Objective To evaluate the effects of penehyclidine hydrochloride (PHC) on the expression of Toll like receptor-4 (TLR4) and nuclear factor kappa B (NF-κcB) in a rat model of sepsis-induced acute lung injury (ALI).Methods Seventy-two adult male Sprague-Dawley rats,weighing 180-220 g,were randomly divided into 3 groups (n =24 each) using a random number table:shame operation group (group S),ALI group and PHC group.Sepsis-induced ALl was induced by cecal ligation and puncture in anesthetized rats.In group PHC,PHC 0.45 mg/kg was intramuscularly injected immediately after cecal ligation and puncture.At 6,12,24 and 36 h after ligation,the broncho-alveolar lavage fluid (BALF) was collected for detection of TNF-α and IL-6 concentrations and the lungs were removed for determination of the expression of TLR4 (using RT-PCR and Western blot) and NF-κBp65 (using Western blot) in lung tissues and for microscopic examination.The pathological changes of lungs were scored.The wet to dry lung weight (W/D) ratio was calculated and the activity of myeloperoxidase (MPO) in lung tissues was measured.Results As compared with S group,the IL-6 concentrations in BALF at 6,12 and 24 h after ligation,TNF-α concentration in BALF at 6 and 12 h after ligation,and the expression of TLR4 and NF-κBp65,pathological scores,W/D ratio and MPO activity at each time point were significantly increased in ALI group (P ＜ 0.05).Compared with ALI group,the TNF-α concentration in BALF at 6 and 12 h after ligation,and IL-6 concentrations in BALF,the expression of TLR4 and NF-κBp65,pathological scores,W/D ratio and MPO activity at each time point were significantly decreased in group PHC (P ＜ 0.05).Conclusion PHC inhibits NF-κB activation and decreases inflammatory responses through blocking TLR4 expression in lung tissues,thus attenuating sepsis-induced ALI in rats.

Objective To explore the roles of glycogen synthase kinase-3β (GSK-3β) in cognitive dysfunction and emotional alterations after rat's chronic cerebral ischemia.Methods 51 SD rats were randomly divided into sham group (sham group,n=17),chronic cerebral ischemia group (2VO group,n=17),chronic cerebral ischemia group + LiCl group (2VO + LiCl group,n=17),according to the table of random number.All groups were intraperitoneally injected with LiCl or saline on 3rd,7th,14th,21 st and 28th day,and then produced the chronic cerebral ischemia models.On the 28th day after model,the spatial learning and memory,fear memory,and anxiety emotion were detected.Results The Morris water maze test showed that 2VO group spent longer latent time searching and finding the platform than sham group (4th day P＜0.01,3rd,5th,6th,7th day P＜0.05).2VO+LiCl group spent shorter latent time than 2VO group (4th day P＜0.01,5th,6th,7th day P＜0.05).After removing the platform,2VO group spent longer time arriving the former location than sham group (P＜0.05).And 2VO+LiCl group spent dramatically different time compared to 2VO group (P＜0.01).Step-down test showed 2VO group spent shorter latent time than sham group (2VO group:(41.00±1.87)s,sham group:(44.55±2.77)s) (P＜0.05).2VO+ LiCl group spent dramatically longer latent time compared to 2VO group (2VO +LiCl group:(43.40± 1.35)s) (P＜ 0.05).2VO group made much more mistakes times than sham group (P＜0.05).2VO+LiCl group made dramatically less mistakes times compared to 2VO group (P＜0.05).The elevated plus maze test showed 2VO group had much less ratio of retention time in open arms (among total arms retention time) than sham group (2VO group:(0.23± 0.01),sham group:(0.25± 0.01)) (P＜ 0.01).2VO + LiCl group had much larger ratio than 2VO group (2VO + LiCl group:(0.24±0.01),P＜0.05).2VO group had much less ratio of entry times in open arms (among total arms entry times) than sham group (P＜0.01).2VO+LiC1 group had much larger ratio than 2VO group(P＜0.05).Conclusion Chronic cerebral ischemia can lead to deterioration of learning and memory,and anxiety emotion for rats.However,inhibition of GSK-3β can ameliorate these alterations.

Objective To explore the effects of 4-phenylbutyric acid(PBA) on cognitive dysfunction after chronic cerebral hypoperfusion and underlying mechanisms.Methods 62 male Sprague Dawley rats in clean degree were divided into sham group(n=14),chronic cerebral ischemia group(bilateral carotid arteries occlusion,2VO group,n=24),and chronic cerebral ischemia with PBA treatment(2VO+PBA group,n=24).The chronic cerebral ischemia models were produced by the occlusion of bilateral common caroid artery for 1 month.During the hypoperfusion,the rats were injected intraperitoneally with 11.25 mg · ml-1 PBA(90 mg · kg-1 · d-1) or equal volume of saline once a day for 3 days followed by every other day for 27 days.Learning and memory abilities were tested by Morris water maze and novel object recognition test.The expression and distribution of NR2A in hippocampus were examined by Western blot and immunohistochemistry.Results Morris water maze test showed that the 2VO group had significantly longer latent time than sham group in searching platform (P＜0.01) (from 2nd day to 7th day latency time),and the 2VO+PBA group had dramatically shorter latent time than 2VO group (2nd,5th,6th,7th (P＜0.01),3rd(P＜0.05)).Then the rats' memory test showed that 2VO group spent markedly longer time than sham group to reach the location of the former platform(P＜0.01),but the 2VO+PBA group spent dramatically shorter latent time than 2VO group (P＜0.01).The novel object recognition test showed the exploration ratio and discrimination index of novel object in 2VO group were noticeably smaller than that in sham group (P＜0.01),but the exploration ratio and discrimination index of novel object in 2VO+PBA group were noticeably higher than that in 2VO group (P＜0.01).The Western blot data showed that the level of NR2A in 2VO group was significantly lower than that in sham group (P＜0.01),but the level of NR2A in 2VO+PBA group was significantly higher than that in 2VO group (P＜0.05).The level of NR2B in hippocampus of 2VO group had no significant difference with sham group and 2VO+PBA group (P＞0.05).Immunohistochemistry data was consistent with the data of Western blot for NR2A level and distribution.The ratio of p-CREB/total CREB in 2VO group(0.62±0.04) was remarkably lower than that in sham group(1.00±0.07),but the ratio of p-CREB/total CREB in 2VO+PBA group(0.97±0.07) was remarkably higher than that in 2VO group(P＜0.01).Conclusion NR2A reduction is associated with chronic cerebral hypoperfusion-induced cognitive impairment,which is rescued by the PBA treatment.It suggests that PBA may have a therapeutic effect on preventing chronic cerebral hypoperfusion-induced cognitive impairment.

Objective To explore the correlationship between Cystatin C and the cognitive dysfunction, encephalatrophy and the cerebral perfusion in Alzheimer's disease ( AD ) patients, and to analyze the probable mechanisms of the Cystatin C level alterations. Methods For the 61 male and 55 female AD patients,the levels of Cystatin C,Urea,Cr andβ2?microglobulin were detected by biochemistry analysis.The hypocampus volume and Ev?ans index were measured by MRI imaging. The cerebral perfusion was evaluated by magnetic resonance perfusion imaging of arterial spin labeling technique and image processing. The cognitive level of AD patients was done by MMSE scale and MoCA scale. Results The Cystatin C levels in serum of male AD patients were dramatically dif?ferent from female AD patients(male:(0.95±0.20)mg/L,female:(1.32±0.41)mg/L)(P=0.04). For male AD patients,age and Cystatin C had a correlationship among the hypocampus volume((6157.14±355.58)mm3),Evans index(0.33±0.02),MMSE score(21.66±7.97),and MoCA score(21.96±6.19)(age:P=0.040,0.049,0.035, 0.039;Cystatin C:P=0.035,0.038,0.037,0.035),but for female only age((79.52±8.82)years) had a correla?tionship with the hypocampus volume((6319.53±377.74)mm3),Evans index(0.31±0.02),MMSE score(22.93± 6.22),and MoCA score(23.41±8.20)(P=0.044,0.045,0.047,0.046). For male AD patients,the marked correla?tionship was showed between Cystatin C and the index of cerebral perfusion(P=0.034),but for female patients, there's no correlationship(P=0.086) . Conclusion The Cystatin C level is distinctly different in male and female AD patients. The Cystatin C level in male AD patients maybe be an index to predict the degree of the cognitive dys?function and the level of cerebral perfusion.

Objective To explore the roles and related mechanisms of Protein Phosphatase 2A(PP2A) in cognitive dysfunction after the chronic cerebral ischemia.Methods 70 male Sprague Dawley rats in clean degree were divided into sham group,chronic cerebral ischemia group (Bilateral carotid arteries occlusion,2VO),and chronic cerebral ischemia group with PP2A activation group(2VO+aPP2A).The rats were injected intraperitoneally with 1.88 μmol/ml sodium selenate(15 μmol · kg-1 · d-1) or equal volume of saline for 4 weeks.After one month,the chronic cerebral ischemia models were reproduced by the occlusion of bilateral common caroid artery.Then the abilities of learning and memory were tested by Morris water maze,electrophysiological indices were recorded to analyze the LTP changes,and destribution of synaptic vesicles was observed by electron microscope.Results Morris water maze test showed that the 2VO group had significantly longer latent time than sham group in searching platform(P＜0.05),and the 2VO+aPP2A group had dramatically shorter latent time (P＜0.01) than that of 2VO group.Then removing platform to test the rats memory,the data showed that 2VO group spent markedly longer time than sham group to reach the location of the former platform (sham group:(14.50±1.98)s ; 2VO group:(17.30±2.11) s) (P＜0.01),and the 2VO+aPP2A group((15.09± 1.45) s) spent dramatically shorter latent time(P＜0.05) than that of 2VO group.The electrophysiological data showed that 2VO group had the noticeably smaller field excitable postsynaptic potential slope (fEPSP) slope ratio between pre and post of the high frequency stimulations (Long-term potential,LTP) than sham group(sham group:1.69±0.27; 2VO group:2.02±0.137) (P＜0.01),and the 2VO+aPP2A group(1.86±0.19) had strikingly higher ratio than that of 2VO group(P＜0.01).The electromicroscope observation showed that presynaptic vesicles density of 2VO was remarkably lower than that of sham group (sham group:(4.51±0.29) /μm2 ; 2VO group:(2.02±0.14) /μm2) (P＜0.01),and presynaptic vesicles density of 2VO+aPP2A group((3.58±0.50) /μm2) was noticeably higher than that of 2VO group(P＜0.01).Conclusion Activating PP2A can prevent the cognitive dysfunction after chronic cerebral ischemia through regulating LTP and synaptic vesicle density.And PP2A is probably a potential target for preventing and treating the cognitive dysfunction after chronic cerebral ischemia.

Objective:To analyze clinical characteristics of ketosis-associated prurigo pigmentosa after ketogenic diet and bariatric surgery.Methods:Clinical data were collected from patients with ketosis-associated prurigo pigmentosa, who were diagnosed and treated in Department of Dermatology, Peking University People′s Hospital from September 2018 to September 2020. The clinical characteristics, sequelae and therapeutic effect of dietary modification were analyzed and summarized.Results:A total of 6 patients with ketosis-associated prurigo pigmentosa were collected, including 5 females who developed prurigo pigmentosa after ketogenic diet, and 1 male who developed prurigo pigmentosa after bariatric surgery. The skin lesions mainly involved the chest, back, waist and abdomen, and rarely involved the eyelids, axillae, elbows and mons pubis. Common skin lesions included urticaria-like erythema, papules and pigmentation arranged in a reticular distribution, and rare skin lesions included mung bean- to soybean-sized blisters, whose walls were liable to break. Among 5 patients undergoing routine urine analysis, 4 were positive (from + to ++++) for ketone bodies in the urine, and 3 were positive for urinary protein (+) . Pathological examination in 2 patients showed epidermal spongiosis, scattered necrotic keratinocytes, basal cell liquefaction, lymphocyte infiltration in the superficial dermis, and erythrocyte extravasation. The 6 patients were advised to eat staple foods. After dietary modification, 5 patients were nearly cured within 1 week; 1 patient, who continued ketogenic diet for weight loss, still received marked improvement after the treatment with minocycline at a dose of 100 mg/d in spite of restriction of carbohydrate intake. The levels of urinary ketone bodies and urinary protein in the 6 patients all returned to normal within 1 week after treatment.Conclusions:Ketosis plays an important role in the occurrence of prurigo pigmentosa. Dietary modification alone or adjuvant medical treatment such as minocycline is effective for the treatment of ketosis-related prurigo pigmentosa.

OBJECTIVE: To discuss the optimal operation mode for the management of donated drugs.METHODS: The possible problems existed in the management of donated medicines were found out through summarizing our practical experience in the inventory,storage and use etc of the donated drugs.RESULTS & CONCLUSIONS: In the management of donated medicines,emphasis should be attached to the inventory,storage and use of the donated medicines,meanwhile,a sound supervising system for the donation affairs should be set up to standardize the donation behavior,strengthen expiration date management and gradually establish the practical and effective work mode.

OBJECTIVE:To establish an assay method for Sustained-release palmatine tablet in beagle dog. METHODS:Sample separation was performed on Diamonsil ODS C18 column(250 mm?4.6 mm,5 ?m) with mobile phase consisted of acetonitrile-water (1∶1,containing NaH2PO4 3.4 g and SDS 1.7 g in every 1 000 mL water,pH was adjusted to 3 by H3PO4) with a flow rate of 1.0 mL?min-1.The column was kept at 35 ℃ and the detective wavelength was set at 342 nm.RESULTS:The linear serum concentration range was 0.005～0.5 ?g?mL-1(r=0.999 9) and the recovery rate was(102.22?2.48)%. CONCLUSION:The method was proved to be simple,accurate,sensitive and reproducible,and it was up to the standards for the study of pharmacokinetics and bioavailability of Sustained-release palmatine tablet in beagle dog.

OBJECTIVE:To study the adverse drug reactions(ADRs)induced by weight-reducing drugs and to provide references for rational administration in clinics.METHODS:Classification statistic and analysis were conducted on462cases with weight-reducing drug-induced ADRs reported in62kinds of domestic and overseas medicinal periodicals published during the period of1980～2004.RESULTS:With pseudo-5-HTA appetite-suppressing drugs showing the high incidence of ADRs,which stood at36.36%of the total.Multi-systems were involved in ADRs,with the cardiovascular system showing the highest proportion(53.68%).The prognostic mortality accounted for18.61%of the total.CONCLUSION:ADRs induced by weight-reducing drugs were associated with many factors,to avoid the occurrences of which,medical workers should give more directions and monitoring on administration of drugs.

OBJECTIVE To investigate the nuclear proteomes in mitochondrial DNA (mtDNA)-depleted A549 cells (Rho0 cells) and their parental cells (Rho+ cells),and to learn more about the nuclear responses to mitochondrial dysfunction.METHODS The nuclear proteomes of Rho and Rho + cells were characterized by two dimensional electrophoresis (2-DE) and SELDI-TOF ProteinChip technologies,the differentially expressed protein- spots were identified by MALDI-TOF mass spectrum (MS),the nucleophosmin and P53 expression were detected by Western blotting assay,and the mitochondrial memhrane potential (MMP) was measured by the laser scanning confocal microscope.RESULTS 2-DE results showed 11 protein-spots were down-regulated and 21 protein-spots were up-regulated in Rho0 cell nuclei.SELDI-TOF MS analysis with NP20 ProteinChips revealed 4 protein-peaks decreased in Rho0 cell nuclei.One down-regulated protein-spot was identified as elF-6,and 4 up-regulated proteinspots were identified as nucleophosmin,SFRS1,SFRS3 and hnRNP G,respectively.The increased expression of nucleophosmin in Rho0 cells was verified by Western blotting.Based on the clues from proteomic analysis,P53 expression in Rho0 cells was higher than in Rho + cells,and MMP was consistent in Rho + and Rho0 cells.CONCLUSION mtDNA-depletion induces nuclear proteome alteration.Rho0 cells can be used as a model to study the crosstalk between mitochondrion and nucleus.