Objective To investigate the effects of adenosine monophosphate-activated protein kinase (AMPK) on high-mobility group box 1 (HMGB1) release from PC12 cells after oxygen-glucose deprivation and reoxygenation (OGD/R) and its mediated inflammatory response in BV2 cells.Methods PC12 and BV2 cells were cultured,respectively.The PC12 cells were used to induce a model of oxygen glucose deprivation for 12 h and reoxygenation for 24 h.After giving 5-aminoimidazole-4-carboxamide (AICAR) 5,50 and 100 μmol/L as well as Compound C 0.1,1 and 10 μmol/L activation or inhibition of AMPK phosphorylation,respectively,methyl thiazolyl tetrazolium (MTT) was used to detect the PC12 cell activity.Enzyme-linked immunosorbent assay was used to detect the HMGB1 release level in the PC12 cell culture media.After OGD/R in each group,the PC12 culture media were acted on normal cultured BV2 cells for 24 h respectively.Westem blotting and Enzyme-linked immunosorbent assay were used to detect the NFκB inhibitory protein (inhibitor of NFκB,IκB) phosphorylation level and TNF-α release level in BV2 cells,respectively.Results After OGD/R,the PC12 cell activity was decreased significantly (68.84％±6.60％ vs.100.04％ ± 8.82％;P ＜ 0.01);the AMPK phosphorylation level was increased significantly (1.95 ±0.39 vs.1.00 ±0.20;P＜0.05),and the extracellular HMGB1 release was increased significantly (287.66 ± 26.42 pg/μl vs.53.05 ± 9.11 pg/μl;P ＜ 0.01).Compared with the OGD/R group,AICAR 100 μmol/L significantly increased the survival rate of PC12 cell after OGD/R (78.6％ ± 3.75％ vs.68.84％ ± 6.60％;P ＜ 0.05),promoted AMPK phosphorylation (3.32 ± 0.66 vs.1.95 ± 0.39;P ＜ 0.01),and reduce the release of extracellular HMGB1 (164.06 ± 12.77 pg/μl vs.287.66 ± 26.42 pg/μl;P ＜0.01).In contrast,Compound C 10 μmol/L significantly reduced the cell survival rate of PC12 (40.44％ ±3.79％ vs.68.84％ ±6.60％;P ＜0.01),inhibited AMPK phosphorylation (1.07 ± 0.21 vs.1.95 ± 0.39;P＜0.05),and increased the release of HMGB1 (337.97 ± 18.9 pg/μlvs.287.66 ± 26.42 pg/μl;P＜0.01).The conditioned medium from the AICAR 100 μmol/L group significantly inhibited IκB phosphorylation (1.68 ±0.51 vs.3.09 ± 0.10;P ＜ 0.05) and reduced the release of TNF-α (669.53 ±38.58 pg/μlvs.841.76 ± 45.82 pg/μl;P＜ 0.05) in BV2 cells.The conditioned medium from the compound C 10 μmol/L group significantly promoted IκB phosphorylation (4.98 ± 1.24 vs.3.09 ± 0.10;P ＜0.01) and increased the release of TNF-α (1 035.32 ± 128.06 pg/μl vs.841.76 ± 45.82 pg/μl;P ＜0.05) in BV2 cells.Conclusions Promoting AMPK phosphorylation activation may reduce the release of HMGB1 from PC12 cells after OGD/R,and inhibit its mediated NF-κB inflammatory pathway and reduce the release of TNF-αin BV2 cells,and thus reducing neuroinflammatory injury.On the contrary,inhibiting AMPK phosphorylation may promote the release of HMGB1 from PC12 cells after OGD/R and aggravate its mediated inflammatory reaction in BV2 cells.

Objective Discuss therapy problems in NHL patients with diabetics. Methods 40 NHL patients with diabetics are treated with chemotherapy containing prednisone. Compare the blood sugar before using prednisone with the one after that, regulate medicines of reducing glucose and at the same time observe the syndromes. Results Limosis blood sugar of patients just in hospital is (6.1?1.3) mmol/L, and after chemotherapy it becomes (6.5?1.2) mmol/L. There is no significant difference between them. Conclusions On the basis of controlling blood sugar effectively, patients with NHL combining diabetics can use prednisone as normal.

Objective: To investigate the clinicopathological characteristics of SMARCA4- deficient thoracic sarcomas. Methods: The clinical features and CT scans of SMARCA4-deficient thoracic sarcomas (n=5) diagnosed at Fudan University Cancer Hospital from December 2016 to October 2018 were reviewed. Hematoxylin-eosin staining, immunohistochemistry and targeted next generation sequencing were performed in available cases along with a literature review. Results: All 5 patients were males with age ranging from 32 to 65 years (average 54 years; median 61 years). Four patients were smokers except one with unknown smoking history. The average maximum diameter of tumor was 5.6 cm. Tumor primary sites included thoracic wall,thoracic cavity,lung and mediastinum. Histologically,tumor cells formed solid sheets or anastomosing islands with brisk mitotic figures accompanying with large areas of necrosis. Three cases focally exhibited rhabdoid morphology and vesicular chromatin. Immunohistochemically, SMARCA4, SMARCA2 and Claudin-4 were negative in all cases and all tumors demonstrated SOX2 and SMARCB1 nuclear positive staining. Among 3 cases analyzed by targeted next generation sequencing, concurrent SMARCA4 and p53 mutation was detected in all three cases. Mutations of STK11, ERBB4, NF2, GNAS, MTOR,MET and FGFR1 amplification were also detected among the three cases. The follow-up information was available in all 5 cases. Two patients died of the tumor. One relapsed multiple times after surgeries but was alive with disease. Two patients received radical excisions without relapse. Conclusions SMARCA4-deficient thoracic sarcoma is a rare but highly-aggressive tumor with dismal prognosis. The tumor is featured by rhabdoid morphology histologically and distinctive immunohistochemical and molecular phenotype.

Objective To analyze the electromyogram (EMG) changes of patients with acute flaccid paralysis (AFP) in Hetian Xinjiang.Methods Twenty-six patients with acute poliogmyelitis,admitted to our hospital from July 2011 to September 2012,were chosen in our study; their clinical data were analyzed,and they all underwent EMG examination.Results Most patients had contact with respiratory virus or digestive virus prior to illness onset.Most of patients had flaccid paralysis,muscle strength between 0 and 2 grades,tendon reflex being abated and no sensory disturbance.Fourteen patients had positive blood stool.Motor nerve conduction velocity (MCV) was abnormal,H-reflex was decreased or not educed and sensory nerve conduction velocity was normal in lower limbs of the 26 patients; EMG showed myogenic abnormality.Nerve conduction velocity was normal or low and Fwave was normal or decreased in upper limbs; EMG showed neurogenic abnormality in five of the 26 patients.Poor prognosis was concluded in all the patients.Conclusion EMG has important diagnosis value in AFP induced by polio wild-type I.

Flavonoids have a variety of biological activities and have important applications in food, medicine, cosmetics, and many other fields. Naringenin is a platform chemical for the biosynthesis of many important flavonoids. Ubiquitination plays a pivotal role in the post-translational modification of proteins and participates in the regulation of cellular activities. Ubiquitinated proteins can be degraded by the ubiquitin-protease system, which is important for maintaining the physiological activities of cells, and may also exert a significant impact on the expression of exogenous proteins. In this study, a real-time in-situ detection system for ubiquitination modification has been established in Saccharomyces cerevisiae by using a fluorescence bimolecular complementation approach. The ubiquitination level of protein was characterized by fluorescence intensity. By using the approach, the potential ubiquitination sites of proteins involved in the naringenin biosynthesis pathway have been obtained. The lysine residues of the relevant ubiquitination sites were mutated to arginine to reduce the ubiquitination level. The mutants of tyrosine ammonia-lyase (FjTAL) and chalcone synthase (SjCHS, SmCHS) showed decreased fluorescence, suggested that a decreased ubiquitination level. After fermentation verification, the S. cerevisiae expressing tyrosine ammonia-lyase FjTAL mutant FjTAL-K487R accumulated 74.2 mg/L p-coumaric acid at 72 h, which was 32.3% higher than that of the original FjTAL. The strains expressing chalcone synthase mutants showed no significant change in the titer of naringenin. The results showed that mutation of the potential ubiquitination sites of proteins involved in the naringenin biosynthesis pathway could increase the titer of p-coumaric acid and have positive effect on naringenin biosynthesis.
Biosynthetic Pathways ; Flavanones/metabolism* ; Saccharomyces cerevisiae/metabolism* ; Ubiquitination

Objective:To investigate the differences between the expression levels of ribosomal protein L32 (RPL32) in human breast cancer tissue and normal breast tissue and the effects on the proliferation of breast cancer cells.Methods:Paraffin samples of breast cancer tissues and adjacent tissues (more than 3 cm from the tumor margin) were collected from 56 breast cancer patients in the Department of Thyroid and Breast Surgery of the First People's Hospital of Shangqiu City from July 2020 to May 2022. The expression of RPL32 in 56 breast cancer patients and their corresponding paracancer tissues was detected by immunohistochemistry. MCF7 cells were divided into experimental group (ribosomal protein L32, RPL32) and control group (negative control, NC). MCF7 cells in experimental group were transfected with RPL32-siRNA vector, while MCF7 cells in control group were transfected with scramble siRNA vector. RPL32 mRNA content in each group was detected by RT-PCR. The expressions of RPL32 and P53 in the experimental group and control group were detected by western blot. The proliferative ability of cells in each group was detected by CCK8 assay. The clonogenesis ability of each group of cells was detected by clone formation experiment.Results:The positive rate of RPL32 in breast cancer patients was 8.93% (5/56), and the expression rate of RPL32 in paracancer tissues was 78.57% (44/56). The expression rate of RPL32 in breast cancer patients was significantly higher than that in paracancer tissues, with statistical significance ( P=0.007). After transfection with siRNA vector, the mRNA content of RPL32 in MCF7 cells of experimental group and control group decreased, and the protein expression level of RPL32 was 1.09±0.21 and 0.40±0.11, respectively. The expression levels of P53 protein were 1.24±0.32 and 0.37±0.09, respectively. The absorbance of CCK8 at 120 h was 1.11±0.24 and 2.19±0.28, respectively, and the proliferation ability of MCF7 cells in the experimental group was significantly decreased ( P=0.043). The results of clone formation experiment showed that the cell clone formation rate of the experimental group and the control group was (21.11±3.46) % and (58.75±4.29) %, respectively, and the cell clone formation of the experimental group was decreased ( P=0.026) . Conclusions:The expression of RPL32 is significantly increased in breast cancer, which may be related to the malignant degree of breast cancer. Inhibition of RPL32 expression in breast cancer cells affects its proliferation ability.

Objective:To analyze the protective effect of neurotracheal catheter monitoring on recurrent laryngeal nerve in patients with bilateral cervical lymph node dissection of thyroid cancer.Methods:The clinical data of 92 patients undergoing bilateral cervical lymph node dissection for thyroid cancer at the First People's Hospital of Shangqiu from Jul 2019 to Aug 2022 were retrospectively analyzed. Patients were divided into control group (routine exposure, 52 cases) and study group (intraoperative neurotrachatic catheter monitoring, 40 cases) The general data, perioperative indicators, parathyroid function, vocal cord function, voice disorder index, quality of life and postoperative complications were compared between the two groups.Results:The intraoperative blood loss in the study group [(12.3±3.3) ml] was less than that in the control group [(16.9±4.0) ml]. The exposure time [(8.7±2.6) min], operation time [(4.0±0.8) h] and postoperative hospitalization time [(3.2±0.9) d] were shorter than those of control group [(14.4±3.6) min, (4.5±1.2) h, (5.7±1.3) d] ( t=5.770, 8.391, 2.387, 10.853, all P<0.05);Amplitude perturbation (4.85%± 0.58%), fundamental frequency perturbation (0.28%±0.17%), standardized noise energy [(-20.3±4.4) dB], VHI-10 score [(1.6±0.5) score], Quality of life scale for cancer patients 1 month after surgery the TG-V4 score [(43.7±4.8) points] was lower than that of control group [(3.05%±0.54%), (0.42%±0.16%), (-14.6±3.3) dB, (3.3±0.4) points, (50.4±5.6) points]. The harmonic noise ratio [(24.9±4.1) dB] was higher than that of the control group [(20.3±4.4) dB] ( t=10.446, 4.049, 7.036, 19.076, 6.116, 5.144, all P<0.001);The incidence of postoperative complications in the study group (8%) was lower than that in the control group (23%) ( χ2=4.020, P=0.045). Conclusion:Neurotracheal catheter monitoring during bilateral cervical lymph node dissection for thyroid cancer can improve vocal cord function, reduce recurrent laryngeal nerve injury, reduce voice disturbance and improve quality of life.

Blood brain barrier (BBB) injury is the main pathological manifestation of many neurological diseases. Glycocalyx is the gel layer covering the lumen side of vascular endothelial cells, which plays an important role in regulating BBB function. However, glycocalyx is very fragile and easily damaged in various neurological diseases, leading to BBB destruction. This article focuses on the potential role of glycocalyx in cerebrovascular disease, the possible mechanisms related to glycocalyx and BBB injury, and explores the potential therapeutic strategies for protecting and restoring endothelial glycocalyx.

Objective To investigate the clinicopathological characteristics of SMARCA4?deficient thoracic sarcomas. Methods The clinical features and CT scans of SMARCA4?deficient thoracic sarcomas (n=5) diagnosed at Fudan University Cancer Hospital from December 2016 to October 2018 were reviewed. Hematoxylin?eosin staining, immunohistochemistry and targeted next generation sequencing were performed in available cases along with a literature review. Results All 5 patients were males with age ranging from 32 to 65 years (average 54 years; median 61 years). Four patients were smokers except one with unknown smoking history. The average maximum diameter of tumor was 5.6 cm. Tumor primary sites included thoracic wall,thoracic cavity,lung and mediastinum. Histologically,tumor cells formed solid sheets or anastomosing islands with brisk mitotic figures accompanying with large areas of necrosis. Three cases focally exhibited rhabdoid morphology and vesicular chromatin. Immunohistochemically, SMARCA4, SMARCA2 and Claudin?4 were negative in all cases and all tumors demonstrated SOX2 and SMARCB1 nuclear positive staining. Among 3 cases analyzed by targeted next generation sequencing, concurrent SMARCA4 and p53 mutation was detected in all three cases. Mutations of STK11, ERBB4, NF2, GNAS, MTOR,MET and FGFR1 amplification were also detected among the three cases. The follow?up information was available in all 5 cases. Two patients died of the tumor. One relapsed multiple times after surgeries but was alive with disease. Two patients received radical excisions without relapse. Conclusions SMARCA4?deficient thoracic sarcoma is a rare but highly?aggressive tumor with dismal prognosis. The tumor is featured by rhabdoid morphology histologically and distinctive immunohistochemical and molecular phenotype.

ObjectiveTo analyze the effect of O₃ pollution on outpatient visits for respiratory diseases in a district of Shanghai. MethodsWe collected the respiratory disease outpatient data， and atmospheric and meteorological data of from a suburban general hospital in Shanghai from 2015 to 2017. A time-series analysis by generalized additive model was conducted to examine the relationship between O₃ pollution and daily outpatient visits. ResultsThe daily outpatient volume for respiratory diseases was 831. The daily 8 h median concentration of O₃ was 101.04 μg·m^-3. The excess relative risk was 0.461% （95%CI： 0.240%‒0.682%） at lag3. Stratified by gender and age， females， child and the aged had higher risk of respiratory diseases. In the double-pollutant model， PM_2.5 and PM₁₀ increased health effects， while CO reduced health effects. ConclusionThe increase of O₃ concentration can increase the daily outpatient volume of respiratory diseases.