Objective To investigate the effects of phosphocreatine postconditioning on cerebral ischemia-reperfusion(IR)injury in rats.Methods Thirty-six SD rats were randomly divided into three groups:groups Sham,IR (treated with normal saline)and PCr.IR was induced by intraluminal middle cerebral artery occlusion (MCAO).All treatments were given intravenously at the begining of reperfusion.Twenty-four hours after the reperfusion, neurological deficit score and magnetic resonance scan were performed.serum concentrations of malonaldehyde and 4-hydroxynonenal,cere-bral infarct volume and destruction of cerebral cortex were estimated.Neuronal apoptosis was further assessed by immunohistochemistry and immunofluorescent staining of caspase-3 and NeuN. Results Compared with group IR,phosphocreatine significantly decreased neurological deficit score, infarct volume,malonaldehyde and 4-hydroxynonenal levels(P < 0.05 ).Cortex structure was more complete,as well as neuronal apoptotic index was smaller in group PCr (P <0.05).Conclusion PCr can reduce cerebral infarct volume,thereby promote neurofunctional recovery.The mechanism of Pcr is related to reduced oxidative stress and inhibitted apopotosis during IR.

Objective To investigate protective effects of dexmedetomidine on oxygen-glucose deprivation/reperfusion(OGD/R)-induced neuronal apoptosis.Methods SH-SY5Y cells were differentiated to neurons with ATRA and followed by TPA.According to the results of preliminary experiment, OGD/R modle was constructed by oxygen-glucose deprivation(OGD) for 12 h and reperfusion(R) for another 12 h.During the start of the OGD, neurons were immediately divided into six groups: group D0(0 μmol/L dexmedetomidine), group D1(0.1 μmol/L dexmedetomidine), group D2 (1 μmol/L dexmedetomidine), group D3 (10 μmol/L dexmedetomidine), group D4(100 μmol/L dexmedetomidine), group D5 (1 000 μmol/L dexmedetomidine).After reperfusion 12 h, the cell viability was evaluated by the method of MTT.The cellular apoptosis was observed by flow cytometry method.The protective effects of different concentration dexmedetomidine on OGD/R-induced neuronal apoptosis were investigated.Then in chosen the exact group having protective effects, endoplasmic reticulum stress specific protein mesencephalic astrocyte-derived neurotrophic factor (MANF) and pro-apoptotic protein Caspase-3 and CHOP were detected by Westernblot method.Results Compared with group D0, there was no difference on the cell viability and cellular apoptosis induced by OGD/R in groups D1 and D2, but a significant decrease and increase in groups D4 and D5 (P<0.01 or P<0.05).And only group D3 had a neuroprotective effect, significantly increased the cell viability and inhibited the apoptosis (P<0.01).Further studys found that group D3 significantly up-regulated ER stress specific protein MANF (P<0.01) and inhibited up-regulation of Caspase-3 and CHOP (P<0.01).Conclusion These data suggest that 10 μmol/L dexmedetomidine had neuroprotective effect on OGD/R-induced neuronal apoptosis and significantly increased cell viability.Our results also indicate that up-regulation of ER stress specific protein MANF and inhibition of CHOP and Caspase-3 by MANF are involved in the neuroprotective effects of Dexmedetomidine.

Objective To investigate the expression and genetic alterations of KLF6 in hepatocellular carcinoma (HCC) and explore their functional mechanisms in the oncogenesis and development of HCC. Methods Real-time quantitative-PCR, direct sequencing and LOH approaches were used to detect KLF6 genetic abnormalities in HCC. The experiment had 2 groups, an experimental group and a control group. In the experimental group, the transfected plasmid pcDNA3.0 was recombined with KLF6 and tranfected into HCC HepG2 cells. MTT, flow cytometry and Western blotting were used to observe the effect of anti-oncogene wild type KLF6 on HepG2 cells by transgenic method for 48 h.Results Expression levels of KLF6 were significantly downregulated in HCCs(P＜0. 01), as detected by qRT-PCR. LOH occurred in 11 (52%) of the 21 tumors, and all the samples with LOH showed KLF6 down-regulation. The mutational frequency was 29%, and sequence changes located in activation domain of KLF6. Meanwhile, MTT assay showed a significant antiproliferative effect of the transfected wtKLF6 on HepG2 cells(42.7%, P＜0.05). Fluorescence-activated cell sorting analysis revealed that KLF6 induced apoptosis. Conclusion The deregulation of KLF6 together with genetic abnormalities of allelic imbalance and mutations may play an important role in HCC pathogenesis.

Objective To evaluate the left ventricular function and the synchrony of myocardial ischemic segments in patients with coronary heart disease (CHD) after PTCA and stent implantation by quantitative tissue velocity imaging (QTVI). Methods Thirty-six patients with isolated left anterior descending stenosis (≥75%) were examined by QTVI three days before, one week and one month after successful PTCA and stent implantation to measure the following items of 5 different left ventricular segments: peak systolic velocity( Vs), early diastolic velocity (Ve), late diastolic velocity (Va), time to peak systolic velocity(Ts). Then the coefficient of variation (SD/mean) of the 5 different Ts were calculated.Results The value of Vs,Ve and Va were decreased and the Ve/Va ratio was reverses three days before PTCA + stent. Compared with that before PTCA + stent,the value of Vs and Ve were increased significantly in one week ( P ＜0. 05) and one month( P ＜0.01 ) after PTCA + stent,respectively,the value of Va was not statistically significant. Ve/Va ratio was recovered in one week after PTCA treatment. Ts and Ts-SD were shorted dramatically in one week( P ＜0. 05) and one month( P ＜0.01 ) after PTCA + stent compared with that before PTCA + stent in which Ts were prolonged more than 33 ms. Conclusions QTVI can quantitatively assess the left ventricular function and the synchrony of myocardial ischemic segments, and can be used to real-time detect the changes of function and synchrony of left ventricle after PTCA and stent implantation.

Objective To investigate the relationship between the catechol-O-methyl transferase (COMT) gene polymorphism and the clinical efficacy and cognitive function of risperidone in the treatment of schizophrenia.Methods 105 cases of Han Chinese patients with schizophrenia who were treated with risperidone for 12 weeks and healthy controls of 168 cases were collected.The effect of the drug therapy with the PANSS,digit vigilance test,Raven Standard Progressive Matrices,forward and backward subtests of the digit span test were evaluated,and then the rs 165599,rs4680,rs6267,rs737865 loci in COMT gene were detected.Results (1)rs737865 was not the polymorphic locus in this sample.(2) There was statistically significant between schizophrenia patients and controls in the distribution of allele frequency and genotype frequency in rs4680 (x2=8.16,P=0.02).Haplotype GA in rs165599-rs4680 was statistically significant in schizophrenia patients and controls (x2 =4.35,P =0.04).(3) After treatment,the total score ((47.64±5.75) points),subscale scores (positive symptoms (11.66±2.90) points,negative symptoms (13.79±3.18)points,general psychotic symptoms (22.09±3.59) points) and scores of five factors model in PANSS decreased and the difference was significant (P＜0.05);the scores of digital cancellation test increased significantly compared with those before treatment(t value respectively were 12.34,10.17,4.34,all P＜0.05);the scores of forward and backward subtests of the digit span test were significantly increased compared with those before treatment (t=-5.57,P=0.00) and Raven standard reasoning test scores had increased significant (t=-19.05,P=0.00).(4) The difference of instantaneous memory score changes among rs 165599 genotypes was statistically significant after treatment (F=4.06,P=0.02).(5) The difference of negative syndromes of PANSS among rs 165599 genotypes was statistically significant after treatment (F=3.11,P=0.049).(6) The difference of negative symptoms (F=4.64,P=0.01),cognitive impairment (F=3.21,P =0.045) and instantaneous memory (F=4.86,P=0.03) among rs 6267 genotype were statistically significant after treatment.Condusion Risperidone can effectively improve the psychotic symptoms of schizophrenia patients,and promote the recovery of cognitive function.Rs165599-rs4680 haplotype GA might be risk factor for the onset of schizophrenia.

In this study, eight-month-old ectomycorrhizae of Tuber borchii with Corylus avellana were synthesized to explore the influence of T. borchii colonization on the soil properties and the microbial communities associated with C. avellana during the early symbiotic stage. The results showed that the bacterial richness and diversity in the ectomycorrhizae were significantly higher than those in the control roots, whereas the fungal diversity was not changed in response to T. borchii colonization. Tuber was the dominant taxon (82.97%) in ectomycorrhizae. Some pathogenic fungi, including Ilyonectria and Podospora, and other competitive mycorrhizal fungi, such as Hymenochaete, had significantly lower abundance in the T. borchii inoculation treatment. It was found that the ectomycorrhizae of C. avellana contained some more abundant bacterial genera (e.g., Rhizobium, Pedomicrobium, Ilumatobacter, Streptomyces, and Geobacillus) and fungal genera (e.g., Trechispora and Humicola) than the control roots. The properties of rhizosphere soils were also changed by T. borchii colonization, like available nitrogen, available phosphorus and exchangeable magnesium, which indicated a feedback effect of mycorrhizal synthesis on soil properties. Overall, this work highlighted the interactions between the symbionts and the microbes present in the host, which shed light on our understanding of the ecological functions of T. borchii and facilitate its commercial cultivation.
Colon ; Corylus ; Fungi ; Magnesium ; Mycorrhizae ; Nitrogen ; Phosphorus ; Podospora ; Rhizobium ; Rhizosphere ; Soil ; Streptomyces

OBJECTIVE To investigate the effects of triptolide （TP） exposure during pregnancy and lactation on the reproductive system development and function in male offspring of rats， providing a reference for medication safety during pregnancy and lactation. METHODS Pregnant rats were randomly divided into control group （12 rats， normal saline） and T1-T4 groups [12， 13， 14， 17 rats that received TP at 200， 400， 600， and 800 μg/（kg·d） respectively]. They were given relevant medicine/normal saline intragastrically， once a day， until the offspring were born and naturally weaned， the intragastric administration volume of each rat was consistently 2 mL. After 60 days of feeding， reproductive organ weights and coefficients were measured in male offspring， testicular and epididymal histology and sperm morphology were observed. Sperm motility， sperm count， and serum levels of gonadotropin-releasing hormone （GnRH）， follicle stimulating hormone （FSH）， luteinizing hormone （LH）， and testosterone （T） in the epididymides were analyzed. Protein expressions of glycogen synthase kinase 3α （GSK3α）， phosphorylated GSK3α （p-GSK3α）， and phosphatase 1γ2 （PP1γ2） in sperm were also determined. RESULTS Compared with the control group， the testicular and epididymal weights， serum levels of GnRH and T， the relative protein expression of PP1γ2 were significantly decreased in T1-T4 groups. Additionally， in the T2 to T4 groups， there were significant reductions in the weight and coefficient of the seminal vesicle， total number of sperm， sperm concentration， sperm motility as well as relative protein expressions of GSKα， p-GSK3α in the offspring rats. Furthermore， the epididymal coefficient in the T3 and T4 groups， the testicular coefficient， mean sperm track velocity and sperm curvature velocity in the T4 group were significantly decreased （P＜ 0.05）； the number of abnormal sperm， rate of sperm abnormality， and levels of FSH and LH in the offspring rats of the T1 to T4 groups were all significantly increased （P＜0.05）； in the offspring rats of the T1 to T4 groups， there was a decrease in the number of epithelial cells in the seminiferous tubules of the testes. Within the epididymal tissue， degenerative and necrotic changes in the epithelial cells were visible， accompanied by mild infiltration of inflammatory cells in the stroma. CONCLUSIONS TP exposure during pregnancy and lactation disrupts reproductive organ development， impairs spermatogenesis and sperm motility， as well as suppresses androgen synthesis in male offspring，thereby having a negative impact on the development of the reproductive system. These effects may be mechanistically linked to regulation of GSK3α， p-GSK3α and PP1γ2 protein expressions.

Objective To investigate the clinicopathological characteristics and the diagnosis of multilocular cystic renal cell carcinoma ( MCRCC).Methods The clinicopathological data of 19 MCRCC cases were collected and immunohistochemical staining assays were carried out .Forty-six cases of other cystic kidney lesions within the same period were collected as controls , including extensively cystic clear cell RCC (12 cases), clear cell tubulopapillary renal cell carcinoma (6 cases), tubulocystic carcinoma (2 cases), simple cortical cysts (22 cases), multilocular cystic nephroma (1 cases) and multicystic kidney (3 cases). Results The patients included 14 males and 5 females.The ages ranged from 31 to 66 years ( median age=50 years ).Most of the MCRCC cases were detected incidentally in physical examination , occasionally accompanied with hematuria , back pain or other symptoms.The follow-up period of 17 patients ranged from 6 to 170 months.All patients were alive without evidence of tumor recurrence or metastasis.Pathological findings showed that macroscopically , tumor size ranges from 1.5 to 7.0 cm in the maximum diameter , generally a entirely of various sized.The cysts contain serous , hemorrhagic or turbid fluid.Solid areas or substantially discernible mural nodules were absent; histologicallly , single layer of cuboidal and flattened epithelial tumor cells were lined in the cysts , described as clear cytoplasm , small nuclear , no nucleoli and low Fuhrman nuclear grade ( I or II).Multilayer tumor cells could be observed in a few cysts , with granular cytoplasm and small intracystic papillae formed.The clear tumor cell clusters , similar as cystic lined tumor cells, were seen within pathological fibrous in almost all cases , and significant myofibroblastic proliferation was found in 14 cases.Immunohistochemically , the cysts lined epithelial cells and the clear tumor cell clusters were positive for epithelium markers , including CKpan(19/19), EMA(16/19) and CK7 (15/19);higher percentage of CAⅨ(17/19)and PAX8(15/19) than control groups, but lower percentage of CD10 (7/19), RCC (6/19) and AMACR (2/19); and all were negative for 34βE12, CD117 and CD68.Conclusions Multilocular cysts , clear cells clusters of low Fuhrman grade within fibrous septa and capillary vessel proliferation under epithelium are important features of MCRCC.The united using of CAⅨ, CK7, CD10 and RCC is helpful for differentiating variable cystic renal tumor.MCRCC usually has an excellent prognosis, nephron sparing surgery is first recommended as a therapeutic strategy.

Objective To investigate the morphological features and immunophenotypes of eosinophilic renal tumors in order to provide references for the differential diagnosis of this tumor.Methods A cohort of 75 cases of eosinophilic renal tumors were collected.The morphological features of the tumors were observed under microscope, and the immunophenotypes of the tumors were detected using tissue microarray and immunoshistochemistry.Results There were some overlaps between the different types of eosinophilic renal tumors in morphology, but each had its distinct characteristics.Immunohistochemically, renal oncocytoma ( RO) and eosinophilic chromophobe renal cell carcinoma ( ChRCC) shared some common immumophenotypes, except for the expression of CK7, with the expression rates of 2/19 in RO and 17/20 in eosinophilic ChRCC, respectively.Eosinophilic clear cell renal cell carcinoma mainly showed positive immunostaining for Vimentin and CAⅨ, whereas negative for CK7 and CD117 in most cases ( 10/15 ).AMACR was diffusely expressed in the majority of eosinophilic papillary renal cell carcinoma ( PRCC, 10/13).Furthermore, vimentin, CK7 and CD10 were positively expressed in eosinophilic PRCC with the expression rates of 8/13, 9/13 and 6/13, respectively;while CAⅨ, CD117 and TFE3 were all negatively expressed in eosinophilic PRCC.Epithelioid angiomyolipoma generally showed positive expression of vimentin,SMA and HMB45, but negative expression of CAⅨ and CK7.Vimentin, CD10, AMACR and TFE3 were strongly expressed in XP11.2 translocation renal cell carcinoma;on the contrary, CK7, CD117and HMB45 were not expressed in the majority of the tumor.Conclusion With full understanding of the morphology of different types of eosinophilic renal tumors, the immunostaining of vimentin, CAⅨ, CK7, CD10, AMACR, CD117, TFE3 and HMB45 could play a crucial role in the differential diagnosis of these tumors.

Objective:To investigate the clinicopathological characteristics of eosionphilic Chromophobe renal cell carcinoma (eChRCC), and differences in morphology, immunophenotype and clinical prognosis betweeneChRCC, renal oncocytoma(RO) and classic Chromophobe renal cell carcinoma (cChRCC).Methods:The clinicopathologic data of 17 patients diagnosed as eChRCC from the Affiliated Hospital of Qingdao University (13 cases) and 971 Hospital of PLA Navy (4 cases) from October 2006 to February 2019 were collected. Immunohistochemical analysis was carried out to compare the immunophenotypes between 17 cases with ChRCC, 27 cases with RO and 30 cases with cChRCC.Resuls:Among the 17 patients, seven were males and ten were females, and the age ranged from 40 to 75 years (median 54 years). Clinically, 15 cases of 17 were found accidentally by physical examination. The tumor size ranged from 1.8 cm to 10.0 cm (average 5.7 cm) and the cut surface of 15 cases were solid, one case was solicl and cystic, and one was cystic. Most showed gray to red, and partially soft, gray to yellow appearances. Microscopically, most tumors presented solid growth pattern with vary number of alveolar structures (12 cases). Some were predominately characterized by cystic structure (3 cases), alveolar structure(1 case) and microcapsule structure (1 case). There were boundaries with varying degrees of clarity between tumor cells in 16 cases. The cytoplasm of tumor cells was eosinophilic and the nuclei were small round or irregular with focal perinuclear haloes in 14 cases. Large polygonal cells with light-stained cytoplasm appeared focally in 9 cases, and edematous areas with scarce tumor cells were found in 4 cases. Among 7 cases, 4 cases focally invaded peripheral renal parenchyma, 2 cases invaded adipose tissues outside the renal capsule, and 1 case presented invasion of renal sinus. Immunohistochemically, all cases were moderate to strong positive for EMA and claudin-7. CK7, CD117 and Ksp-cad were highly expressed with the expression rates of 12/17, 15/17, 14/17, respectively. Cyclin D1, AMACR, CD10, S100A1, and RCC were rarely expressed with the expression rates of 4/17, 3/17, 4/17, 1/17 and 1/17, respectively. On the contrary, all cases were negative for vimentin, CAⅨ, HMB45 and Melan A. The Ki-67 proliferation index of the 17 cases was 1%?5%. Follow-up data were available for all 17 patients from 7 to 154 months. Among them, 15 patients were alive without tumor recurrence or metastasis, one patient died of pulmonary metastasis after 31 months of surgery and one patient died of hepatic metastasis after 38 months of surgery.Conclusion:eChRCC has overlapping morphology and immunophenotype with RO. eChRCC is characterized by solid nest or alveolar structure, distinct border between tumor cells, perinuclear halos and lacking of interstitial looseness and edema. Scattered large polygonal cells with light-stained cytoplasm in tumor tissue play a significant role in the diagnosis of eChRCC. The positive expression of CK7, CD117, claudin-7 and Ksp-cad, and negative expression of cyclin D1, S100A1 are helpful to the diagnosis and differential diagnosis of eChRCC. The prognosis of eChRCC after complete surgical resection is excellent and few cases may have long-term metastasis. There is no significant difference in prognosis between eChRCC and cChRCC, but eChRCC shows better outcome than RO.