Objective To investigate the role of membrane-associated tyrosine/threonine-protein ki-nase 1(PKMYT1)in glucocorticoid(GC)-induced osteoblast(OB)apoptosis,providing a theoretical basis and potential therapeutic targets for early-stage steroid-induced avascular necrosis of the femoral head(SANFH).Methods(1)Mouse calvarial osteoblastic cells(MC3T3-E1)were selected for the study.The control group was cultured in standard medium,while the experimental group was subject-ed to osteogenic induction culture,with osteogenic capacity verified by alkaline phosphatase(ALP)and Alizarin Red S(ARS)staining.Then,mouse osteoblasts(mOB)were treated with different con-centrations of GC.After that,apoptosis was detected by using Annexin V-FITC/PI double staining as-say,while cell proliferation was assessed by using Cell Counting Kit-8(CCK8).Moreover,the expres-sions of anti-apoptotic protein B-cell lymphoma/leukemia-2(BCL-2),pro-apoptotic proteins cleaved caspase-3andcleavedcaspase-9(cleavedcaspase 3/9)weredetectedbyusing Westernblotting(WB).Meanwhile,proteomic analysis was employed to identify molecules potentially regulating GC-in-duced apoptosis in mOBs.What's more,quantitative real-time PCR(qPCR)and WB were used to further analyze PKMYT1 expression.(2)mOBs were treated with PKMYT1 inhibitor GSK-1520489A of different concentrations to screen the optimal one,and all subjects were then further divided into a control,a GC,a GSK-1520489A,and a GC+GSK-1520489A group.Later,the expression of PK-MYT1 and apoptosis-related proteins BCL-2 and cleaved caspase 3/9 of all groups were detected us-ing WB,and cell viability and cytotoxicity were evaluated by CCK8 assay,with cell proliferation by using 5-ethynyl-2'-deoxyuridine(EDU)assay and apoptosis by cell live/dead staining and Annexin V-FITC/PI double staining.(3)mOBs were infected with PKMYT1 overexpression lentiviral vectors,and its efficiency was verified by using immunofluorescence,qPCR,and WB.After successful overexpres-sion of PKMYT1,all cells were divided into the control,GC,PKMYT1 overexpression(OE),and OE+GC groups,whose cell proliferation was detected by EDU assay,and apoptosis was assessed by Annexin V-FITC/PI double staining and cell live/dead staining.(4)To verify the changes in PKMYT1 expression in human osteoblasts(hOB),hOBs extracted from human femoral heads of healthy individu-als were chosen into the control group,while those from patients with hormone-induced avascular ne-crosis of the femoral head(hSANFH)were selected into the hSANFH group.Then,PKMYT1 expres-sion in both groups was detected by using qPCR and WB.Results(1)After inducing the differentia-tion of mouse calvarial osteoblastic cells(MC3T3-E1)into mature osteoblasts,under the action of GC,compared with the control group,with the increase of GC concentration,the experimental group showed increased mOB apoptosis(P<0.01)and expression of cleaved caspase 3/9(P<0.01),but de-creased cell viability(P<0.01)and expressionof apoptosis-relatedprotein BCL-2(P<0.01).More-over,according to the proteomic sequencing,significant decrease was observed in the PKMYT1 expres-sion in mature mOBs treated with GC.(2)As to treatment of mOBs with different concentrations of PKMYT1 inhibitor GSK-1520489A,with the increase of concentration,cell viability decreased and cy-totoxicity increased(P<0.001).Moreover,compared with the control group,mOBs proliferation de-creased(P<0.001)and apoptosis increased(P<0.001)in the GSK-1520489A group.Meanwhile,com-pared with the GC group,mOB proliferation decreased(P<0.05)and apoptosis increased significantly(P<0.01)in the GC+GSK-1520489A group.(3)After overexpression of PKMYT1,in comparison with the control group,mOB proliferation increased(P<0.001)but apoptosis did not increase significantly(P>0.05)in the OE group.Moreover,compared with the GC group,mOB proliferation increased(P<0.001)but apoptosis decreased(P<0.001)significantly in the OE+GC group.(4)In hOBs extracted from human femoral head tissues,qPCR and WB results showed that PKMYT1 expression of the hSANFH group was significantly lower than the control group(P<0.001).Conclusion Down regulation of PKMYT1 expression promotes GC-induced apoptosis of mOBs.Conversely,over expression of PK-MYT1 inhibits GC-induced apoptosis of mOBs.Therefore,PKMYT1 may serve as a potential target for the early treatment of SANFH.

Objective To investigate the clinical efficacy of Shangke Huangshui medicated gauze in the treatment of small-area deep second-degree burn wounds with fire-heat injuring fluid type.Methods Sixty patients who were diagnosed as small-area deep second-degree burn wounds of fire-heat injuring fluid type in Foshan Hospital of Traditional Chinese Medicine from January 2024 to July 2024,were selected as the research objects.The patients were randomly divided into trial group and control group by random number table method,with 30 cases in each group.The trial group was treated with external application of Shangke Huangshui medicated gauze,and the control group was treated with external application of Silver Sulfadiazine Cream.The treatment lasted for 21 days,and then the patients were followed up for 7 days.The changes of Visual Analogue Scale(VAS)score of wound pain,and serum levels of C-reactive protein(CRP),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the two groups were observed before and after treatment.The wound healing rate,wound healing time,bacterial infection of the wound,and adverse reactions were compared between the two groups.Results(1)During the treatment,there was no fell-off case in the trial group while there was one patient falling off from the control group.Eventually,a total of 59 patients were included in the statistical analysis,including 30 in the trial group and 29 in the control group.(2)On day 14 and 21 of treatment,the wound healing rates of the two groups were significantly higher than those on day 7 of treatment(P＜0.05),and wound healing rates in the trial group on the day 14 and 21 of treatment were significantly superior to those of the control group(P＜0.05).(3)The time for the complete healing of wound in the trial group was(22.07±2.30)days,which was significantly shorter than that of the control group[(27.07±4.10)days],and the difference was statistically significant(P＜0.05).(4)After 7,14 and 21 days of treatment,the VAS scores of wound pain in the two groups were lowered compared with those before treatment(P＜0.05),and the VAS scores in the trial group were significantly lower than those in the control group(P＜0.05).(5)On day 7 of treatment,the levels of serum CRP,IL-6 and TNF-α in the two groups were lowered compared with those before treatment(P＜0.05),and the levels in the trial group were significantly lower than those in the control group(P＜0.05).(6)On day 7 of treatment,the positive rate of bacterial culture for wound discharge in the trial group was 6.67%(2/30),which was significantly lower than 27.59%(8/29)in the control group,and the difference was statistically significant(P＜0.05).(7)There were no serious adverse events or adverse reactions occurring in the two groups during the trial.Conclusion Shangke Huangshui medicated gauze can accelerate the healing of burn wounds,shorten the wound healing time,reduce the wound infection rate and the level of serum inflammatory factors,and has fewer adverse reactions with high safety.

Objective To investigate the role of membrane-associated tyrosine/threonine-protein ki-nase 1(PKMYT1)in glucocorticoid(GC)-induced osteoblast(OB)apoptosis,providing a theoretical basis and potential therapeutic targets for early-stage steroid-induced avascular necrosis of the femoral head(SANFH).Methods(1)Mouse calvarial osteoblastic cells(MC3T3-E1)were selected for the study.The control group was cultured in standard medium,while the experimental group was subject-ed to osteogenic induction culture,with osteogenic capacity verified by alkaline phosphatase(ALP)and Alizarin Red S(ARS)staining.Then,mouse osteoblasts(mOB)were treated with different con-centrations of GC.After that,apoptosis was detected by using Annexin V-FITC/PI double staining as-say,while cell proliferation was assessed by using Cell Counting Kit-8(CCK8).Moreover,the expres-sions of anti-apoptotic protein B-cell lymphoma/leukemia-2(BCL-2),pro-apoptotic proteins cleaved caspase-3andcleavedcaspase-9(cleavedcaspase 3/9)weredetectedbyusing Westernblotting(WB).Meanwhile,proteomic analysis was employed to identify molecules potentially regulating GC-in-duced apoptosis in mOBs.What's more,quantitative real-time PCR(qPCR)and WB were used to further analyze PKMYT1 expression.(2)mOBs were treated with PKMYT1 inhibitor GSK-1520489A of different concentrations to screen the optimal one,and all subjects were then further divided into a control,a GC,a GSK-1520489A,and a GC+GSK-1520489A group.Later,the expression of PK-MYT1 and apoptosis-related proteins BCL-2 and cleaved caspase 3/9 of all groups were detected us-ing WB,and cell viability and cytotoxicity were evaluated by CCK8 assay,with cell proliferation by using 5-ethynyl-2'-deoxyuridine(EDU)assay and apoptosis by cell live/dead staining and Annexin V-FITC/PI double staining.(3)mOBs were infected with PKMYT1 overexpression lentiviral vectors,and its efficiency was verified by using immunofluorescence,qPCR,and WB.After successful overexpres-sion of PKMYT1,all cells were divided into the control,GC,PKMYT1 overexpression(OE),and OE+GC groups,whose cell proliferation was detected by EDU assay,and apoptosis was assessed by Annexin V-FITC/PI double staining and cell live/dead staining.(4)To verify the changes in PKMYT1 expression in human osteoblasts(hOB),hOBs extracted from human femoral heads of healthy individu-als were chosen into the control group,while those from patients with hormone-induced avascular ne-crosis of the femoral head(hSANFH)were selected into the hSANFH group.Then,PKMYT1 expres-sion in both groups was detected by using qPCR and WB.Results(1)After inducing the differentia-tion of mouse calvarial osteoblastic cells(MC3T3-E1)into mature osteoblasts,under the action of GC,compared with the control group,with the increase of GC concentration,the experimental group showed increased mOB apoptosis(P<0.01)and expression of cleaved caspase 3/9(P<0.01),but de-creased cell viability(P<0.01)and expressionof apoptosis-relatedprotein BCL-2(P<0.01).More-over,according to the proteomic sequencing,significant decrease was observed in the PKMYT1 expres-sion in mature mOBs treated with GC.(2)As to treatment of mOBs with different concentrations of PKMYT1 inhibitor GSK-1520489A,with the increase of concentration,cell viability decreased and cy-totoxicity increased(P<0.001).Moreover,compared with the control group,mOBs proliferation de-creased(P<0.001)and apoptosis increased(P<0.001)in the GSK-1520489A group.Meanwhile,com-pared with the GC group,mOB proliferation decreased(P<0.05)and apoptosis increased significantly(P<0.01)in the GC+GSK-1520489A group.(3)After overexpression of PKMYT1,in comparison with the control group,mOB proliferation increased(P<0.001)but apoptosis did not increase significantly(P>0.05)in the OE group.Moreover,compared with the GC group,mOB proliferation increased(P<0.001)but apoptosis decreased(P<0.001)significantly in the OE+GC group.(4)In hOBs extracted from human femoral head tissues,qPCR and WB results showed that PKMYT1 expression of the hSANFH group was significantly lower than the control group(P<0.001).Conclusion Down regulation of PKMYT1 expression promotes GC-induced apoptosis of mOBs.Conversely,over expression of PK-MYT1 inhibits GC-induced apoptosis of mOBs.Therefore,PKMYT1 may serve as a potential target for the early treatment of SANFH.

Memantine is a non-competitive N-methyl-D-aspartate receptor (NMDAR) antagonist clinically approved for moderate-to-severe Alzheimer's disease (AD) to improve cognitive functions. There is no report about the proteomic alterations induced by memantine in AD mouse model yet. In this study, we investigated the protein profiles in the hippocampus and the cerebral cortex of AD-related transgenic mouse model (3×Tg-AD) treated with memantine. Mice (8-month) were treated with memantine (5 mg/kg/bid) for 4 months followed by behavioral and molecular evaluation. Using step-down passive avoidance (SDA) test, novel object recognition (NOR) test and Morris water maze (MWM) test, it was observed that memantine significantly improved learning and memory retention in 3xTg-AD mice. By using quantitative proteomic analysis, 3301 and 3140 proteins in the hippocampus and the cerebral cortex respectively were identified to be associated with AD abnormalities. In the hippocampus, memantine significantly altered the expression levels of 233 proteins, among which PCNT, ATAXIN2, TNIK, and NOL3 were up-regulated, and FLNA, MARK 2 and BRAF were down-regulated. In the cerebral cortex, memantine significantly altered the expression levels of 342 proteins, among which PCNT, PMPCB, CRK, and MBP were up-regulated, and DNM2, BRAF, TAGLN 2 and FRY1 were down-regulated. Further analysis with bioinformatics showed that memantine modulated biological pathways associated with cytoskeleton and ErbB signaling in the hippocampus, and modulated biological pathways associated with axon guidance, ribosome, cytoskeleton, calcium and MAPK signaling in the cerebral cortex. Our data indicate that memantine induces higher levels of proteomic alterations in the cerebral cortex than in the hippocampus, suggesting memantine affects various brain regions in different manners. Our study provides a novel view on the complexity of protein responses induced by memantine in the brain of AD.
Alzheimer Disease ; Animals ; Axons ; Brain ; Calcium ; Cerebral Cortex ; Cognition ; Computational Biology ; Cytoskeleton ; Hippocampus ; Learning ; Memantine ; Memory ; Mice ; Mice, Transgenic ; N-Methylaspartate ; Proteome ; Ribosomes ; Water

Early portal vein thrombosis is a rare but serious complication after liver transplantation,also is one of the main causes which lead to graft loss and receptor death. We collected domestic and foreign relevant data,and summarized and discussed the causes, clinical manifestation, imaging diagnosis of early portal vein thrombosis after liver transplantation. Early portal vein thrombosis after liver transplantation caused by multiple risk factors synergy. It was reported incidence of 1% to 2%. Clinical manifestations were concealment and lacked of specificity. Abdominal ultrasound and computed tomography angiography ( CTA) imaging methods of combi?ning could help early clinical findings,When necessary magnetic resonance angiography( MRA) . If each check negative,portal vein angiography could make a definitive diagnosis. Intravascular interventional therapy with small trauma, less complications and high success rate for advantages gradually became first?line treat?ments. Surgical treatment is not only as traditional effective treatments,but also an effective remedial measures after interventional treatment failure.

Artemisinin is the antidote to malaria, and has made hundreds of millions patients get rid of the disease since application in clinic. At present, the main accesses to artemisinin are directly extracted from Artemisia annua,chemical synthesis and biosynthesis. By comparing the differences of artemisinin production process between China and abroad, this research analyzes the reasons of the problem, and put forward their views and suggestions, so as to provide references for Artemisia annua further research.

Adult hepatoblastoma is extremely rare in clinical practice, and its pathogenesis is unknown. At present, there is a lack of understanding of this disease, and therefore clinical misdiagnosis is common. This review introduces the pathogenesis, clinical manifestations, and radiological and pathological characteristics of hepatoblastoma, focuses on the choice of treatment for adult hepatoblastoma, and explains briefly the latest research progress in biological therapy. Comparison of different treatments suggests that comprehensive treatment based on radical resection is still the best choice. However, the curative effect is poor since the disease is highly malignant and its treatment experience is insufficient. The newly reported target for biological therapy may represent a breakthrough for the treatment of the disease, but its clinical efficacy remains to be evaluated. The pathogenesis of the disease in adults and children has not been confirmed as the same, and the review indicates that further clinical research is still needed.

OBJECTIVE:To establish the fingerprint of Artemisia apiacea to identify common model and evaluate the quality coherence of the commercially available A. apiacea decoration pieces in Chongqing. METHODS:HPLC method was performed on the column of Waters XTerra C18 with the mobile phase consisted of methanol-0.1% phosphoric acid(gradient elution)at the flow rate of 1.0 ml/min. The detection wavelength was 220 nm,column temperature was 30 ℃ and the sample size was 10 μl. The simi-larity of 12 batches of samples was evaluated by TCM Chromatogram Fingerprint Similarity Evaluation System(2004A edition);a common model of fingerprints was identified by principal component analysis(PCA)and cluster analysis(CA). RESULTS:Total-ly 11 batches of samples were screened to establish common model of fingerprints and 16 common peaks were obtained,among which,3 common components were identified;RSDs of precision,stability and reproducibility tests were lower than 2.53%. The similarity of 11 batches of A. apiacea and reference chromatogram was more than 0.990. CONCLUSIONS:The method is stable and easy,and can provide reference for the scientific quality evaluation and control of A. apiacea;the quality of commercially available A. apiacea decoration pieces is generally good,however,there is still a little number of batches with poor quality. The production procedures and supervision of A. apiacea decoration pieces should be strengthened.

BACKGROUND: Pancreatic stem cells can be differentiated into endocrine functioning cells under a suitable cultivation condition, suggesting that pancreatic stem cells can be used as a new source of islet in treating diabetes mellitus.OBJECTIVE: To discuss the expression of stem cell in different purities of pancreatic islets.DESING: A controlled observation based on cells.SETTING: Third People's Hospital of Mianyang City.MATERIALS: The experiment was conducted in the Center Laboratory (Key Laboratory of Chongqing City), the First Affiliated Hospital of Chongqing Medical University from March 2005 to March 2006. Thirty healthy male SD rats, of clean grade, aged 50 days were used in this study. During the experiment, the disposal of animals corresponded to Animal Ethical Standard. Cytokeratin (CK)-19 and β-actin were designed by Shanghai Invitrogen Biological Co., Ltd.Brdu reagent, mouse anti-rat Brdu monoclonal antibody, rabbit anti-mouse (RAM) CK-19 polyclonal antibody and Brdu immunohistochemical kit were from Boster Co., Ltd. (Wuhan); HistostainTM -DS double immunohistochemical staining kit was purchased from Beijing Zhongshan Company.METHODS: Rats were intraperitoneally injected with Brdu for labeling, and subjected to perfusion with Ⅴ-type collagenase via pancreatic ducts, with pancreas being excised, ripped, beaten up, digested and centrifuged to obtain pancreatic islet sediments. After being precipitated, the sediments were divided into 3 groups. Group A: The isolated sediments of islets were not purified; Group B: The sediments were added slowly with 25% Ficoll-400 and Hanks solution for purification; Group C: The sediments were slowly added with 25% Ficoll-400, 11% Ficoll-400 for purification in order.MAIN OUTCOME MEASURES: Expressions of Brdu and CK-19-positive cells in the islet samples were detected by immunohistochemical method, and mRNA expression of CK-19 in the different purities of islets was detected by reverse transcription-polymerase chain reaction.RESULTS: ① The purity of islets in the group A was the lowest. The purity of islets obtained by different purification techniques showed significances among the three groups (F =89.42, P ＜ 0.05). ②Positive expressions of Brdu and CK-19 in the group A were significantly higher than those in the group B and group C (F =18.64, 22.12, 38.61. P ＜0.01). ③Expression of CK-19 mRNA in the group A was significantly higher than that in other groups, with group C showing the lowest.CONCLUSION: Expressions of Brdu and CK-19-positive cells exist in different purities of islets, suggesting the existence of more stem cells in the low purify of islets.

BACKGROUND: To synthetically analyze the correlation between cytokeratin 19 and pancreas stem cells.DATA SOURCES:References about the correlation between cytokeratin 19 and pancreas stem cells were retrieved in Medline and Ovid database with the key words of "cytokeratin, pancreas, stem cells, marker, differentiation" in English from January 2000 to May 2006.STUDY SELECTION:Literatures about the correlation between cytokeratin 19 and pancreas stem cells were retrieved and those characterized by strong pertinency, published in the near future and selected from authoritative journals were selected in this paper.DATA EXTRACTION: Among 86 articles, 35 of them met the inclusion criteria and 51 were excluded.DATA SYNTHESIS:Pancreas stem cells are characterized by multi-directional differentiation.Cytokeratin 19 has a positive expression in pancreas stem cells and a molecular sign of pancreas stem cells. This provides a new way to further understand bionomics of pancreas stem cells,study separation,purification,transconformation and direction differentiation to islet cells, and treat diabetes mellitus with transplantation of pancreas stem cells.CONCLUSION: Expression of cytokeratin 19 is observed during the differentiation of pancreas stem cells and cytokeratin19 is a sign of pancreas stem cells.