Genetic analysis of a Chinese pedigree with 18q21.2-q22.3 duplication and deletion in two offspring respectively resulting from a maternal intrachromosomal insertion / 中华医学遗传学杂志

OBJECTIVE@#To provide prenatal diagnosis, pedigree analysis and genetic counseling for a pregnant woman who had given birth to a child featuring global developmental delay.@*METHODS@#A pregnant woman who underwent prenatal diagnosis at the Affiliated Hospital of Southwest Medical University in August 2021 was selected as the study subject. Peripheral blood samples were collected from the woman, her husband and child, in addition with amniotic fluid sample during mid-pregnancy. Genetic variants were detected by G-banded karyotyping analysis and copy number variation sequencing (CNV-seq). Pathogenicity of the variant was predicted based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). Candidate variant was traced in the pedigree to assess the recurrence risk.@*RESULTS@#The karyotypes of the pregnant woman, her fetus, and affected child were 46,XX,ins(18)(p11.2q21q22), 46,X?,rec(18)dup(18)(q21q22)ins(18)(p11.2q21q22)mat and 46,XY,rec(18)del(18)(q21q22)ins(18)(p11.2q21q22)mat, respectively. Her husband was found to have a normal karyotype. CNV-seq has revealed a 19.73 Mb duplication at 18q21.2-q22.3 in the fetus and a 19.77 Mb deletion at 18q21.2-q22.3 in her child. The duplication and deletion fragments were identical to the insertional fragment in the pregnant woman. Based on the ACMG guidelines, the duplication and deletion fragments were both predicted to be pathogenic.@*CONCLUSION@#The intrachromosomal insertion of 18q21.2-q22.3 carried by the pregnant woman had probably given rise to the 18q21.2-q22.3 duplication and deletion in the two offspring. Above finding has provided a basis for genetic counseling for this pedigree.

SENP1 induced protein deSUMO modification increased the chemotherapy sensitivity of endometrial cancer side population cells / 中华肿瘤杂志

Objective: To inhibit the stemness maintenance potential of endometrial cancer and increase the sensitivity of endometrial cancer side population cells to chemotherapy drugs by inducing extensive deSUMOylation modification of proteins. Methods: Flow cytometry was used to sort and culture CD133(+) CD44(+) KLE endometrial cancer cell clone spheres. Protein expression level of small ubiquitin-related modifier 1 (SUMO1) and two stemness maintenance genes of tumor side population cells, octamer binding transcription factor-4 (Oct4) and sex determining region Y-box2 (Sox2), were detected by western blotting method. Lentivirus-mediated Sentrin/SUMO-specific proteases 1 (SENP1) gene was stably transfected into KLE side population cells. Western blotting was used to detect the protein expressions of SENP1, SUMO1, Oct4 and Sox2. The clone formation rate was compared between KLE side population cells with or without SENP1 overexpression. Flow cytometry was applied to detect cell cycle changes. 3-(4, 5-Dimethylthiazole-2)-2, 5-diphenyl-tetrazolium bromide (MTT) experiment and flow cytometry apoptosis method were used to detect the chemosensitivity of the side population of endometrial cancer cells to cisplatin. Tumor-bearing mouse models of endometrial cancer were established to detect the effect of SENP1 overexpression on the chemotherapy sensitivity of cisplatin. Results: Compared with CD133(-)CD44(-) KLE cells, CD133(+) CD44(+) KLE side population cells could form clonal spheres and express higher levels of SUMO1, Oct4 and Sox2 proteins (P<0.05). Compared with KLE side population cells that were not transfected with SENP1 gene, the expression level of SENP1 protein in KLE side population cells overexpressing SUMO1、Oct4 and Sox2 were lower. The clonal sphere formation rate was reduced from (25.67±5.44)% to (7.46±1.42)%, and cell cycle shifted from G(0)/G(1) phase to G(2) phase. IC(50) of cisplatin decreased from (55.46±6.14) μg/ml to (11.55±3.12) μg/ml, and cell apoptosis rate increased from (9.76±2.09)% to (16.79±3.44)%. Overexpression of SENP1 could reduce the tumorigenesis rate of KLE side population cells in vivo and increase their chemotherapy sensitivity to cisplatin (P<0.05). Conclusion: Overexpression of SENP1 can induce protein deSUMOylation modification, inhibit the stemness maintenance potential of endometrial cancer side population cells, and enhance their chemotherapy sensitivity, which provides a new reference for gene therapy of endometrial cancer.

Desmethylbellidifolin protects against chronic alcoholic fatty liver disease by regulating Akt-mTORC1 pathway mediated autophagy / 中国药理学与毒理学杂志

OBJECTIVE To investigate the protective effect and potential mechanism of desmethylbellidifolin (DMB) in chronic alcoholic fatty liver disease. METHODS C57BL/6 mice were randomly divided into five groups. Control, meta?doxine and DMB group (high dose and low dose) mice were fed with Lieber-DeCarli liquid diet containing 5％alcohol for six weeks. Pair-fed group mice were fed with a liquid diet containing the same calories. After treatment, serum GOT, GPT, TG and hepatic T-CHO, TG, GSH, GSH-Px, SOD and CAT levels were measured. Ectopic liver lipid deposition was determined by oil red O and hematoxylin-eosin (HE) staining. Lipid metabolism and autophagy related genes expression were determined by real-time PCR and Western blotting. Electron microscope and laser scanning confocal microscope were used to detect autophagosome and autophagy flux. RESULTS DMB treatment markedly reduced serum TG, GOT and GPT levels in alcohol-induced mice, as well as hepatic levels of T-CHO, TG and MDA, while increased the GSH, GSH-Px, SOD and CAT levels in the liver. Oil red O and HE staining showed that the alcohol-induced lipid accumulation and hepatocyte morphology changes were significantly improved by DMB treatment. Mecha?nistically, the expression of stearoyl-CoA desaturase 1 and fatty acid synthase were significantly decreased, while lipoly?sis related hormone-sensitive lipase was elevated in mouse liver by DMB treatment. In addition, DMB could inhibit the phosphorylation of Akt and mTORC1, and activate autophagy process by inducing autophagy related genes expression, such as LC3, ATG5 and ATG7. Moreover, treatment with DMB notably increased the number of autolysosome and promote the autophagy flux, which may therefore induce the lipolysis and oxidation of lipids and prevent the alcohol-induced excessive lipid accumulation in the liver. CONCLUSION DMB exerts a protective role in alcoholic fatty liver dis?ease by regulating the Akt-mTORC1 pathway mediated autophagy activation.

Feasibility and significance of early vitrectomy for open ocular trauma / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To explore the feasibility and significance of early vitrectomy for open ocular trauma.<p>METHODS: Eighty-two cases(82 eyes)of open ocular trauma complicated with retinal detachment in Department of Ophthalmic Surgery in our hospital from March, 2018 to 2019 were reviewed, and the patients were divided into early group(<7d)and routine group(7-14d)according to the time of vitrectomy. All patients were followed up for 3mo. The overall efficacy, visual acuity changes, retinal detachment reduction rate, dysfunctional vitreous diseases(TPVR)and complications were compared between the two groups.<p>RESULTS: The total cure rate and visual acuity improvement rate of the early group were 93% and 95%, and those of the conventional group were 71% and 76%. The early group was significantly higher than the conventional group, and the difference was statistically significant(χ2=5.222, 4.783, <i>P</i><0.05). The complication rates of silicone oil dependence, vitreous hemorrhage and secondary glaucoma were 5%, 10% and 5% in the early group which were significantly lower than in the conventional group(27%, 39%, 29%, <i>P</i><0.05). There was no difference in the reduction rate of retinal detachment between two groups(<i>P</i>>0.05). The incidence of TPVR was 5% in the early group, which lower than in the conventional group,but had no difference(<i>P</i>>0.05).<p>CONCLUSION: For patients with open ocular trauma and retinal detachment, early vitrectomy may be more effective, which is more significant in improving the rate of retinal reduction and reducing the incidence of TPVR.

Improved synthesis process of optical-magnetic bimodal probe of Gd-4, 7-Bis-carboxymethyl-10-(2-fluoresceinthioureaethyl)-1, 4, 7, 10-tetraaza-cyclododec-1-yl-acetic acid complexes / 北京大学学报(医学版)

OBJECTIVE@#To improve the methods to synthesize and purify of optical-magnetic bimodal molecular probe of Gd-[4, 7-Bis-carboxymethyl-10-(2-fluorescein thioureaethyl)-1, 4, 7, 10-tetraaza-cyclododec-1-yl]-acetic acid complexes.@*METHODS@#Target compound (7), optical-magnetic bimodal molecular molecular probe, was synthesized by the use of 1, 4, 7, 10-tetraazacyclododecane (1) as starting material via substitution reaction, hydrolysis reaction, coupling reaction and complexation reaction with metal.@*RESULTS@#The synthetic route of Gd-[4, 7-Bis-carboxymethyl-10-(2-fluoresceinthioureaethyl)-1, 4, 7, 10-tetraaza-cyclododec-1-yl]-acetic acid complexes was improved. The optical-magnetic bimodal molecular probes were synthesized by substitution reaction, hydrolysis reaction, coupling reaction and complex reaction with metal respectively. For the improved route, the total yield could reach 34.6% which was higher than the original route (18.0%). The structures of those compounds were identified by 1H nuclear magnetic resonance, 13C nuclear magnetic resonance, and mass spectrometry. The improved route could avoid the uncontrollable disadvantage of the substitution reaction, this process could reduce the formation of impurities and made the purification process easier, and in the aspect of purification and separation, the preparative high-performance liquid chromatography with less sample loading and high cost was improved to a column chromatography with many sample loads and being easy to operate. Therefore, the use of column chromatography could be more conducive to mass production of the optical-magnetic bimodal molecular molecular probe.@*CONCLUSION@#The improved synthetic route improves the controllability of the reaction conditions and makes it easier to purify and separate the compounds. At the same time, the improved synthetic route can increase the total yield significantly. The optical-magnetic bimodal molecular probe can combine the living magnetic resonance imaging with the in vitro optical imaging to realize the dual synchronous detection of magneto-optics, so that the detection results of the living magnetic resonance imaging and the in vitro optical imaging are mutually verified. In other words, this synthetic optical-magnetic bimodal molecular probe will make the experimental results more accurate and reliable. In subsequent biological experimental studies, the optical-magnetic bimodal molecular probe can be applied to related research of brain structure and function, and the probe can be used for the brain-related diseases researches, such as brain tumors. after intravenous administration, and thus the optical-magnetic bimodal molecular probe can play an important role in medical treatment of brain tumors and cerebrovascular diseases.

Influence of Dexmedetomidine Applied in Combined Acupuncture-medication Anesthesia on Vomiting After Thyroidectomy / 上海针灸杂志

Objective To observe the influence of dexmedetomidine applied in combined acupuncture-medication anesthesia on vomiting after thyroidectomy. Method Seventy patients who were going to receive thyroidectomy were randomized into group A and B, with 35 cases in each group. Combined acupuncture-medication anesthesia [electroacupuncture at Hegu (LI4), Neiguan (PC6) and Futu (ST32) plus intravenous infusions of sufentanil citrate injection (0.3 mg/kg) 20 minutes before operation]was adopted in both groups. Group A was additionally intervened by constantvelocity micropump infusion of dexmedetomidine hydrochloride injection (0.5 mg/kg) within 10 minutes before operation and remaining 0.4 mg/kg/min during operation, while group B was intervened by infusion of normal saline 10 minutes before operation. The observer's assessment of awareness/sedation (OAA/S) scores at different time points[lying in bed for 5 minutes before operation (T0), before the beginning of the operation (T1), 30 minutes after operation (T2), 60 minutes after operation (T3) and at the end of operation (T4)] in the two groups were observed. The additional times and total dose of sufentanil during operation, the number of cases using esmolol and urapidil, and the number of vomiting cases occurred within 2 hours after operation and 2～24 hours after operation in the two groups were recorded.Result The additional times and total dose of sufentanil during operation in group A were significantly different from those in group B (P<0.01). The utilization rate of esmolol and urapidil were respectively 31.4％ and 14.3％ in group A versus 77.1％ and 65.7％ in group B, and the between-group differences were statistically significant (P<0.01). The incidence of vomiting within 2 hours after operation and 2～24 hours after operation were respectively 20.0％ and 17.1％in group A versus 54.3％ and 42.9％ in group B, and the between-group differences were statistically significant (P<0.01). The OAA/S scores at different time points (T1, T2, T3 and T4) in group A were significantly different from those in group B (P<0.01). Conclusion Dexmedetomidine applied in combined acupuncture-medication anesthesia can reduce the additional times and total dose of sufentanil during thyroidectomy, and it can reduce the incidence of postoperative vomiting as well.

Effect of loureirin A against Candida albicans biofilms / 中国天然药物

Loureirin A is a major active component of Draconis sanguis, a traditional Chinese medicine. This work aimed to investigate the activity of loureirin A against Candida albicans biofilms. 2, 3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT)reduction assay and scanning electron microscopy were used to investigate the anti-biofilm effect. Minimal inhibitory concentration testing and time-kill curve assay were used to evaluate fungicidal activity. Cell surface hydrophobicity (CSH) assay and hyphal formation experiment were respectively carried out to investigate adhesion and morphological transition, two virulence traits of C. albicans. Real-time RT-PCR was used to investigate gene expression. Galleria mellonella-C. albicans and Caenorhabditis elegans-C. albicans infection models were used to evaluate the in-vivo antifungal effect. Human umbilical vein endothelial cells and C. elegans nematodes were used to evaluate the toxicity ofloureirin A. Our data indicated that loureirin A had a significant effect on inhibiting C. albicans biofilms, decreasing CSH, and suppressing hyphal formation. Consistently, loureirin A down-regulated the expression of some adhesion-related genes and hypha/biofilm-related genes. Moreover, loureirin A prolonged the survival of Galleria mellonella and Caenorhabditis elegans in C. albicans infection models and exhibited low toxicity. Collectively, loureirin A inhibits fungal biofilms, and this effect may be associated with the suppression of pathogenic traits, adhesion and hyphal formation.

Effects of the Survivin shRNA-APC double-gene on the expressions of survivin,Cytc,Smac and Caspase-9 in the colon cancer mitochondrial apoptosis pathway / 医学研究生学报

Objective Little is known about the effect of RNAi on mitochondrial apoptotic pathways. This study aims to explore the effects of the Survivin shRNA-APC double-gene on colon cancer mitochondrial apoptosis pathway-related factors survivin,cytochrome C (Cytc),second mitochondria-derived activator of caspases (Smac),and cysteine aspartic acid specific protease 9 (Caspase-9) as well as on the apoptosis of colon cancer transplanted tumor (CCTT) cells. Methods Thirty nude mice were randomly divided into five groups of equal number,Survivin shRNA-APC double-gene,survivin shRNA,APC,empty vector and blank transfection. The CCTT model was established in the nude mice by subcutaneous injection of the colon cancer cell strains stably transfected with the Survivin shRNA-APC double-gene,survivin shRNA,APC,an empty vector and HT-29,respectively,into the mid-posterior part of the left armpit of the nude mice. The rate of tumor growth inhibition was calculated by measuring the volume and weight of the CCTTs in the nude mice. The mRNA and protein expressions of survivin,Cytc,Smac and Caspase-9 in the tumor tissue were detected by real time PCR and immunohistochemistry,respectively,and the apoptosis rate of the CCTT cells was detected by TUNEL. Results The model of CCTT was successfully established in the nude mice. Com-pared with the empty vector and blank transfection groups,the mice in the double-gene,survivin shRNA and APC groups showed sig-nificantly decreased average volume and weight of the tumor tissue (P<0.05) but increased inhibition rate of its volume and weight (P<0.05). In comparison with the survivin shRNA and APC groups,the double-gene group exhibited remarkably decreased average volume and weight of the tumor tissue (P<0.05) but increased inhibition rate of its volume and weight (P<0.05). The mRNA and pro-tein expressions of survivin were significantly lower while those of Cytc,Smac and Caspase-9 markedly higher in the double-gene,sur-vivin shRNA and APC groups than in the empty vector and blank transfection groups (P<0.05),the former even lower (P<0.05) and the latter even higher in the double-gene than in the survivin shRNA and APC groups (P<0.05). The apoptosis rate of the CCTT cells was significantly increased in the double-gene ([56.78±3.04]%),survivin shRNA ([33.61±2.02]%) and APC groups ([30.16± 1.72]%) as compared with the empty vector ([10.05±0.42]%) and blank transfection groups ([9.87±0.30])% (P<0.05),even higher in the double-gene group than in the survivin shRNA and APC groups (P<0.05). Conclusion The Survivin shRNA-APC double-gene may induce apoptosis of colon cancer transplanted tumor cells by down-regulating the expression of the apoptosis inhibitor survivin,upregulating the expressions of Cytc,Smac and Caspase-9,and suppressing the growth of the colon transplanted tumor,with more significant abilities than a single gene in regulating apoptosis-related factors,inducing cell apoptosis and inhibiting the growth of the transplanted tumor.

Dynamic visual analysis of neutrophils chemotaxis in peritoneal cavity of schizophrenic model in mice / 北京大学学报(医学版)

OBJECTIVE@#To compare the differences of neutrophils chemotaxis ability in peritoneal cavity between normal rats and schizopherenic rats with cell dynamic visualization system.@*METHODS@#In the study,18 healthy Kunming rats were randomly divided into 3 groups which were control group (n=6), 0.3 mg/kg MK-801 treatment group (n=6), 0.6 mg/kg dizocilpine maleate (MK-801) treatment group(n=6), extracted neutrophils separately, and observed the morphology and counted under a microscope. Each group of cells was divided into two parts for chemotactic experiment, called chemokine agent treatment group and no chemokine agent treatment group respectively, indicating control 1, 0.3 mg/kg MK-801 treatment 1,0.6 mg/kg MK-801 treatment 1 and control 2, 0.3 mg/kg MK-801 treatment 2,0.6 mg/kg MK-801 treatment 2. The dynamic migration of cells was recorded using the NIS-Elements software, and TAXIScan Analyzer 2 software was used to select 30 cells (n=30) in each group of cells and analyze cells migration trajectory, speed and distance, and use pair test and One-Way analysis of variance for statistical analysis.@*RESULTS@#The number of neutrophils in control group, 0.3 mg/kg MK-801 treatment group and 0.6 mg/kg MK-801 treatment group were(1.00±0.03)×104/mL,(0.05±0.02)×104/mL,(0.32±0.01)×104/mL respectively, the differences of results were statistically significant(P<0.05).Under the effect of chemotactic agent,the directional migration capability of neutrophils in control group 1, 0.3 mg/kg MK-801 treatment group 1 and 0.6 mg/kg MK-801 treatment group 1 were(0.85±0.11) radian,(1.00±0.11) radian,(0.96±0.10) radian respectively (P<0.05); the migration velocities of neutrophils were (0.09±0.02) μm/s,(0.12±0.01) μm/s,(0.14±0.01) μm/s respectively (P<0.05);the migration distances of neutrophils were (94.26±0.02) μm,(134.61±0.01) μm,(156.19±0.01) μm respectively(P<0.05).@*CONCLUSION@#Compared with neutrophils in peritoneal cavity of control group, the neutrophils in peritoneal cavity of schizophrenic rats have stronger chemotactic movement ability.

Application of anoptomagnetic probe Gd-DO3A-EA-FITC in imaging and analyzing the brain interstitial space / 北京大学学报(医学版)

OBJECTIVE@#To investigate the application of the optical magnetic bimodal molecular probe Gd-DO3A-ethylthiouret-fluorescein isothiocyanate (Gd -DO3A-EA-FITC) in brain tissue imaging and brain interstitial space (ISS).@*METHODS@#In the study, 24 male SD rats were randomly divided into 3 groups, including magnetic probe group (n=6), optical probe group (n=6) and optical magnetic bimodal probe group (n=12), then the optical magnetic bimodal probe group was divided equally into magnetic probe subgroup (n=6) and optical probe subgroup (n=6). Referencing the brain stereotaxic atlas, the coronal globus pallidus as center level, the probes including gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA), fluorescein isothiocyanate (FITC) and Gd-DO3A-EA-FITC of 2 μL (10 mmol/L) were injected into the caudate nucleus respectively, magnetic resonance imaging (MRI) was performed in the magnetic probe group and magnetic probe subgroup to image the dynamic diffusion and distribution of the probes in the brain ISS, a self-developed brain ISS image processing system was used to measure the diffusion coefficient, clearance, volume fraction and half-time in these two groups. Laser scanning confocal microscope (LSCM) was performed in vitro in the optical probe group and optical probe subgroup for fluorescence imaging at the time points 2 hours after the injection of the probe, and the distribution in the oblique sagittal slice was compared with the result of the first two groups.@*RESULTS@#For the magnetic probe group and magnetic probe subgroup, there were the same imaging results between the probes of Gd-DTPA and Gd-DO3A-EA-FITC. The diffusion parameters of Gd-DTPA and Gd-DO3A-EA-FITC were as follows: the average diffusion coefficients [(3.31±0.11)×10-4 mm2/s vs. (3.37±0.15)×10-4 mm2/s, t=0.942, P=0.360], the clearance [(3.04±0.37) mmol/L vs. (2.90±0.51) mmol/L, t=0.640, P=0.531], the volume fractions (17.18%±0.14% vs. 17.31%±0.15%, t=1.961, P=0.068), the half-time [(86.58±3.31) min vs. (84.61±2.38) min, t=1.412, P=0.177], the diffusion areas [(23.25±0.68) mm2 vs. (22.71±1.00) mm2, t=1.100, P=0.297]. The statistical analysis of each brain was made by t test, and the diffusion parameters were not statistically significant. Moreover, for the optical probe group and optical probe subgroup, the diffusion area of Gd-DO3A-EA-FITC [(22.61±1.16) mm2] was slightly larger than that of FITC [(22.10±1.29) mm2], the statistical analysis of each brain was made by t test, and the diffusion parameters were not statistically significant (t=0.713, P=0.492).@*CONCLUSION@#Gd-DO3A-EA-FITC shows the same imaging results as the traditional GD-DTPA, and it can be used in measuring brain ISS.

Transcutaneous electrical acupoint stimulation on inflammatory response and intestinal permeability in perioperative period of laparoscopic intestinal surgery / 中国针灸

OBJECTIVE@#To observe the effect of transcutaneous electrical acupoint stimulation (TEAS) on the inflammatory response and intestinal permeability after laparoscopic radical surgery for colon cancer, so as to explore the protective mechanism for perioperative organs.@*METHODS@#Sixty patients with laparoscopic intestinal resection were randomly assigned into an observation group and a control group, 30 cases in each group. TEAS at Zusanli (ST 36), Neiguan (PC 6) and Hegu (LI 4) was used in the observation group on the day of operation and within 3 days after surgery, 2 Hz, within 2 mA. The interventions in the control group were the same as those in the observation group, except opening the current source. Central venous blood was collected before TEAS and 12 h, 24 h, 48 h, 72 h after surgery. The level of the inflammatory indexes of tumor necrosis factor-α(TNF-α), interleukin-1 (IL-1), interleukin-6 (IL-6) and intestinal mucosal barrier response indexes of endotoxin (LPS) and D-lactate were detected.@*RESULTS@#Compared with the control group, the levels of IL-1, IL-6 and TNF-α decreased in the observation group 24 h, 48 h and 72 h after surgery (all 0.05).@*CONCLUSION@#TEAS can reduce the inflammatory reaction after laparoscopic radical surgery for colon cancer, but has no effect on the intestinal permeability.

Effect of transcutaneous electrical acupoint stimulation on gastrointestinal function in perioperative period of laparoscopic intestinal surgery / 临床麻醉学杂志

Objective To observe the effect of transcutaneous electrical acupoint stimulation (TEAS) on the recovery of gastrointestinal function after laparoscopic intestinal operation.Methods Sixty patients of the selective laparoscopic intestinal resection, 28 males and 32 females, aged 18-65 years, ASA physical status Ⅰ or Ⅱ, were enrolled and randomly allocated into two groups: TEAS group and control group, 30 in each group.Patients in TEAS group accepted transcutaneous electrical acupoint stimulation treatment at Neiguan, Hegu, Zusanli points from the time before induction of anesthesia to 3 days after surgery, and patients in the control group were treated with transcutaneous electrical acupoint stimulation, but the electrode pads were just attached on the related points with no electric stimulation.Plasma motilin concentrations preoperatively, postoperatively 12, 24, 48 and 72 h were measured in the two groups.The recovery time of intestinal peristalsis, anal flatus time, in-hospital time and the incidence of nausea and vomiting within 3 days after operation were observed.Results Compared with the control group, serum motilin concentration postoperative 24 h increased significantly [(218.5±52.3) pg/ml vs (141.8±45.8) pg/ml, P<0.05], the time of intestinal peristalsis recovery [(19.4±3.2) h vs (29.6±7.8) h, P<0.05] and flatus [(23.2±4.7) h vs (36.5±8.9) h, P<0.05] were shorter, the incidence of postoperative nausea and vomiting within 3 days after operation decreased significantly in TEAS group (16.7% vs 36.7%, P<0.05).There was no statistically significant difference of the in-hospital time between the two groups.Conclusion TEAS can promote the recovery of gastrointestinal function in patients undergoing laparoscopic intestinal surgery.

Change of PD-L1 in Peripheral Blood Mononuclear Cells of Patients with Primary Biliary Cirrhosis and Its Significance / 现代检验医学杂志

Objective To explore the expression of PD-L1 in peripheral blood mononuclear cells (PBMC) from patients with primary biliary cirrhosis (PBC) and its significance.Methods Peripheral blood of 23 healthy individuals and 23 patients with PBC diagnosed in Changhai Hospital was collected during February 2013 to December 2015.PBMC was separated by density gradient centrifugation,then cultured.The mRNA expression of PD-L1 in PBMC was examined by real-time fluorescent quantitative PCR (qRT-PCR),the expression of cytokines such as TNF-α,TGF-β,IL-23,IL-17 in medium was examined by enzyme-linked immunosorbent assay (ELISA).Two independent samples' t test was used to compare variables in two groups.Pearson correlative coefficient was used to reveal the relationship between two variables.Results mRNA expression of PD-L1 in experiment and control groups was 0.23±0.08 vs 1.27±0.40 (t=12.23,P＜0.000 1) and there was statistical significance.Pearson correlation analysis revealed that PD-L1 was negatively correlated with IL-23(r=-0.531,P =0.009) with difference statistically.Conclusion PD-L1 might involve in the development of PBC,and could be the potential biomarker for predicting and treating this disease.

Clinical Study on the Effect of Pre-intervention with Electroacupuncture on Oxidative Stress and Inflammatory Reaction in Hepatic Ischemia-reperfusion / 上海针灸杂志

Objective To observe the effect of pre-intervention with electroacupuncture on oxidative stress (OS) and inflammatory reaction after hepatectomy under portal occlusion.Method Thirty patients going to receive hepatectomy under Pringle's portal occlusion were randomized into an electroacupuncture group and a sham electroacupuncture group, 15 cases each. Three days prior to the surgery, the patients started to receive corresponding acupuncture or sham electroacupuncture treatment by selecting bilateral Riyue (GB24), Qichong (ST30) and Yanglingquan (GB34), once a day. The anesthesia method was general (inhalational and intravenous) anesthesia. Central venous blood was drawn before the surgery, and respectively 6 h, 24 h, 48 h and 72 h after the surgery to examine the OS-relatedparameters, superoxide dismutase (SOD) and malondialdehyde (MDA), as well as the inflammatory mediators, including tumor necrosis factor-α(TNF-α), interleukin (IL)-1, IL-6, and IL-10.Result Compared to the sham electroacupuncture group, the SOD activity significantly increased (241±11.4 U/mL,180±12.6 U/mL,P0.05).Conclusion Pre-intervention with electroacupuncture can up-regulate the activity of SOD and down-regulate the level of MDA in patients after hepatic portal occlusion, and thus partially reduce OS, but it doesn't shownoticeable effecton inflammatory response.

Elevated Expression of CD93 in Serous Exosomes from Patients with Cryptococcal Meningitis and Its Significance / 现代检验医学杂志

Objective To examine the expression of CD93 in serous exosomes from patients with cryptococcal meningitis,fur-ther explore its clinical significance.Methods The 38 experimental serum samples were from patients who received the diag-nosis with cryptococcal meningitis in Changhai Hospital and Changzheng Hospital in Shanghai from November 2012 to De-cember 2016.The diagnosis standardization was that the cerebrospinal fluid dyeing was positive or the culturing was posi-tive.The 38 controls were collected from the health individuals examined at the same time.The exosomes in serum was sepa-rated by the ultracentrifuge method.Magnetic bead-capture combined with the flow cytometry method was used to identify antigens on the surface of exosomes.The protein level of cytokines in serum was quantified by ELISA method.The compari-son of tested values from experimental and controlled groups was measured by two independent samples't test,and the cor-relation between two variates was showed by Pearson coefficient.Results The results of flow cytometry showed that the ex-pression of CD93 on exosomes in the serum samples from experimental group was higher than that in the controlled group, which was(79.11±19.31 vs 23.98±6.56)%,with difference in statistics(t=16.66,P<0.000 1).The expression levels of IFN-γin serum samplefrom experimental and control groups were(39.78±10.77 vs 58.98±16.99)pg/ml,with differ-ence in statistics(t=5.884,P<0.000 1).The expression levels of IL-17 in serum sample from experimental and control groups were(16.32±4.03 vs 3.11±0.87)pg/ml,with difference in statistics(t=19.75,P<0.000 1).The expression lev-els of IL-1β in serum sample from experimental and control groups were(57.12 ± 12.98 vs 13.45 ± 4.78)pg/ml,with difference in statistics(t=19.46,P<0.000 1).The percentage of CD93+exosomes was positively correlative with IFN-γ, IL-17,IL-1β(r=0.488,0.456,0.532,P<0.01)in serum samples from the experimental group and there was statistical difference.Conclusion In the disease process of cryptococcal meningitis,the expression of CD93 on exosomes might involve in the diseases'processes by affecting the CD4+T cells.

Protection of Electro-acupuncture for Gastric Mucosa of Patients Undergoing Endoscopic Sinus Surgery / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To evaluate the effect of electro-acupuncture (EA) on gastric mucosal oxygenation and systemic inflammatory response in patients undergoing endoscopic sinus surgery with controlled hypotension (CH), and to explore its protective effect on gastric mucosa.</p><p><b>METHODS</b>Fifty-four patients, 18-65 years old, grade I-II of American Society of Anesthesiology (ASA), who were scheduled for endoscopic sinus surgery were randomly assigned to two groups, group A (general anesthesia group) and group B (general anesthesia combined EA anesthesia group), 27 in each group. Conrolled hypotension was executed during operation, and mean arterial pressure (MAP) was maintained at 55-65 mmHg. After tracheal intubation gastric tesiometer catheter was indwelled through nasal cavity or oral cavity. After successful indwelling, it was connected with gastric mucosa monitoring mode of multifunctional parameters monitor. Patients' MAP and heart rate (HR), pHi, partial pressure of carbon dioxide (PgCO2), arterial partial pressure of carbon dioxide (Pg-aCO2) and endtidal pressure of carbon dioxide (Pg-etCO2) were measured and recorded at T, (immediately before induced hypotension), T, (20 min following induced hypotension to target MAP), T2 (40 min following induced hypotension to target MAP), T3 (20 min after ending induced hypotension), and T4(40 min after ending induced hypotension). Blood samples were intravenously collected, TNF-alpha, IL-1, and IL-6 were detected by ELISA 24 h before operation, during operation (T3), and 24 h after operation.</p><p><b>RESULTS</b>After hypotension was induced, Pg-CO2, Pg-aCO2 and Pg-etCO2 increased significantly (P < 0.01, P < 0.05), while pHi decreased significantly (P < 0.01) in both groups at T1-T4 than those at T0. During T1-T4, PgCO2, Pg-aCO2, and Pg-etCO2 were higher (P < 0.01, P < 0.05), while pHi was lower in group A than in group B (P < 0.01). Furthermore, TNF-alpha, IL-1, and IL-6 increased significantly in both groups during operation and 24 h after operation, when compared with those 24 h before operation (P < 0.01, P < 0.05). TNF-alpha and IL-1 in group A were higher than those in group B (P < 0.05) during operation and 24 h after operation, but with no significant difference in the plasma concentration of IL-6 (P > 0.05).</p><p><b>CONCLUSION</b>EA exerted obvious protective effect of gastric mucosal injury in endoscopic sinus surgery with controlled hypotension, which might be achieved by increasing gastric mucosal blood flow, maintaining oxygen supply and demand, inhibiting inflammatory response, and alleviating injury of gastric mucosal barrier.</p>

Clinical examples of professor LI Zhi-dao's "tonifying three qi" acupuncture method / 中国针灸

Professor LI Zhi-dao, according to acupoint selection of syndrome differentiation in TCM basic theory, concluded a new therapy, namely "tonifying three qi" that is mainly based on three acupoints in the Conception Vessel. This method is consisted of Danzhong (CV 17), Zhongwan (CV 12) and Qihai (CV 6) in the Conception Vessel, which could successively nourish clear qi, stomach qi and original qi. In clinic, according to the severity of symptoms of three qi, the acupoints are selected flexibly, which could respectively treat deficiency of heart-lung qi, deficiency of stomach-spleen qi and deficiency of original qi. Some examples are also given in the article.

Effect of application of acupuncture-anesthetic composite anesthesia on hysteroscopic surgery: a clinical study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the analgesic effect and safety of acupuncture-anesthetic composite anesthesia (AACA) in hysteroscopic surgery.</p><p><b>METHODS</b>Totally 93 patients undergoing hysteroscopic surgery were randomly assigned to the intravenous anesthesia group (A group, 30 cases), the AACA group (B group, 32 cases), and the acupuncture combined with intravenous anesthesia group (C group, 31 cases). Patients in Group A were anesthetized by sufentanil combined propofol. Those in Group B were anesthetized by sufentanil combined acupuncture. Those in Group C were anesthetized by sufentanil, propofol combined acupuncture. Yinlian and Ququan (LR8) were needled for patients in Group B and C. The peri-operative mean arterial pressure (MAP), heart rate (HR), and oxygen saturation (SpO2), the surgical time, the recovery time, the sufentanil and propofol dosages, adverse anesthesia reactions were observed. Meanwhile, the OAA/S score, Ramsay sedation score, and Visual Analogue Score (VAS) were also measured.</p><p><b>RESULTS</b>Compared with Group A and C, patients in Group B were awake, with obvious increased OAA/S score (P < 0.01). Ramsay sedation score was significantly lower (P < 0.01).The MAP and HR were elevated (P < 0.05). The patient case of SpO2 less than 85% during the operation decreased (P < 0.05). The incidence of postoperative dizziness was reduced (P < 0.05). Compared with Group A, the propofol consumption decreased in Group C (P < 0.05). There was no statistical difference in the operation time, the sufentanil dosage, VAS score, the incidence of postoperative nause- a and vomiting among the three groups (P > 0.05).</p><p><b>CONCLUSIONS</b>The patients were awake in AACA. The intraoperative sedation was better than that obtained by intravenous anesthesia. But the analgesic effect was similar to that obtained by intravenous anesthesia.</p>

Hydroxycamptothecin-induced apoptosis of human Tenon capsule fibroblast and its mechanism / 中华实验眼科杂志

Background The hyperplasia of human Tenon capsule fibroblasts (HTFs) is a common cause of filtering surgery failure in glaucomous eye.Researches demonstrated that hydroxycamptothecin is a cell cycle arresting drug and induce apoptosis of cancer and fibroblasts.However,its mechanism is currently less understood.Objective This study was to investigate whether hydroxycamptothecin induce the apoptosis of HTFs and explore the possible mechanism.Methods Human Tenon capsule tissue was obtained from EyeBank of Jiangsu Province Hospital.HTFs were cultured using explant method in vivo and passaged in DMEM containing 10％ FBS.The cells were identified using vimentin and keratin by immunochemistry,and the cells of generation 3-6 in the logarithmic growth phase were used in the experiment.The cells were incubated with 0.01,0.05 or 0.10 g/L hydroxycamptothecin for 5 minutes respectively,and the cells without any hydroxycamptothecin were served as the control group.Cell viability then was assessed by cell counting kit-8 (CCK8) for the optimal inhibition concentration.The cells were treated by 0.10 g/L hydroxycamptothecin for 24 hours,and the apoptotic rate of the cells were assayed with annexin V/PI double-staining.Mitochondrial membrane potential of HTFs was assessed using JC-1 staining.The expressions of caspase-3,caspase-9 and cytochrome C (cyt C) in mitochondria and cytoplasm of HTFs were detected by Western blot.Results The proliferative value (A450) of the HTFs 0,0.01,0.05,0.10 g/L was 0.9716±0.0608,0.8035 ± 0.0346,0.7048 ±0.0446,0.6265 ±0.0286,with a significant difference (F =26.372,P =0.002).A450 of HTFs in the 0.01,0.05,0.10 g/L groups was significantly lower than the control group (P＜0.05),with the lowest A450 value in the 0.10 g/L group.The apoptotic percentage of HTFs was (18.72±1.41)％,in the 0.10 g/L hydroxycamptothecin group and that of the control group was (3.67 ±0.36)％,showing a significant difference between them (t =-10.374,P=0.001).The expression intensity of caspase-3 and caspase-9 protein in HTFs was higher in the 0.10 g/L hydroxycamptothecin group than that in the control group.JC-1 staining showed that the green fluorescence of the monomer JC-1 in cytoplasm was stronger in the 0.10 g/L hydroxycamptothecin group than that in the control group,but the red fluorescence of the polymer JC-1 in the 0.10 g/L hydroxycamptothecin group was weaker than that in the control group.The grey scale of cyt C protein in HTFs in mitochondrion was 0.0605±0.0022 in the 0.10 g/L hydroxycamptothecin group,showing a significant increase in comparison with 0.0301 ±0.0016 of the control group (t=4.865,P=0.014).However,the grey scale of cyt C protein in cytoplasm was declined in the 0.10 g/L hydroxycamptothecin group than that in the control group (0.0605 ±0.0022 vs.0.0301 ±0.0016) (t =-11.177,P =0.001).Conclusions Hydroxycamptothecin can induce the apoptosis of HTFs through activating the mitochondrial apoptosis pathway.

Impacts of electroacupuncture at different frequencies on the postoperative nausea and vomiting of patients with laparoscopic surgery / 中国针灸

<p><b>OBJECTIVE</b>To assess the impacts on postoperative nausea and vomiting of the patients with laparoscopic surgery undergoing the general anesthesia by electroacupuncture (EA) at different frequencies at bilateral Neiguan (PC 6).</p><p><b>METHODS</b>One hundred and twenty female patients with laparoscopic surgery undergoing the general anesthesia were randomized into 4 groups: 2 Hz EA treatment group (group A), 2 Hz/100 Hz EA treatment group (group B), 100 Hz EA treatment group (group C) and a control group (group D), 30 cases in each one. The same anesthetic program was adopted in each group. For the patients in A, B and C groups, in the first half a hour undergoing the conventional drug anesthesia, acupuncture was applied to bilateral Neiguan (PC 6). After arrival of qi, HANS-200A electroacupuncture apparatus was connected, at the frequency as 2 Hz, 2 Hz/100 Hz and 100 Hz separately. The electric stimulation was discontinued at the end of surgery. The life physical signs after surgery, the occurrence of postoperative nausea and vomiting and the severity grades of postoperative nausea and vomiting were observed and recorded for the patients in each group.</p><p><b>RESULTS</b>The differences in the postoperative 1 h heart rate, mean arterial pressure (MAP), finger pulse oxygen saturation and respiratory frequency were not significant statistically in comparison of the patients among groups (all P > 0.05). The incidence of postoperative nausea and vomiting of the patients in group B was lower apparently than that in A, C and D groups [10.0% (3/30) vs 30.0% (9/30), 36.7% (11/30), 53.3% (16/30), all P < 0.01]. The severity of postoperative nausea and vomiting of the patients in group B was lower apparently than that in the rest three groups (all P < 0.01).</p><p><b>CONCLUSION</b>The electroacupuncture at 2 Hz/100 Hz reduces effectively the incidence and the severity of postopera tive nausea and vomiting of the patients with laparoscopic surgery undergoing the general anesthesia.</p>