propose a new classification of maxillary defects. Methods: A total of 219 patients (136 males and 83 females) underwent the simultaneous reconstruction of maxillary defects in the Beijing Tongren Hospital, Capital Medical University, between January 2005 and December 2018 were reviewed. Age ranged from 16 to 78 years. Based on the proposed new classification of the maxillary defects, 22 patients with class Ⅰ defects (inferior maxillectomy), 44 patients with class Ⅱ defects (supperior maxillectomy), 132 patients with class Ⅲ defects (total maxillectomy) and 21 patients with class Ⅳ defects (extensive maxillectomy) were enrolled. Survival rate, functional and aesthetic outcomes of flaps were evaluated. Survival analysis was performed in 169 patients with malignant tumor, Kaplan-Meier method was used to calculate the survival rate, and Log-rank method was used to compare the difference of survival rate in each group. Results: A total of 234 repairs for maxillary defects were performed in 219 patients. Fibula flaps were used in 4/13 of class Ⅰ defects; temporal muscle flaps (11/24, 45.8%) and anterolateral thigh flaps (6/24, 25.0%) used in class Ⅱ defects; temporal muscle flaps (71/128, 55.5%), anterolateral thigh flaps (6/24, 25.0%) and fibula flaps (12/128, 9.4%) used in class Ⅲ defects; and anterolateral thigh flaps (8/20, 40.0%) and rectus abdominis flaps (8/20, 40.0%) used in class Ⅳ defects. The success rate of local pedicled flaps was 95.6% (109/114) and that of free flaps was 95.8% (115/120). Thrombosis(10/234,4.3%) was a main reason for repair failure. Among the followed-up 88 patients, swallowing and speech functions recovered, 82 (93.2%) of them were satisfied with appearance, and 75 (85.2%) were satisfied with visual field. The 3-year and 5-year overall survival rates were 66.5% and 63.6%, and the 3-year and 5-year disease-free survival rates were 57.1% and 46.2%, respectively, in the 169 patients with malignant tumors. Conclusion: A new classification of maxillary defects is proposed, on which suitable flaps are selected to offer patients good functional and aesthetic outcomes and high quality of life.

Meta-analysis of the effect of different concentrations of atropine inhibiting spherical equivalent degree and axial length of myopia in children / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To systematically evaluate the effects of 0.01%, 0.025% and 0.05% ophthalmic atropine on the change of spherical equivalent(SE)degree and axial length(AL)of myopic children. METHODS: PubMed, Embase, Cochrane Library, Web of Science, CNKI, Wanfang Database, VIP and CBM were searched for all publications related to 0.01%, 0.025% and 0.05% atropine to control myopia simultaneously. The publication time is from the database construction to May 2022. The Cochrane handbook was used to evaluate the risk of bias and quality of the included literature, STATA12.0 was used to detect publication bias and Revman5.4 software was used for Meta-analysis. RESULTS: A total of 6 literatures(1 239 eyes)were included, with 5 randomized controlled trials and 1 case-control study. Meta-analysis results showed that 0.025% atropine had better inhibitory effect on SE and AL than 0.01% atropine(SE: WMD=-0.15, 95%CI: -0.23--0.06, P&#x003C;0.001; AL: WMD=0.07, 95%CI: 0.03-0.10, P&#x003C;0.001). The inhibitory effect of 0.05% atropine on SE and AL was better than 0.01% atropine(SE: WMD=-0.35, 95%CI: -0.44--0.26, P&#x003C;0.001; AL: WMD=0.16, 95%CI: 0.12-0.20, P&#x003C;0.001). The inhibitory effect of 0.05% atropine on SE and AL increase was better than 0.025% atropine(SE: WMD=-0.20, 95%CI: -0.28--0.11, P&#x003C;0.001; AL: WMD=0.09, 95%CI: 0.06-0.12, P&#x003C;0.001). CONCLUSION: The concentration of 0.05% atropine is superior to 0.01% and 0.025% atropine in the control of SE and AL. However, the side effects of long-term use remain to be observed.

Reference gene screening for Real-time quantitative PCR in Polygonum multiflorum / 中国中药杂志

To select suitable references gene of Polygonum multiflorum for gene expression analysis in different tissues, five candidate reference genes like Actin,GAPDH,SAND,PP2A,TIP41 were selected from the transcriptome data of P. multiflorum, then the specific primers were designed. The expression stability of the five reference genes in different tissues of P. multiflorum was analyzed by Real-time quantitative PCR through avilable analysis methods such as geNorm, NormFinder, BestKeeper, Delta CT and RefFinder, to ensure the reliability of the analysis results. The results showed that there were significant differences in the expression levels and stability of candidate genes in different tissues of P. multiflorum. Ct distribution analysis of the expression levels of candidate genes showed that the expression levels of Actin and GAPDH genes were relatively high in different tissues, while the expression levels of SAND, PP2A and TIP41 were lower. The stability of each candidate gene was analyzed by different methods. The results of geNorm analysis showed that the expression of PP2A and GAPDH was the most stable, the expression stability of SAND was the worst, the stability of PP2A was the highest in both NormFinder and Delta CT, the stability of SAND was the lowest, and the stability of Actin was the most stable in BestKeeper analysis. Through the comprehensive evaluation and analysis of the stability of candidate genes by RefFinder, it is concluded that the stability of PP2A gene is the highest, followed by GAPDH, Actin, TIP41, SAND, and SAND gene is the worst. Therefore, the PP2A gene is an ideal reference gene for the analysis of gene expression in different tissues of P. multiflorum.

Role of P2X 7 receptor in microglia in medial prefrontal cortex in neuropathic pain in rats: relationship with autophagy / 中华麻醉学杂志

Objective:To evaluate the role of P2X 7 receptor in microglia in the medial prefrontal cortex (mPFC) in neuropathic pain (NP) and the relationship with autophagy in rats. Methods:Sixty-four healthy SPF male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-250 g, were divided into 4 groups ( n=16 each) using a random number table method: sham operation group (S group), NP group, sham operation+ P2X 7 receptor blocking group (SP group), and NP+ P2X 7 receptor blocking group (NP+ P group). The NP model was established by ligation of the sciatic nerve.Fourteen days later a cannula was placed in the mPFC with a brain stereotactic instrument, P2X 7 receptor blocker A-740003 0.5 μg/0.5 μl was injected into bilateral mPFC for 3 consecutive days starting from the 14th day in SP and NP+ P groups, and DMSO 0.5 μl was injected instead of A-740003 in S and NP groups.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 3, 7 and 10 days after establishing the model and 14, 15 and 16 days after administration.Then the rats were sacrificed, and the mPFC was removed for determination of the expression of P2X 7 receptor and mRNA and autophagy-related proteins Beclin1 and LC3Ⅱ/Ⅰ (by quantitative real-time polymerase chain reaction or by Western blot) and co-expression of P2X 7R and microglia (by immunofluorescence) and the number of autophagosomes in mPFC (with a transmission electron microscope). Results:Compared with group S, MWT was significantly decreased, and TWL was shortened at 3, 7 and 10 days after establishing the model, the expression of P2X 7 receptor and mRNA, Beclin1 and LC3Ⅱ/Ⅰ was up-regulated at 30 min after administration on 16 days after establishing the model, and the number of cells co-expressing P2X 7 receptor and IBA-1 and the number of autophagosomes were increased in NP and NP+ P groups ( P<0.05), and no significant change was found in the indexes mentioned above in group SP ( P>0.05). Compared with group NP, MWT was significantly increased, and TWL was prolonged at 30 min after administration on 14, 15 and 16 days after establishing the model, the expression of P2X 7 receptor and mRNA, Beclin1 and LC3Ⅱ/Ⅰ was down-regulated, and the number of cells co-expressing P2X 7 receptor and Iba-1 and the number of autophagosome were decreased in group NP+ P ( P<0.05). Conclusion:Up-regulation of P2X 7 receptor expression in microglia in mPFC is involved in the process of NP in rats, which is associated with the promotion of autophagy.

Screening of plant growth-promoting rhizobacteria and its effect on seed germination of Polygonum multiflorum / 中国中药杂志

In this study, the rhizobacteria and actinomycetes of Polygonum multiflorum were screened for the strains with indole acetic acid(IAA)-producing capacity by Salkowski method, the siderophore-producing strains by Chrome Azurol S(CAS) assay, and the strains with inorganic phosphorus-solubilizing capacity by PKO inorganic phosphorus medium. The strains were identified by morphological identification, physiological and biochemical characteristics, and 16 S rDNA sequences. Furthermore, the effect of growth-promoting strains on the seed germination and development of P. multiflorum was tested. The results showed that among 196 strains, two strains F17 and F42 were found to be capable of producing IAA and siderophore and solubilizing inorganic phosphorus simulta-neously. For F17 and F42, the results are listed below: 38.65 and 33.64 mg·L~(-1) for IAA production, 0.85 and 0.49 for siderophore-producing capacities(A_s/A_r), and 1.35 and 1.70 for inorganic phosphorus-solubilizing capacities(D/d), respectively. Comprehensive analysis revealed that strains F17 and F42 were identified as Pseudochrobactrum asacharolyticum and Bacillus aryabhattai, respectively, and both could significantly promote the seed germination of P. multiflorum.

Clinical comparative study of free posterior tibial artery perforator flap and radial forearm free flap for head and neck reconstruction / 中华耳鼻咽喉头颈外科杂志

Objective: To compare the clinical application results of the FPTF (free posterior tibial artery perforator flap) and RFFF (radial forearm free flap) for reconstruction of head and neck defects. Methods: A retrospective analysis of 27 cases treated with FPTF (19 males and 8 females, aged 14-69 years) and 24 cases with RFFF (11 males and 13 females, aged 22-69 years) for head and neck defect reconstruction at Beijing Tongren Hospital of Capital Medical University from January 2015 to December 2020 was conducted. Flap size, vascular pedicle length, matching degree of recipient area blood vessels, preparation time, total operation time, hospital stay, recipient area complications, donor area complications and scale-based patient satisfaction were compared between two groups of patients with FTPF and RFFF. SPSS 26.0 statistical software was used for statistical analysis. Results: There was no statistically significant difference between the two groups of patients in tumor T staging (P=0.38), primary sites (P=0.05) and mean flap areas ((53.67±29.84) cm2 vs. (41.13±11.08) cm2, t=-1.472, P=0.14). However the mean vascular pedicle length of FPTF was more than that of RFFF ((11.15±2.48)cm vs. (8.50±1.69)cm, t=-4.071, P<0.01). The donor sites of 4 patients in FPTF group could be sutured directly, while all the 24 patients in RFFF group received skin grafts from the donor sites. There was no statistically significant difference in the recipient area arteries between two groups of flaps (P=0.10), with more commonly using of the facial artery (RFFF: FPTF=21∶27), but there was significant difference in the recipient area veins (P<0.01), with more commonly using of the external jugular vein in RFFF (14/24) than FPTF (4/32) and the posterior facial vein in FPTF (27/32) than RFFF (9/24). There were 10 recipient complications and 3 donor complications in RFFF group; no recipient complication and 3 donor complications occurred in FPTF group. With patient's subjective evaluation of the donor site at 12 months after surgery, FPTF was better than RFFF (χ²=22.241, P<0.01). Conclusions: FPTF is an alternative to RFFF in head and neck reconstruction and has unique advantages in aesthetics and clinical application.

Retrospective study of thyroid invasion and central lymph node metastasis in 124 patients with hypopharyngeal carcinoma / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the characteristics of thyroid invasion and central lymph node metastasis of hypopharyngeal carcinoma, and the impact on survival rate and quality of life. Methods: A retrospective analysis of 124 cases (122 males and 2 females with age range from 36 to 78 years old) with laryngopharyngeal squamous cell carcinoma who were initially treated in the Department of Head and Neck Surgery, Beijing Tongren Hospital Affiliated to Capital Medical University from January 2014 to December 2017 was performed. The clinical data included tumor location, pathological T stage, pathological N stage, invasion of thyroid gland, central lymph node metastasis, surgical procedures and so on. Patients were grouped according to if presence of thyroid invasion and central lymph node metastasis. With follow-up, the survival was analyzed by Kaplan-Meier method, and tumor recurrence and metastasis were evaluated. Results: Of the patients, 12 patients had thyroid involvement and 5 patients had central lymph node metastasis. The incidence of thyroid involvement was 8.16% (8/98) in pyriform sinus, 1/18 in posterior pharyngeal wall and 3/8 in posterior cricoid wall, with statistically significant difference (χ2=15.076,P=0.008). The incidence of central lymph node metastasis was 1.02% (1/98) in pyriform sinus, 3/18 in posterior pharyngeal wall and 1/8 in posterior cricoid wall, also with statistically significant difference (χ2=11.205, P=0.008). There was no statistical correlation between thyroid invasion or central lymph node metastasis and gender, smoking or alcohol exposure history and tumor pathological differentiation (all P>0.05). The 3-year overall survival rate was 80.65% and the 3-year recurrence free rate was 85.48%. Totally 24 patients died in 3 years, including 4 cases in thyroid invasion group and 1 case in central lymph node metastasis group. Local recurrence occurred in 18 patients, including 4 cases in thyroid invasion group and 1 case in central lymph node metastasis group. There was no significant difference in survival between patients with and without thyroid invasion and central lymph node metastasis (all P>0.05). There were significantly difference in 3-year overall survival and relapse-free survival among the groups with different T stages, N stages, pathological stages and tumor pathological differentiation levels (all P<0.05). There were significantly differences in the levels of serum calcium and FT3 between the groups with or without thyroid invasion and central lymph node metastasis (all P<0.05). Conclusion: The incidences of thyroid invasion and central lymph node metastasis of hypopharyngeal carcinoma are rare, and the risk of occurrence is related to the primary site of tumor. Comprehensive evaluation, correct decision-making and accurate treatment could be helpful to cure radically the tumor, to prevent recurrence and to improve the quality of life of patients.

Application of deep convolutional neural networks in the diagnosis of laryngeal squamous cell carcinoma based on narrow band imaging endoscopy / 中华耳鼻咽喉头颈外科杂志

Objective: To explore the possibility of using artificial intelligence (AI) technology based on convolutional neural network (CNN) to assist the clinical diagnosis of laryngeal squamous cell carcinoma (LSCC) through deep learning algorithm. Methods: A deep CNN was developed and applied in narrow band imaging (NBI) endoscopy of 4 799 patients with laryngeal lesions, including 3 168 males and 1 631 females, aged from 21 to 87 years, from 2015 to 2017 in Beijing Tongren Hospital, Capital Medical University. A simple randomization method was used to select the laryngeal NBI images of 2 427 patients (1 388 benign lesions and 1 039 LSCC lesions) for the training and correction the CNN model. The remaining laryngeal NBI images of 2 372 patients (including 1 276 benign lesions and 1 096 LSCC lesions) were used as validation data set to compare performance between CNN and otolaryngologists. SPSS 21.0 software was used for Chi-square test to calculate the accuracy, sensitivity and specificity of AI and otolaryngologists. The area under the curve (AUC) of receiver operating curve (ROC) was used to evaluate the diagnostic ability of the algorithm for NBI images. Results: The accuracy, sensitivity and specificity for NBI predictions were respectively 90.91% (AUC=0.96), 90.12% and 91.53%, which were equivalent to those for otolaryngologists' predictions (accuracy, sensitivity and specificity were (91.93±3.20)%, (91.33±3.25)% and (93.02±2.59)%, t values were 0.64, 0.75 and 1.17, and P values were 0.32, 0.28 and 0.21, respectively). The diagnostic efficiency of CNN was significantly higher than that of otolaryngologists (0.01 vs. 5.50, t =9.15, P<0.001). Conclusion: AI based on deep CNN is effective for using in the laryngeal NBI image diagnosis, showing a good application prospect in the diagnosis of LSCC.

RIP1-dependent linear and nonlinear recruitments of caspase-8 and RIP3 respectively to necrosome specify distinct cell death outcomes

There remains a significant gap in our quantitative understanding of crosstalk between apoptosis and necroptosis pathways. By employing the SWATH-MS technique, we quantified absolute amounts of up to thousands of proteins in dynamic assembling/de-assembling of TNF signaling complexes. Combining SWATH-MS-based network modeling and experimental validation, we found that when RIP1 level is below ~1000 molecules/cell (mpc), the cell solely undergoes TRADD-dependent apoptosis. When RIP1 is above ~1000 mpc, pro-caspase-8 and RIP3 are recruited to necrosome respectively with linear and nonlinear dependence on RIP1 amount, which well explains the co-occurrence of apoptosis and necroptosis and the paradoxical observations that RIP1 is required for necroptosis but its increase down-regulates necroptosis. Higher amount of RIP1 (>~46,000 mpc) suppresses apoptosis, leading to necroptosis alone. The relation between RIP1 level and occurrence of necroptosis or total cell death is biphasic. Our study provides a resource for encoding the complexity of TNF signaling and a quantitative picture how distinct dynamic interplay among proteins function as basis sets in signaling complexes, enabling RIP1 to play diverse roles in governing cell fate decisions.

Effect of Polygonum multiflorum-Andrographis paniculata intercropping system on rhizosphere soil actinomycetes community structure and diversity of P. multiflorum / 中国中药杂志

To investigate the effect of Polygonum multiflorum-Andrographis paniculata intercropping system on rhizosphere soil actinomycetes of P. multiflorum, the community structure and diversity of soil actinomycetes were studied by using the original soil as the control group and the rhizosphere soil actinomycetes communities of P. multiflorum under monoculture and intercropping systems as the experimental group. In this study 655 221 effective sequences were obtained with an average length of 408 bp. OTU coverage and rarefaction curve showed that the sequencing could represent the real situation of soil actinomycetes. According to the results of alpha diversity analysis, the diversity soil actinomycetes varied as follows: original soil>intercropping soil>monoculture soil. The soil actinomycetes community structure and the relative abundance of dominant genera were significantly changed by both monoculture and intercropping, especially monoculture. OTU clustering and PCA analysis of soil samples showed that all the soil samples were divided into three distinct groups and the original soil was more similar to intercropping soil. In addition, intercropping increased the relative abundance of some beneficial actinomyces, such as Kitasatospora and Mycobacterium, which was beneficial to maintain soil health and reduce the occurrence of soil-borne diseases. The results show that, P. multiflorum-A. paniculata intercropping reduced the change of community structure and the decrease of diversity of soil actinomycetes caused by P. multiflorum monoculture, and made the actinomycete community in rhizosphere soil of P. multiflorum close to the original soil.

Relationship between P2X 7 receptors and NLRP3/IL-1β pathway in spinal neurons in development of inflammatory pain in rats / 中华麻醉学杂志

Objective:To evaluate the relationship between P2X 7 receptors and nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3)/interleukin-1beta (IL-1β) pathway in spinal neurons in the development of inflammatory pain (IP) in rats. Methods:SPF healthy adult male Wistar rats, weighing 180-220 g, were used in this study.Forty rats in which intrathecal catheters were successfully implanted were divided into 5 groups ( n=8 each) using a random number table method: control group (group CON), group IP, IP plus dimethyl sulfoxide (DMSO) group (group IP-DMSO), IP plus P2X 7 receptor antagonist A740003 group (group IP-A) and IP plus P2X 7 receptor agonist ATP group (group IP-ATP). Rats were anesthetized with pentobarbital sodium 40 mg/kg.IP was induced by injecting complete Freund′s adjuvant 50 μl into the right ankle joint cavity, while group CON was injected with the equal volume of normal saline instead.On 1 day before establishing the model, immediately after establishing the model, and on 1, 2 and 3 days after establishing the model, 1% DMSO 10 μl was intrathecally injected once a day in group IP-DMSO, A740003 0.1 nmol(dissolved in DMSO 10 μl) was intrathecally injected once a day in group IP-A, and ATP 150 nmol(dissolved in DMSO 10 μl) was injected intrathecally once a day in group IP-ATP.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured on 3 days after establishing the model.Enzyme-linked immunosorbent assay was used to determine the prostaglandin E2 (PGE2) concentrations in right ankle tissues and IL-1β concentrations in cerebrospinal fluid (CSF). Then rats were sacrificed, and the lumber segments (L 4-6) of the spinal cord were removed for determination of the expression of NLRP3, casepase-1, IL-1β (by Western blot) and co-expression of P2X 7 receptors with neuron-specific nucleoprotein (NeuN) and NLRP3 and with NeuN (by immunofluorescence). Results:Compared with group CON, PGE2 contents in ankle tissues were significantly increased in group IP, and the MWT was significantly decreased, the TWL was shortened, the concentrations of IL-1β in CSF were increased, and the expression of NLRP3, caspase-1 and IL-1β was up-regulated in the other four groups ( P<0.05). Compared with group IP, the MWT was significantly increased, the TWL was prolonged, the concentrations of IL-1β in CSF were decreased, and the expression of NLRP3, caspase-1 and IL-1β was down-regulated in group IP-A ( P<0.05), the MWT was significantly decreased, TWL was shortened, the concentrations of IL-1β in CSF were increased, and the expression of NLRP3, caspase-1 and IL-1β was up-regulated in group IP-ATP ( P<0.05), and no significant change was found in the parameters mentioned above in group IP-DMSO ( P>0.05). P2X 7 was co-expressed with NeuN, and NLRP3 was co-expressed with NeuN. Conclusion:P2X 7 receptors in spinal neurons are involved in the development of inflammatory pain by activating NLRP3/IL-1β signaling pathway in rats.

Relationship between histone acetylation and NR2B-CREB-BDNF signaling pathway during sevoflurane-induced cognitive decline in aged mice / 中华麻醉学杂志

Objective:To evaluate the relationship between histone acetylation and N-methyl-D-aspartic acid (NMDA) receptor subunit 2B (NR2B)-cAMP-response element binding protein (CREB)-brain-derived neurotrophic factor (BDNF) signaling pathway during sevoflurane-induced cognitive decline in aged mice.Methods:Forty-eight SPF healthy male C57BL/6J mice, aged 22 months, weighing 32-40 g, were divided into 4 groups ( n = 12 each) using a random number table method: control group (group C), sevoflurane group (group S), dimethyl sulfoxide (DMSO) plus sevoflurane group (group DS) and histone deacetylase inhibitor vorinostat (SAHA) plus sevoflurane group (group SS). The 100% oxygen was inhaled for 2 h in group C. In S, DS and SS groups, 3.0% sevoflurane was inhaled for 2 h, Morris Water Maze (MWM) test was performed at 24 h after the end of sevoflurane inhalation.SAHA 50 mg/kg was intraperitoneally injected in group SS, and the equal volume of DMSO was given instead in group DS at 2 h before sevoflurane inhalation and at 2 h before MWM test was performed every day.Animals were sacrificed after the MWM test, and hippocampi were isolated for determination of the expression of acetyl-H3 in the nucleus and NR2B, phosphorylated CREB (p-CREB) and BDNF in cytoplasm by Western blot.Real-time polymerase chain reaction was used to determine the expression of NR2B and BDNF mRNA. Results:Compared with group C, the escape latency was significantly prolonged, and the percentage of time spent in target quadrant was decreased, and the expression of acetyl-H3, NR2B, p-CREB, BDNF, NR2B mRNA, and BDNF mRNA was down-regulated in S, DS and SS groups ( P<0.05). Compared with group S or group DS, the escape latency was significantly shortened, the percentage of time spent in target quadrant was increased, the expression of acetyl-H3, NR2B, p-CREB, BDNF, NR2B mRNA, and BDNF mRNA was up-regulated in group SS ( P<0.05). There was no significant difference in each parameter mentioned above between group S and group SD ( P>0.05). Conclusion:Histone acetylation is involved in the pathophysiological mechanism of cognitive decline induced by sevoflurane anesthesia by regulating NR2B-CREB-BDNF signaling pathway in aged mice.

Analysis for potential targeting genes of TPF regimen induction chemotherapy in hypopharyngeal squamous cell carcinoma / 中华耳鼻咽喉头颈外科杂志

Objective@#To analyze the differentially expressed genes related to the chemosensitivity with the TPF regimen for hypopharyngeal squamous cell carcinoma and to measure potential functional targeting genes expressions.@*Methods@#Twenty-nine patients with primary hypopharyngeal cancer who underwent induction chemotherapy with TPF from January 2013 to December 2017 in Beijing Tongren Hospital were enrolled for microarray analysis, including 28 males and 1 female, aged from 43 to 73 years old. Among them, 16 patients were sensitive to chemotherapy while 13 patients were non-sensitive. Illumina Human HT-12 Bead Chip was applied to analyze the gene expressions and online bioinformatics analysis was used to analyze the differentially expressed genes. Reverse transcription and quantitative real-time PCR (RT-qPCR) was used to measure the mRNA expression of potential functional genes of TPF induction chemotherapy in 43 samples, 29 from original patients and 14 from additional patients. Graphpad prism 7.0 software was used for statistical analysis.@*Results@#A total of 1 381 significantly differentially expressed genes were screened out. By GO analysis, up-regulated genes included sequestering in extracellular matrix, chemokine receptor binding and potassium channel regulator activity; down-regulated genes included regulation of angiogenesis, calcium ion binding and natural killer cell activation involved in immune response. With KEGG database analysis, down-regulated pathways included ECM-receptor interaction and peroxisome and up-regulated pathways included Glutathione metabolism and PPAR signaling pathway. The expressions of CD44 and IL-6R were significantly different and appeared biologically significant. CD44 was significantly upregulated in insensitive tissues (0.54±0.06) compared with sensitive tissues (0.33±0.04)(P<0.01). IL-6R was significantly downregulated in insensitive tissues (0.44±0.03) compared with sensitive tissues. (0.68±0.03) (P<0.01).@*Conclusion@#CD44 and IL-6R may be potentially functional genes of TPF induction chemotherapy in hypopharyngeal squamous cell carcinoma.