Objective To explore the biocompatibility of nano-hydroxyapatite/polyamide 66 (nHA/PA66) with human bone mesenchymal stem cells (hBMSCs) after osteogenic induction.Methods After hBMSCs were isolated and cultured in vitro,the experiment was conducted in 3 groups.Group A were hBMSCs subjected to no osteogenic induction,group B hBMSCs subjected to osteogenic induction,and group C was the composite of nHA/PA66 with hBMSCs subjected to osteogenic induction.Adhesion of the cells onto the nHA/PA66 in group C was observed by electron microscope scanning.Growth and proliferation of the cells in groups B and C were detected by MTI test at 1,2 and 3 weeks.The ability of osteogenic differentiation of hBMSCs in vitro was analyzed by alkaline phosphatase (ALP) activity and alizarin red staining.The ability of osteogenic differentiation of hBMSCs on nHA/PA66 was tested by ALP activity.Results Electron microscope scanning showed that the cells spread and attached well on the surface of the composite scaffold in group C;the proliferation of the cells in groups B and C showed no significant difference (P ＞ O.05).These suggested that the proliferation of hBMSCs was not affected by nHA/PA66.The number of mineralized nodules in group B was significantly larger than in group A (P ＜ O.05);the ALP activity of the cells in group A was significantly lower than in group B at 6 and 12 days (P ＜ 0.05);no significant differences were observed between groups B and C (P ＞ 0.05).These indicated that the hBMSCs were capable of osteogenic differentiation which was not affected by nHA/PA66.In groups B and C,the ALP activity of the cells at 12 days was significantly higher than at 6 days,indicating the ALP activity increased with increased induction time (P ＜ 0.05).Conclusion nHA/PA66 can be used as a carrier of hBMSCs in bone tissue engineering because hBMSCs can well adhere to,proliferate,and differentiate into bone on nHA/PA66 scaffolds.

BACKGROUND:Orthopedists should pay more attentions to nonunion prevention in view of nonunion treatment, that is, active interventions should be taken to avoid bone delayed union and nonunion.
OBJECTIVE:To explore the effect of composite tissue-engineered scaffold constructed by nano-hydroxyapatite/polyamide 66 (nHA/PA 66) combined with bone marrow mesenchymal stem cels to repair a femoral fracture with severe nonunion.
METHODS:Rat bone marrow mesenchymal stem cels were isolated and culturedin vitro, and then they were divided into three groups: bone marrow mesenchymal stem cels without osteogenic induction, with osteogenic induction or combined with nHA/PA 66 folowed by osteogenic induction as control group, test group or composite group, respectively. Then osteogenic differentiation of bone marrow mesenchymal stem celsin vitrowas analyzed by measuring alkaline phosphatase activity and alizarin red staining, cel adhesion on the nHA/PA 66 was observed using scanning electron microscopy, and the celgrowth and proliferation were detected by MTT assay. In the meanwhile, established Sprague-Dawley rat femur nonunion models were randomly divided into three groups: the areas of nonunion were implanted with nothing as blank control group,those were with nHA/PA 66 as simple scaffold group, and the others were with nHA/PA 66 combined with bone marrow mesenchymal stem cels as composite scaffold group. Afterwards, X-ray examination, micro-CT and Masson staining were used to evaluate the femoral healing.
RESULTS AND CONCLUSION:At 6 and 12 days after osteogenic induction, alkaline phosphatase activity in the test group was significantly higher than that in the control group; at 14 days, compared with the control group, the amount of mineralized nodules in the test group was significantly higher, which indicated that bone marrow mesenchymal stem cels after osteogenic induction could differentiate into osteoblasts. Attached cels spread wel on the scaffold with good proliferation activity, suggesting that nHA/PA 66 is suitable for cel adherence, proliferation and osteogenic differentiation. Besides, at 12 weeks after modeling, in the blank control group, no calus appeared in the nonunion region. In the simple scaffold group, the broken femur did not heal at 8 and 12 weeks after surgery. In the composite scaffold group, the broken femur did not heal at 8 weeks, but a lot of calus appeared; at 12 weeks, bone healing achieved and the scaffold was encased and absorbed.These findings demonstrate that the tissue-engineered bone scaffolds constructed by bone marrow mesenchymal stem cels and nHA/PA 66 effectively prevent bone nonunion by accelerating femoral healing in a rat femur nonunion model.

Objective To investigate the effect of Xing-Zhi-Yi-Nao (XZYN) particles on the expressions of Nogo and OMgp proteins in brain of rats after acute carbon monoxide (CO) poisoning.Methods A total of 120 Sprague-Dawley rats were randomly divided into normal group,CO poisoning group and XZYN particles treatment group (40 rats in each group).The rats in CO poisoning group and treatment group of acute CO poisoning were established by using an animal chamber,and then received hyperbaric oxygen therapy.Meanwhile,rats in treatment group were further given additional XZYN particles twice a day by gavage.At 1 day,1 week,1 month and 2 months after CO poisoning,the neurobehavioral score of rats was evaluated by a Morris water maze test and a shuttle box test,and the expressions of neurite outgrowth inhibitor (Nogo) and oligodendrocyte-myelin glycoprotein (OMgp) were investigated in rat brain tissue by immunohistochemistry staining and western blotting assay,respectively.Results Compared with those in normal control group((11.6±8.4)s,(41.8±4.4)%,(16.1±2.3)s,and (1.2±0.2)s),the escape latency in CO group was significantly prolonged ((14.1±6.1)s),and the T1/ T total was obviously decreased (23.6±2.4) %,the escape time ((26.3±3.8)s),the active escape latency ((2.3±0.3)s) were notably extended at 1 d (P<0.05).The cognitive dysfunction caused by CO poisoning was more obvious in the later stage of poisoning (from 1 week to 2 months,P<0.05).Compared with those in CO group,the escape latency was significantly shortened (from (3.5±0.6)s to (3.1±0.5)s),the T1/ T total was gradually increased (from (29.7±3.2)% to (36.7±3.2)%),the escape time (from (39.7±5.4)s to (18.1±2.0)s) and the active escape latency were obviously decreased (from (4.3±0.4)s to (2.1±0.2)s) in the later stage (>1 week) in Xing-Zhi-Yi-Nao treatment group (P<0.05).The expressions of Nogo and Omgp proteins in brain tissue in CO poisoning group were gradually increased as time went by.The increased expressions of Nogo and Omgp proteins were still observed at 1 month after CO poisoning.By contrast,XZYN particles could significantly improve cognitive function,reduce the expression of Nogo protein,and there was statistical difference compared with the poisoning group (P<0.05).However,the level of Omgp protein in XZYN treatment group was slightly lower than that in CO poisoning group,but there was no difference between the two groups (P>0.05).Conclusion The expression of Nogo and Omgp proteins may be associated with brain injury and demyelination in rats induced by CO poisoning.XZYN particles can down-regulate the expression of Nogo,and pave a way for the treatment of acute brain damage and delayed encephalopathy after CO poisoning.

Objective:To survey the status quo of job burnout among rural general practitioners and to analyze its influencing factors.Methods:A total of 2 805 rural general practitioners in three counties in southwest Shandong province were enrolled in the study. The survey was conducted with a basic information questionnaire and the Maslach Burnout Inventory-Human Service Survey (MBI-HSS). The influencing factors of job burnout were analyzed by Pearson chi-square test (χ2) and binary logistic regression model.Results:A total of 2 272 rural general practitioners completed the survey with a completion rate of 81.0%. The rates of middle or high job burnout in the dimensions of emotional exhaustion, depersonalization and reduced personal sense of achievement were 55.1%(1 251/2 272), 34.0%(772/2 272) and 91.3%(2 075/2 272), respectively. Binary logistic regression model showed that working hours/week ≥ 60 h ( OR=1.341, 95 %CI:1.057-1.702, P<0.05), monthly shift times ≥20 ( OR=1.434, 95 %CI:1.182-1.739, P<0.05), daily consultation time≥13 h ( OR=1.294, 95 %CI:1.055-1.589, P<0.05), daily consultation of 11-20 patients ( OR=1.317, 95 %CI:1.075-1.614, P<0.05), no part-time job ( OR=1.583, 95 %CI:1.201-2.087, P<0.05), insomnia ( OR=2.638, 95 %CI:2.225-3.128, P<0.05), feeling depressed at work ( OR=3.170, 95 %CI:2.661-3.776, P<0.05) were risk factors for emotional exhaustion; participation in public health services ( OR=1.485, 95 %CI:1.155-1.910, P<0.05), being married ( OR=2.273, 95 %CI:1.198-4.313), no part-time jobs ( OR=1.677, 95 %CI:1.276-2.203, P<0.05), insomnia ( OR=1.487, 95 %CI:1.211-1.827, P<0.05), and feeling depressed at work ( OR=1.642, 95 %CI:1.339-2.015, P<0.05) were risk factors for depersonalization; working hours/week ≥ 60 h ( OR=2.089, 95 %CI:1.454-3.002, P<0.05), daily consultation time ≥ 9 h ( OR=2.495, 95 %CI:1.748-3.561, P<0.05), insomnia ( OR=2.061, 95 %CI:1.519-2.796, P<0.05), and feeling depressed at work ( OR=1.894, 95 %CI:1.383-2.593, P<0.05) were risk factors for reduced personal achievement. Protective factors included monthly income ≥3 000 Yuan ( OR=0.589, 95 %CI: 0.353-0.982, P<0.05) and monthly shift of 10-19 times ( OR=0.581, 95 %CI:0.411-0.820, P<0.05). Conclusion:The overall job burnout among rural general practitioners in southwest Shandong province is serious, especially the depersonalization and the reduction of personal sense of achievement. It is suggested to improve the salary, reduce the working intensity, optimize the working hours, and pay attention to their physical and mental health for rural general practitioners to alleviate the current situation of their occupational burnout.

OBJECTIVETo compare the contents of lignin in crude and processed fruits of Schisandrae chinensis, and to explore the processing mechanism of S. chinensis fruits.

METHODContents of schisandrin, schisandrol B, deoxyschisandrin, gomisin N, gamma-schizandrin and schisandrin C were determined by high performance liquid chromatography (HPLC).

RESULTExcept the content of Schisandrol B was higher or less in processed fruits than that in the crude, the other lignin contents of S. chinensis fruits in different processed products were higher than that in the crude. The alcohol-processed product had the highest content of lignin.

CONCLUSIONThe contents of lignin have changed by different processed methods, and alcohol-processed method may be the best processed method.

Objective To assess the expression level of targeting drug-based molecular biomarkers in ovarian clear cell carcinoma(OCCC)tissues and its clinical significance.Methods A total of 63 OCCC patients included 40 primary OCCC and 23 recurrent OCCC for secondary cytoreductive surgery(SCS),who had received primary surgeries at Fudan University Shanghai Cancer Center between January, 2008 and December, 2015 were enrolled, and immunohistochemistry SP method was used to test human epidermal growth factor receptor (EGFR), human epidermal growth factor receptor-2 (HER2), aurora kinase A (AURKA), breast cancer susceptibility gene 1 (BRCA1), BRCA2 and programmed death-ligand 1 (PD-L1) protein expression in paraffin-embedded tissues. Results The positive rates of EGFR, HER2, AURKA, BRCA1,BRCA2 and PD-L1 in primary and recurrent tumor tissues were respectively 20%(8/40)vs 30%(7/23),22%(9/40)vs 35%(8/23),38%(15/40)vs 35%(8/23),42%(17/40)vs 39%(9/23),20%(8/40)vs 22%(5/23), 25%(10/40)vs 17%(4/23), and there were no significant differences between primary and recurrent OCCC (all P>0.05). χ2-test or Fisher exact analysis revealed that HER2 expression in recurrent tumor tissues had a relationship with chemoresistance (P<0.05), while the expression of other biomarkers showed no significant relationship with chemoresistance (all P>0.05). Further, Kaplan-Meier survival analysis showed that patients with HER2 and AURKA-positive expression had a significantly shorter progression-free survival time in primary OCCC(4 months vs 10 months,log-rank test,P<0.05 for HER2;and 4 months vs 10 months,P<0.05 for AURKA);and a shorter overall survival time after SCS in recurrent OCCC (10 months vs 44 months, P<0.05 for HER2;and 13 months vs 43 months, P<0.05 for AURKA). However,multivariate Cox proportional hazards regression analysis indicated that none of these 6 biomarkers was independent risk factor of progression-free survival time of primary OCCC or overall survival time after SCS for recurrent OCCC (P>0.05). Conclusion HER2 and AURKA could serve as prognostic factors in ovarian clear cell carcinoma.

OBJECTIVE: To explore the clinical and pathologic features of angioimmunoblastic T-cell lymphoma(AITL) and provide evidence for diagnosis. METHODS: Eighteen AITL patients (9 males and 9 females aged from 14 to 70 years) were retrospectively analyzed in Xiangya Hospital of Central South University from July 2002 to September 2007. RESULTS: Characteristic features at the presentation of AITL included generalized lymphadenopathy, fever, splenomegaly, and skin rashes with polyclonal hyper-gammaglobulinemia and other hematological abnormalities (such as Coombs-positive hemolytic anemia), which often involved the bone marrow and had well-described histologic features. The positive rate for CXCL13 was 93.3%. CONCLUSION Repeated lymphadenbiopsy is helpful for AITL diagnosis. Routine histological and immunohistochemical examinations (especially including CXCL13) play significant role in the diagnosis and differential diagnosis of AITL.
Adolescent ; Adult ; Aged ; Chemokine CXCL13 ; metabolism ; Female ; Humans ; Immunoblastic Lymphadenopathy ; diagnosis ; metabolism ; pathology ; Lymphoma, T-Cell, Peripheral ; diagnosis ; metabolism ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Objective To investigate the effects of N-butylphthalide (NBP) on cognitive function in acute severe carbon monoxide (CO) poisoning rats and its mechanism. Methods 120 health Sprague-Dawley (SD) rats were randomly divided into three groups (n = 40): normal control group (NC group), CO poisoning group (CO group) and NBP treatment group (NBP group). The acute severe CO poisoning model was established in a hyperbaric oxygen chamber by intoxicated with 1 000 ×10-6CO for 40 minutes, followed with 3 000 ×10-6CO for another 20 minutes, and then received hyperbaric oxygen therapy 1.5 hours once a day until sacrificed. Rats in NBP group were administered orally NBP 60 mg/kg for 2 times daily until death. NC group and CO group were treated with equal amount of pure olive oil. Four rats in each group were taken from 1, 3, 7, 14, 30 days after model setup, respectively. The cognitive function score was assessed by Morris water maze test. The changes in ultrastructure of hippocampus were observed under transmission electron microscope. The expressions of calpain 1 and Ca2+/calmodulin dependent protein kinase Ⅱ(CaMK Ⅱ) in hippocampus of brain tissue were detected by immunofluorescence staining, and the localization of the two target proteins in neurons was observed by immunofluorescence double staining. Results Compared with NC group, the escape latency at 1 day after poisoning in CO group was significantly prolonged (s: 55.6±3.2 vs. 44.5±3.5, P < 0.05), and the times of the platform crossing was significantly decreased (times: 1.3±0.8 vs. 6.6±1.2, P < 0.05);the ultrastructure of hippocampus was obviously injured; the protein expressions of calpain 1 and CaMK Ⅱ in brain tissue were significantly increased at 1 day after CO poisoning [calpain 1 (A value): 41.24±5.21 vs. 6.44±1.13, CaMK Ⅱ (A value): 56.19±5.04 vs. 9.84±1.53, both P < 0.05], and the protein expression of calpain 1 reached the peak at 3 days (A value: 59.34±6.11), the protein expression of CaMK Ⅱ reached the peak at 1 day (A value:56.19±5.04). Compared with CO group, the cognitive function was significantly improved in NBP group in the late stage of poisoning [7-30 days, escape latency (s): 40.3±1.9 vs. 49.1±3.1 at 7 days, 30.1±2.9 vs. 39.4±3.1 at 30 days;times of the platform crossing (times): 2.8±1.0 vs. 1.0±0.9 at 14 days, 3.2±0.8 vs. 1.0±0.9 at 30 days, all P < 0.05];the degree of injury of hippocampal neuron was relatively slight; the protein expression of calpain 1 in brain tissue was significantly decreased from 3 days after CO poisoning (A value: 39.63±3.03 vs. 59.34±6.11, P < 0.05), and the protein expression of CaMK Ⅱ was significantly decreased from 1 day after CO poisoning (A value: 42.22±3.84 vs. 56.19±5.04, P < 0.05). Immunofluorescence double staining suggested that calpain 1 and CaMK Ⅱ protein could not only coexist in the same cell, but also could be expressed separately in different cells. Linear regression analysis showed that the expression of calpain 1 and CaMK Ⅱ was positively correlated (R 2= 0.852, P = 0.002). Conclusions NBP treatment could maintain ultrastructure integrity of hippocampus, balance the expression levels of calpain 1 and CaMK Ⅱproteins, and significantly improve cognitive impairment induced by CO poisoning, thus play a protective role against hippocampus damage in rats with acute severe CO poisoning.

Objective: To investigate the role of JAZF1 gene rearrangement in the diagnosis and differential diagnosis of endometrial stromal sarcomas by fluorescence in situ hybridization (FISH). Methods: JAZF1 gene rearrangement was analyzed by FISH in 129 cases of ESS diagnosed from January 2008 to December 2016 including 105 cases of low-grade endometrial stromal sarcoma (LG-ESS), 21 cases of high-grade endometrial stromal sarcoma (HG-ESS) and 3 cases of undifferentiated uterine sarcoma (UUS). Sixteen cases of the related tumours in uterus were also collected as control group. The results were compared with our previous studies of JAZF1/JJAZ1 fusion gene in ESS by RT-PCR. Results: Detection of JAZF1 gene rearrangement by FISH was successfully analyzed in 144 cases. JAZF1 gene alteration was detected in 63 cases, all of which were LG-ESS, with an overall positivity of 60.6% (63/104), while no JAZF1 gene rearrangement was found in all other cases. JAZF1 gene rearrangement was present in LG-ESS with classic histology (69.3%, 52/75), smooth muscle differentiation (2/10), sex cord-like differentiation (4/5), fibromyxoid change (1/5), clear cell change (0/1), skeletal muscle differentiation (0/1), and schwannoma-like palisading pattern (0/1). The different components in all the cases of LG-ESS with variant histology had the clonal origin, with or without JAZF1 gene alteration. Compared to the results of JAZF1/JJAZ1 fusion gene by RT-PCR, the positive rate of JAZF1 gene rearrangement in LG-ESS by FISH (61.9%, 26/42) was significantly higher than that of RT-PCR (30.0%, 12/40; P<0.01). Conclusions JAZF1 gene rearrangement is present only in LG-ESS, but not in HG-ESS, UUS or other related tumours in uterus. The frequency of JAZF1 gene rearrangement varies between classic LG-ESS and different morphologic variants. It is frequently, but not consistently, present in classic LG-ESS and less often positive in variant cases. The results of JAZF1 gene alterations in LG-ESS with different morphologic variants support the contention that the endometrial stromal and their variant morphologic components have the same clonal origin. Detection of JAZF1 gene rearrangement by FISH is very useful for the diagnosis and differential diagnosis of ESS.

Objective: To investigate the expression of SMARCA4 (BRG1) and SMARCB1 (INI-1) protein in endometrial dedifferentiated carcinoma (DDC) and undifferentiated carcinoma (UDC), and their correlation with clinicopathologic features. Methods: Clinicopathological information was gathered for 26 cases of DDC and UDC and consulting hospitals from January, 2006 to December, 2018 in Fudan University Shanghai Cancer Center, including 10 cases of DDC and 16 cases of UDC. Morphologic features and diagnosis were reviewed by two pathologists. Immunohistochemistry for expression of BRG1 and INI1 protein was performed. The correlations with clinicopathologic features were analyzed. Results: BRG1 and INI1 loss were present in 14 of 26 cases of DDC/UDC, including 12 BRG1-deficient cases and 2 INI1-deficient cases, respectively. Six cases demonstrated variable amounts of rhabdoid cells in 14 BRG1/INI1-deficient cases, and only 1 case showed rhabdoid cells in the 12 intact expression cases. However, there was no significantly statistical difference (P=0.060). Age, invasive depth, lymph node status and FIGO stage were not associated with the expression of the BRG1 and INI1 (P=0.437, P=0.672, P=0.242, P=0.348). Remarkably, the BGR1/INI1-deficient patients had worse survival than those with intact expression (4.7 vs. 22.9, P=0.033). Conclusion BRG1/INI1-deficient is observed in approximately half of DDC and UDC. Identification of these tumors is clinically relevant due to their more aggressive behavior and poor prognosis. Hence, BRG1 and INI1 immunohistochemical stains should be performed for DDC and UDC in order to help the pathologists to distinguish these tumors from other carcinomas, and to predict the clinical prognosis.