Objective:To study the neuroprotective effect of nobiletin on the symptoms of postoperative cognitive impairment (POCD) induced by sevoflurane inhalation.Methods:Twenty-four aged SD rats (12 female mice and 12 male mice) were divided into three groups randomly: control group ( n=8), surgery group ( n=8) and surgery + nobiletin group ( n=8), with 4 females and 4 males in each group.The rats in surgery group and surgery+ nobiletin group were given normal saline(0.1 ml/10 g, once a day) and nobiletin(100 mg/kg, once a day) intragastrically for 6 weeks.Then the rats were anesthetized by sevoflurane and treated by abdominal exploration surgery, and then continued gavage for 1 week.The rats in control group were given normal saline(0.1 ml/10 g, once a day) intragastrically for 7 weeks without anesthesia or surgery.Sevoflurane inhalation anesthesia and abbreviated laparotomy were not done for control group.Morris water maze and open field experiment were used to measure the memory and cognitive ability and the independent exploration ability respectively.The changes of α-band electroencephalogram (EEG) were recorded by multi-channel physiological signal acquisition and processing system.The concentration of S100β, a marker of neurological impairment was detected by ELISA.Western blot was used to detect the expression level of IBA-1 in microglia.SPSS 20.0 software was used to analyze the data. Results:There were no significant differences in Morris water maze, positioning cruise test and open field test among the groups before operation (all P>0.05). The differences were statistically significant among the groups 7 days after operation (all P<0.05). Compared with the control group (the escape latency, path length and cross platform times were ((20.37±1.11)s, (552.37±14.19)cm, (6.75±0.43)times respectively), the escape latency ((40.87±2.03)s) and path length ((1 258.62±19.53)cm) of rats in surgery group were significantly longer (both P<0.01), and the cross platform times ((2.12±0.33)times) significantly reduced ( P<0.01). The differences between surgery + nobiletin group ((22.37±1.11)s, (584.50±10.90)cm, (6.62±0.48)times) and control group were not significant (all P>0.05). The open field experiment showed that the movement distance, the crossed square number, and activity times in surgery group ((1.78±0.55) m, (4.75±0.50), (14.87±0.33) times) decreased significantly compared with those in control group ((3.73±0.07) m, (11.10±0.78), (51.12±0.78) times, all P<0.01). No significant difference was found between surgery + nobiletin group ((3.76±0.07)m, (10.75±0.66), (50.75±0.43)times) and control group(all P>0.05). Before anesthesia, there was no significant difference in the power ratio of α-band among the three groups ( P>0.05), but the differences during anesthesia and operation were significant ( F=72.58, 101.50, P<0.01). During anesthesia and operation, the power ratio of α-band in anesthesia and in surgery group (2.51±0.04, 2.14±0.03) were significantly lower (both P<0.01) than those in control group (3.49±0.03, 3.49±0.03), while there was no obvious changes (both P>0.05) in the surgery + nobiletin group (3.50±0.04, 3.51±0.04). There were significant differences in Bcl-2 protein expression and caspase 3/7 protein activity among the three groups ( F=5.21, 7.84, P<0.01). Compared with control group (1.00±0.02, 1 557.46±3.63), Bcl-2 of rats in the surgery group(0.40±0.05) were significantly lower and Caspase3/7 expression of surgery group (3 689.58±10.46) was significantly higher (both P<0.01), while the rats in the surgery + nobiletin group had no significant difference in both Bcl-2 level (1.03±0.06) and caspase 3/7 activities (1 805.28±6.17, both P>0.05). The difference of S100 β protein expression was significant among the three groups ( F=490.80, P<0.01). Compared with the control group ((0.18±0.01)μg/L), the concentration of S100β protein in the surgery group ((2.13±0.02)μg/L) decreased ( P<0.01), while there was no significant difference in the surgery + nobiletin group ((0.16±0.01) μg/L, P>0.05). The expression levels of IBA-1 protein ( F=10.83) and TNF-α, IL-1, IL-1β and IL-6 ( F=996.20, 221.40, 73.02, 174.13) were significantly different among the three groups (all P<0.01). The expression level of the neuroglial marker IBA-1 in the surgery group(1.36±0.02) was significantly higher than that in the control group (1.00±0.01, P<0.01), while the surgery + nobiletin group (1.03±0.01) had no significant different compared with control group ( P>0.05). The levels of inflammatory factors, including TNF-α, IL-1, IL-1β and IL-6, in the brain of rats treated with nobiletin ((49.06±3.63)pg/mg, (2.09±0.43)pg/mg, (16.27±0.80)pg/mg, (2.11±0.19)pg/mg) were significantly lower than those in the surgery group((145.10±6.46)pg/mg, (5.67±0.43)pg/mg, (27.88±3.43)pg/mg, (4.74±0.32)pg/mg, all P<0.01). Conclusion:Nobiletin can obviously alleviate POCD symptoms caused by sevoflurane inhalation anesthesia.

Objective To explore the anti-tumor effect and the influence of antitumor immunity of PD-L1/PD-1 blocked by PD-1 antibody combined with cisplatin. Methods Tumor models were established by injecting TC-1 cells into C57BL/6 mice, and the mice were divided into four groups (n = 4). The tumor growth curves and survival curves were drawn to observe the anti-tumor effect. The tumors were then removed; and the PD-L1 and CD8+ T cells were analyzed by immunohistochemical method. Results The anti-tumor effect was greater in the cisplatin group , PD-1 antibody group , and PD-1 antibody plus cisplatin group than in the control group (P < 0.05). Expression of PD-L1 in the tumor tissues was markedly increased in the cisplatin group and it was obviously decreased in the combination group (P < 0.05). CD8+ T cells decreased in the cisplatin group; and expression of CD8+ T cells was significantly increased the combination group (P < 0.05). Conclusion The anti-tumor effect and anti-tumor immunity of cisplatin are enhanced by blocking PD-L1/PD-1 pathway with PD-1 antibody.

Objective To investigate the effects of chronic ethanol exposure and withdrawal on the expression of actin-binding protein cofilin,p-cofilin and cyclin-dependent kinase-5 (cdk5) in the nucleus accumbens and striatum in rat brain.Methods Twenty-four male SD rats were randomly divided into one control group and three experimental groups.In the experimental groups,ethanol was administered in drinking water at the concentration of 6％ (V/V) for two months.Rats in control group drank normal drinking water.After two months ethanol was removed and ethanol withdrawal syndromes were evaluated.Rats were sacrificed on withdrawal 0 h,withdrawal 6 h and withdrawal 2 d.The expression levels of cofilin,p-cofilin(ser3)and cdk5 in the rat brain were measured by immunohistochemistry methods.Results Withdrawal syndrome scores of ethanol fed rats were obviously higher than those of control rats after ethanol was removed,the highest score occurred at 6 h after ethanol withdrawal.In the nucleus accumbens area of rat brain,the levels of cofilin on withdrawal 0 h significantly decreased compared with control group ((0.31±0.05),(0.39± 0.05),P＜ 0.05).The levels of cdk5 on withdrawal 0 h and withdrawal 6 h significantly increased compared with control group((0.36±0.07),(0.34±0.07),(0.25±0.05),P＜0.05).In the striatum of rat brain,the levels of cofilin on withdrawal 0 h significantly decreased compared with control group ((0.26±0.04),(0.34±0.05),P＜0.05).The levels of p-cofilin on withdrawal 6 h significantly increased compared with control group((0.43±0.06),(0.30±0.06),P＜0.01).The levels of cdk5 on withdrawal 0 h significantly increased compared with control group((0.35±0.06),(0.26±0.05),P＜0.05),and the levels of cofilin on withdrawal 6 h significantly decreased compared with control group((0.37±0.06),(0.26±0.05),P＜0.01).Conclusion Chronic ethanol exposure can induce the development of ethanol dependence,and it accompanies with changes in the expression of actin-binding protein and cdk5 in the brain of rats.

A 87-day-old female patient presented with patchy erythema on the right body and ipsilateral limb deformity after birth, and visited the hospital in January 2012. Skin examination showed obvious red plaques on the right lower abdomen, right buttock, right perineum, right leg and right foot, with yellow scales on the surface and clear boundaries, and there was no obvious exudation or odor; pale yellow verrucous hyperplasia was observed on the right lower jaw, right neck, right axilla, and on the back of the first to fourth fingers of the right hand; only 1 interphalangeal joint and no nail plate was observed on the second, third and fourth toes of the right foot. The X-ray of the right foot showed deformity and bone defects. The patient was diagnosed with CHILD syndrome, and treated with intermittent oral acitretin capsules, and topical vaseline ointment, tacalcitol ointment and weak-potency glucocorticoid ointment for more than 6 years. The skin lesions regressed during treatment, but occurred repeatedly after withdrawal. Thereafter, the patient was switched to simvastatin ointment for 2 years. The skin lesions increased proportionally with the increase of age, and the affected limb grew a little more rapidly than the healthy limb. By January 2020, the skin lesions on the right lower jaw, right neck, and the first to fourth fingers of the right hand had subsided, but new band-like hypertrophic verrucous plaques occurred on the fifth finger of the right hand; the skin lesions on the right leg were slightly improved, and no involvement of other systems was observed.

Objective: To analyze the clinicopathologic characteristics of poorly-differentiated chordoma with INI1 loss in children and to discuss the differential diagnosis. Methods: The clinical, radiological, histopathological profiles and molecular pathologic characteristics of two pediatric poorly differentiated chordoma cases with INI1 loss were reviewed. Results: The patients were a girl and a boy. Both lesions involved the slope. Both patients were presented with progressive muscle weakness or neck pain. Radiological examination showed clivus bone destruction and compression of the brain stem and cervical spinal cord. Histologically, the tumor cells lacked typical organization and were associated with inflammatory cells infiltration. On high power field, the tumor cells were ovoid or fusiform with prominent atypia, vacuolated nuclei and prominent nucleoli. By immunohistochemistry, the tumor cells expressed cytokeratin, epithelial membrane antigen, brachyury and were negative for INI1. In both cases, INI1 gene deletion was detected by FISH. Conclusions Poorly-differentiated chordoma with INI1 loss mainly occurs in children. The morphology is different from classical chordoma.INI1 gene deletion is detectable by FISH. It can be distinguished from atypical teratoid/rhabdoid tumors and other neoplasms by the identification of nuclear brachyury expression. The loss of INI1 expression in poorly-differentiated chordoma might be associated with a poorly-differentiated morphology and an adverse prognosis.

OBJECTIVE: To determine the effect of preamputation pain on the behavioral response and astrocytic activation in the spinal cord of amputated rats, and to assess the association between preamputation pain and chronic amputation-related pain. METHODS: A total of 84 adult male SD rats were randomly distributed into an NA group (n=42) and a PA group (n=42). The NA group was intraplantarly injected with saline 100 μL, while the PA group was intraplantarly injected with complete Freund's adjuvant (CFA) 100 μL in both cases at 7 d before the amputation. Thermal withdrawal latency (TWL) was measured before the injection and at 1, 3, 5, and 7 d after the injection. All rats were amputated on the 7th day. The TWL, diet and water intake were measured on 1, 3, 5, 7, 10, 14, 17, 21, and 28 d after the amputation. Expression of glial fibrillary acidic protein (GFAP) in the L4-6 of spinal cord was measured by immunohistochemistry before the saline/ CFA injection, 7 d after the injection and 1, 3, 5, 7, 10 d after the amputation.. RESULTS: The TWL significantly decreased on 1, 3, 5, and 7 d after the intraplantar administration of CFA compared with the basic value in the PA group (P<0.05), while there was no difference between 1, 3, 5, and 7 d after the intraplantar administration of saline and the basic value in the NA group (P>0.05). In addtions to the basic value, the TWL of the PA group was shorter than that of the NA group at the above-mentioned time-points (P<0.05). Compared with the preoperative level, the diet and water intake decreased significantly after the amputation in both groups, but recovered to the preoperative levels, by 3 d after the amputation in the NA group, and by 5 d after the amputation in the PA group. Compared with the TWL of the residual limb on the day of amputation, the TWL of the residual limb increased significantly 3 d after the amputation and remained elevated until 28 d after the amputation in the NA group (P<0.05), while there was no difference between each time point after the amputation and the day of the amputation in the PA group. Compared with the basic value, there was an obviously high expression of GFAP in the NA group beginning on the day of amputation and in the PA group 7 d after the CFA injection (P<0.05). After the amputation, the expression of GFAP was significantly higher in the PA group (P<0.05). CONCLUSION Preamputation pain delays the recovery and activates the spinal astrocytes which may turn the acute postoperative pain into a chronic one.
Amputation ; Animals ; Astrocytes ; physiology ; Behavior, Animal ; Male ; Pain ; physiopathology ; Pain, Postoperative ; physiopathology ; Preoperative Period ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; physiopathology

OBJECTIVE: To investigate the effect of pre-amputation pain block on the N-methyl-D-aspartate receptor activation in the central nervous system of amputated rats, and the association between pre-amputation pain block and chronic amputation-related pain. METHODS: Thirty-six adult male SD rats were randomly assigned to an NA group (n=12), a PA group (n=12) and a PAB group (n=12). Group NA was intraplantarly injected saline l00 μL while group PA and group PAB were intraplantarly injected complete Freund adjuvant (CFA) 100 μL. The sciatic nerve of group NA and group PA were freed from surrounding tissue, and that of group PAB was blocked by bupivacaine under pentobarbital sodium anesthesia 5 days after the injection. Thermal withdrawal latency (TWL) was measured before and after the injection. All rats were amputated at the scheduled survival time. The expression of N-methyl-D-aspartate receptor (NR2B) was measured by immunohistochemistry in L4-6 of the spinal cord and the anterior cingulated cortex 7 days after the amputation procedure. RESULTS: The TWL after intraplantar administration of CFA in group PA and group PAB decreased significantly compared with the baseline value (P<0.05), while the saline treated control group remained unchanged. Besides the basic value, the TWL of group PA was shorter than that of group NA at the above-mentioned time-points (P<0.05). Compared with the basic value and group NA, the TWL of group PAB after the block increased significantly (P<0.05). Compared with group NA and group PAB, group PA had a remarkably high expression of NR2B (P<0.05), while there was no difference between group PA and group PAB. CONCLUSION Pre-amputation pain may activate N-methyl-D-aspartate receptor of the central nervous system, which may lead acute postoperative pain to chronic pain. It is necessary to treat pre-amputation pain.
Amputation ; Animals ; Bupivacaine ; administration & dosage ; Freund's Adjuvant ; administration & dosage ; Gyrus Cinguli ; metabolism ; Male ; Nerve Block ; Pain Measurement ; Pain, Postoperative ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Sciatic Nerve ; Spinal Cord ; metabolism

Objective: To evaluate the diagnostic value of a histologic scoring system in congenital biliary atresia and its prognostic relevance. Methods: From January 2017 to June 2018 at Children′s Hospital of Fudan University, 172 wedge liver biopsy specimens were obtained from infants with neonatal cholestasis [119 patients with congenital biliary atresia (CBA) and 53 patients with non-obstructive cholestasis as control]. A pathologist, single-blinded to the final diagnosis, made the histological diagnosis individually based on an 8-feature (portal ductal proliferation, bile duct reaction, bile plugs in portal ductules, liver fibrosis, edema in portal region, cholestasis, inflammatory cells infiltration in portal region, and ductal plate malformation), 21-point scoring system. Results: The main pathologic changes of biliary atresia were hepatocyte cholestasis, hyperplasia of bile ducts, fibrosis and infiltration of inflammatory cells in the portal area. There were significant difference in the degree of portal edema, bile duct hyperplasia and fibrosis between two groups (P<0.01). In addition, there were characteristic bile duct thrombosis in 97.5%(116/119) of the cases and abnormal development of bile duct plate in 9.2%(11/119) of the cases. Compared with non-CBA infant cholestasis group, the difference was statistically significant (P<0.05). The scoring system has high sensitivity, specificity (both 94.1%) and accuracy (94.3%) in the diagnosis of CBA. A score equal to or more than 11 points supported a diagnosis of CBA; whereas a score less than 11 points might suggest cholestasis. The degree of hepatic fibrosis and ductal plate malformation were related to prognosis. Conclusions The liver pathology scoring system (8-feature, 21-point) is more accurate in diagnosing CBA than previous methods, which may guide the clinicopathological diagnosis. This histological scoring system also helps to assess the prognosis of CBA.

Objective To investigate the mechanisms of miRNA-15a and miRNA-16 in the process of reversing cisplatin resistance in ovarian cancer.Methods Human ovarian cancer cisplatin-resistant cell lines CoC1/DDP were transfected with miRNA-15a and miRNA-16 mimics and treated with cisplatin.qRT-PCR was used to detect the expression levels of miRNA-15a and miRNA-16 in normal CoC1/DDP cell group,cisplatin treated group,negative control group,miRNA-15a transfected group,miRNA-16 transfected group and overexpressed Bmi-1 plasmid.Western blot was used to detect the expression level of Bmi-1 in each group,CCK-8 and Annexin V/PI were used to detect cell survival and apoptosis,and γ-H2AX immunofluorescence was used to detect cell apoptosis.Results The CoC1/DDP ovarian cancer cell line shows low expression of miRNA-15a and miRNA-16,and high expression of Bmi-1 protein,which makes it resistant to cisplatin.When the levels of miRNA-15a and miRNA-16 are overexpressed,the Bmi-1 protein decreases(P<0.05),leading to a decrease in cell survival rate(P<0.05),a significant increase in DNA apoptosis(P<0.05),and more severe DNA damage(P<0.05).Overexpression of Bmi-1 plasmid can increase cell viability(P<0.05)and reduce the rate of cell apoptosis(P<0.05).Conclusion The Bmi-1 protein may be a target for the regulation of miRNA-15a and miRNA-16,and overexpression of miRNA-15a and miRNA-16 can increase the sensitivity of ovarian cancer cells to cisplatin by reducing the Bmi-1 protein.This provides a new idea for predicting molecular markers of cisplatin resistance in ovarian cancer and overcoming drug resistance targets in ovarian cancer.

Objective To investigate the effect of esmolol pretreatment on Toll like receptor-4 (TLR4)/nuclear factor-kappa-B (NF-кB) pathway in rats with repeated cerebral ischemia/reperfusion (IR) injury. Methods Forty-eight adult male Sprague-Dawley rats were randomly allocated into sham-operated group, IR group and esmolol group (n=16). Rats in the IR group and esmolol group were injected intravenously with esmolol at a dose of 200 g/(kg?min) or normal saline for one h before surgery, and then, bilateral common carotid arteries were clipped to establish the repeated IR injury models. Bilateral common carotid arteries in rats of sham-operated group were only isolated but not clipped, and injected intravenously with normovolemic normal saline for one h. Learning and memory abilities of rats were measured by Morris water maze test before, and one, 3 and 7 d after surgery. Rats were euthanized and hippocampus tissues were excised. The wet to dry (W/D) ratio of the hippocampus was tested. The permeability of blood-brain barrier was determined by Evans blue (EB) method. The levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6 and IL-1β in the hippocampus were tested by enzyme linked immunosorbent assay (ELISA). The NF-кB p65 and TLR4 mRNA expressions in the hippocampus were determined by reverse transcription-polymerase chain reaction (RT-PCR). The NF-кB p65 and TLR4 protein expressions in hippocampus were detected by Western blotting. Results As compared with those in the sham-operated group, the escape latency was significantly prolonged and swimming distance was signficantly longer in rats of IR group one, 3 and 7 d after surgery (P<0.05), the W/D ratio of the hippocampus and the content of EB in brain tissues were significantly increased in the IR group (P<0.05), the levels of TNF-α, IL-6 and IL-1β in hippocampus were significantly increased in the IR group (P<0.05), and the NF-кB p65 or TLR4 mRNA and protein expressions in the hippocampus of IR group were statistically higher (P<0.05). As compared with IR group, the escape latency and swimming distance of rats in the esmolol group were significantly shortened one, 3 and 7 d after surgery (P<0.05), the W/D ratio of the hippocampus and content of EB in brain tissues of esmolol group were significantly decreased (P<0.05), the levels of TNF-α, IL-6 and IL-1β in the hippocampus of esmolol group were signficantly lower(P<0.05), and the NF-кB p65 or TLR4 mRNA and protein expressions in hippocampus of esmolol group were statistically lower in the esmolol group (P<0.05). Conclusion Esmolol preconditioning can alleviate cerebral injury and improve learning and memory abilities of rats with repeated cerebral IR injury, which may be involved in alleviating inflammation and suppressing TLR4/NF-кB pathway.