Objective:To investigate the genomic manifestation and pathogenesis of osteosarcoma with different relapse pattens, which were respectively initially presented with bone metastasis or pulmonary metastasis.Methods:From May 1, 2021 to October 1, 2021, 38 fresh tumor specimens and some paraffin-embedded specimens of high-grade osteosarcoma were collected in Peking University People's Hospital, including 29 males and 9 females, aged 19.6±2.2 years (range, 6-61 years). Among the 38 cases, 12 cases had initial bone metastasis (group A) and 26 cases had initial lung metastasis (group B), of which 15 cases (40%, 15/38) had paired specimens of primary and metastatic lesions. Based on Illumina NovaSeq 6000, we analyzed whole-exome sequencing (WES) as well as transcriptome for osteosarcoma with paired samples in different relapse patterns. During all their treatment courses, we also collected their paired samples to reveal these tumors' evolution. We sought to redefine disease subclassifications for osteosarcoma based on genetic alterations and correlate these genetic profiles with clinical treatment courses to elucidate potential evolving cladograms.Results:We found that osteosarcoma in group A mainly carried single-nucleotide variations (83%, 10/12), displaying higher tumor mutation burden [4.9 (2.8, 12.0) & 2.4 (1.4, 4.5), P=0.010] and neoantigen load [743.0 (316.5, 1,034.5) & 128.5 (49.0, 200.5), P=0.003], while those in group B mainly exhibit structural variants (58%, 15/26). The mutation spectrum showed that there was a significant difference in age-related gene imprinting 1 between the bone metastasis group and the lung metastasis group ( P=0.005). Samples were randomly selected from group A (3 patients) to investigate immunologic landscape by multiplex immunohistochemistry, from which we noticed tertiary lymphatic structure from one patient from group A. High conservation of reported genetic sequencing over time was found in their evolving cladograms. Conclusion:Osteosarcoma with mainly single-nucleotide variations other than structural variants might exhibit biological behavior predisposing toward bone metastases with older in age as well as better immunogenicity in tumor microenvironment.

Objective To investigate the effects of PDGF-D/PDGFR-β signaling on interstitial collagen hyperplasia and fibroplasia of myocardium following cardiopulmonary resuscitation (CPR)in rats.Methods A total of 90 male Sprague-Dawley (SD) rats were randomly (random number) assigned into 5 groups:sham,control,Imatinib,Ad-GFP and Ad-PDGFR groups.Each group was further subdivided into post-resuscitation (PR) 4 h and PR 72 h groups.The ventricular fibrillation cardiopulmonary resuscitation (CPR) model was used.Cardiac collagen volume fraction (CVF) was measured with Sirius red staining.Platelet derived growth factor D (PDGF-D) was detected by enzyme-linked immunosorbent assay (ELISA).Collagen Ⅰ,α smooth muscle actin (α-SMA),vimentin and PDGF receptor β (PDGFRβ) were detected by Western blot.The data were analyzed by a two-way ANOVA and Bonferroni test.P ＜0.05 was considered statistically significant.Results (1) At PR 72 h,the CVF value in the control group was significantly higher than that in the sham group [(11.24 ± 0.31) vs (1.65 ± 0.19),t =10.81,P ＜0.05] and in the Ad-PDGFR group significantly higher than that in Ad-GFP group [(25.60 ±4.09) vs (10.73 ± 2.42),t =16.77,P ＜0.05],respectively.Compared to the control group,the level of CVF in the Imatinib group significantly decreased [(5.11 ± 0.29)vs (11.24 ± 0.31),t =3.892,P ＜0.05].(2) At PR 4 h,compared with the sham group,the expression of serum PDGF-D was greatly increased in the control,Imatinib,Ad-GFP and Ad-PDGFR groups (all P＜0.05).At PR 72 h,serum PDGF-D was continuously significantly higher in the Ad-PDGFR group when compared with the sham group [(296.46± 30.82) pg/mL vs (93.74 ± 5.43) pg/mL,t =7.755,P ＜0.05].(3) At PR 72 h,the expression of Collagen I,α-SMA,Vimentin and PDGFR-β in the control and Ad-PDGFR groups were significantly increased than those in the sham and Ad-GFP groups,respectively (all P＜0.05).However,compared with the control group,Imatinib significantly decreased the expressions of the above proteins (all P＜0.05).Conclusions The interstitial collagen hyperplasia and fibroplasia of myocardium at 72 h following CPR in rats might be related to the increased levels of PDGF-D in serum and PDGFR-β in myocardium.

OBJECTIVETo determine the frequency of spinocerebellar ataxia type 31 (SCA31) related mutations among patients from mainland China.

METHODSFor a cohort of molecularly unassigned patients comprised of 295 SCA patients (including 98 probands from families featuring autosomal dominant SCA and 197 sporadic cases) and 81 patients with hereditary spastic paraplegia (HSP) (including 23 probands from families with autosomal dominant HSP and 58 sporadic cases),TGGAA pentanucleotide expansion insertional mutation of the BEAN/TK2 gene was detected using repeat-primed PCR followed by capillary gel electrophoresis.

RESULTSNo TGGAA pentanucleotide insertion expansion in BEAN/TK2 gene was identified in the above cohort.

CONCLUSIONSCA31 is an extremely rare subtype of SCA and should not be included in routine genetic screening in mainland China.

Objective To investigate the effects of real-time feedback devices on chest compression quality test in non-medical staff during cardiopulmonary resuscitation (CPR) training.Methods A total of 120 volunteers were recruited and trained according to American Heart Association Guidelines Update for Cardiopulmonary Resuscitation and Emergency Cardiovascular Care set in 2015.CPR performance with compression for six minutes was tested on a manikin.Volunteers were randomized into 3 groups.Group A was tested without any feedback.Group B was self-corrected in compression quality(include compression depth、rate and rebound of chest wall) using a real-time feedback device (Link CPR).Group C was guided with a metronome.All compression data were collected via WiFi signal and stored.Results Significantly better mean chest compression depth was achieved in group B than that in group A and C(5.38 ± 0.483 cm vs.4.42 ± 0.572cm and 4.25 ± 0.843 cm,P ＜ 0.05).Significantly better compression rate were observed in both group B and C than that in group A (113.4 ± 5.9 and 109.0 ± 6.8 compressions/min vs.129.6 ± 8.3 compressions/min,P ＜ 0.05).Significantly less rebounding were observed in both group B and C compared with group A (56.10 ± 32.3 and 68.30 ± 28.8 compressions vs.174.30 ± 38.8compressions,P ＜ 0.05).Pearson correlation analysis confirmed the compression rate was positively correlated with the numbers of rebounding (r=0.776,P＜0.01).Significant statistical difference in accuracy was observed among the groups (9.8％ vs.72.9％ vs.58.5％,P ＜ 0.05).Conclusions In CPR training test real-time feedback device contributes to the improvement of chest compression quality through self-adjustment of compression depth,rate and rebound.

Objective: To investigate the relationship between menopausal status at different FPG levels and the risk of new onset of impaired fasting glucose (IFG) and type 2 diabetes mellitus (T2DM). Methods: Female subjects were selected from the females who joined in the epidemiological survey between May and September in 2009 in six counties of Guanlin Village and Xushe Village in Yixing City in Jiangsu Province by cluster sampling method. Subjects with diabetes at baseline or pre-menopause with age above 65 years old or surgical menopause were excluded. Finally there were 2 084 eligible subjects included in this study. According to FPG at baseline, subjects were categorized into two groups as normal FPG group (FPG<5.6 mmol/L) and IFG group (5.6≤FPG≤6.9 mmol/L). Follow-up study was conducted from May in 2014 to January in 2016. Cox regression model was used to investigate the association between menopausal factors and IFG and T2DM among groups based on FPG. Results: The average age of subjects at baseline was 58.08 (51.74-65.82) years old, and the number of post-menopausal women was 1 631 (78.3%). The number of subjects in normal FPG group was 1 569 (75.3%), and in IFG group was 515 (24.7%). There were 104 subjects with new onset T2DM among which 34 subjects from normal FPG group and 70 subjects from IFG group. And there were 199 subjects with new onset IFG in normal FPG group. Among subjects with normal FPG, the incidence density of IFG in post-menopausal women (2 752/100 000 person-years) was statistically higher than that in premenopausal women (1 670/100 000 person-years) (P<0.001). After age and BMI adjusted, post-menopausal women had a higher risk of having IFG in normal FPG subjects with hazard ratio (HR) at 2.60 (P<0.001). Among subjects with normal FPG, the risk of new onset IFG decreased in post-menopausal women with menopause age increasing after age and BMI adjusted, with HR at 0.96 (P=0.046). No statistical association was found between menopausal factors and risk of T2DM either in the overall subjects or in the subgroups(P>0.05). Conclusion Menopause can increase the risk of IFG incidence in subjects with normal FPG. The incidence of IFG decreases with the menopause age increasing.

Objective To learn current status and influentcing factors of post competency among preventive medicine graduates from a medical university,in order to provide evidence for the improvement of post competency preventive medicine graduates.Methods We adopted stratified sampling strategy,chose 1 provincial,1 municipal and 2 county level CDCs,used self-developed questionnaires to survey preventive medicine graduates from a specific medical university.Clustering analysis,correlation analysis,Logistic regression and rank-sum test were applied to analyze data.Results Among all respondents,26 (40.63％) reported that they were totally competent on their posts,the rest 38 (59.37％) reported ordinary post competency.We found 7 factors that were significantly associated with post competency:fondness of the job,whether proactive on the job,whether responsible to the job,et al.Conclusion Post competency of preventive medicine graduates from this particular medical university exceeded preventive medicine graduates from Xinjiang Medical University.However,there still exists some problems for improvement.We recommend that universities and working units should improve post competency of preventive medicine students based on those influential factors we identified.

Objective To evaluate the efficacy and quality of life of segmental bowel resection for bowel endometriosis. Methods Totally 62 symptomatic patients with bowel endometriosis undergoing segmental bowel resection were recruited. A visual analogue scale (VAS) and the 36-item short form health survey (SF-36) questionnaire were administered before and at least 1 year after surgery, respectively. Pregnancy rates were also recorded. Results Sixty-two patients in total underwent follow-up ranging from 12 to 74 months. All patients complained of obvious pain symptoms, including dysmenorrhea, dyspareunia, pain on defecation and chronic pelvic pain. The relief of dysmenorrhea (2.9 ± 2.2 versus 7.5 ± 2.9), dyspareunia (0.7 ± 0.5 versus 4.3 ± 2.2) and pain on defecation (1.6 ± 0.7 versus 7.3 ± 1.9) after surgery was statistically significant (all P<0.01). The scores for all 8 domains of the SF-36 questionnaire were significant improved after segmental bowel resection (all P<0.01). The complication rate was 45% (28/62), including 18 cases of urinary retention, 4 rectovaginal fistulas, 2 cases of vaginal dehiscence, and 1 case each of thrombogenesis, pelvic abscess and general peritonitis. All of the patients with complications recovered well throughout follow-up. The postoperative pregnancy rate of the previous infertile patients was 6/10. Among the 6 gestational cases, 2 had labour, 2 underwent caesarean sections, one had a spontaneous natural abortion, and one underwent uterine curettage. Conclusion Segmental bowel resection could significantly relieve pain and improve quality of life for patients with bowel endometriosis.

This study is designed to explore the possible effects of Hemerocallis citrina baroni flavonids (HCBF) on liver fibrosis induced by CCl4 in rats. The liver fibrosis model was induced by CCl4, and HCBF were administered by gastric perfusion at 25 and 50 mg x kg(-1) qd for 50 days, while the contents of alanine transaminase (ALT), aspartate aminotransferase (AST), gamma glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), superoxide dismutase (SOD), maleic dialdehyde (MDA) and transforming growth factor-β1 (TGF-β1) were measured and the contents of PINP were measured in liver tissue, and the expression of TGF-β1 were observed by immunohistochemisty and Western blot. The pathological changes of liver tissue were examined by HE. The results showed that HCBF (25, 50 mg x kg(-1)) improved the liver function significantly through reducing the level of ALT, AST, GGT and ALP (P < 0.05 or P < 0.01), and increasing the content of SOD (P < 0.01), while reducing the content of MDA (P < 0.05 or P < 0.01), the expression of TGF-β1 (P < 0.05) and the content of PINP (P < 0.05). The results suggest that HCBF (25, 50 mg x kg(-1)) may inhibit the liver injury induced by CCl4 by decreasing the oxidative stress.

OBJECTIVETo explore the effects and action mechanism of electroacupuncture (EA) on Parkinson's disease (PD).

METHODSForty-eight healthy male SD rats were randomly divided into a normal group, a sham operation group, a model group and an EA group, 12 rats in each one. Rats in the model group and EA group were treated with subcutaneous injection of rotenone (1mg/kg, dissolved in dimethyl sulfoxide and 0. 9 % normal saline) on neck and back for 40 days to establish rat model. Rats in the sham operation group were treated with injection of identical dose of dimethyl sulfoxide and 0. 9 %o normal saline at identical location which did not contain rotenone. After model establishment, rats in the EA group were treated with EA at "Fengfu" (GV 16) and "Taichong" (LR 3) with continuous wave (2 Hz, 1 mA), which was given 20 min per time, once a day for consecutive 28 days. Rats in the remaining groups were treated with fixation and immobilization without any other intervention. The rats behavioristics changes were observed and scored; immunohisto-chemistry was adopted to test the expression of tyrosine hydroxylase (TH); fluorescence spectrometry was used to detect the activities of 20 S β1, β2, β5; western blot method was applied to measure the expression of 20S proteasome and its a subunit.

RESULTSCompared with the normal group and sham operation group, there was significant change of behavioristics in the model group, and TH positive neuron counting was obviously reduced; after treatment, the behavioristics score in the EA group was lower than that in the model group (P<0. 05), and TH positive neuron counting was significantly increased (P<0. 05). Compared with the normal group and sham operation group, the activities of 20 S β1, β2, β5 in model group were significantly reduced (all P<0. 01), and those in the EA group were higher than those in the model group (P<0. 01). Compared with the normal group and sham operation group, the expression of 20S proteasome and its a subunit was reduced in the model group, and that in the EA group was higher than that in the model group (P<0. 05).

CONCLUSIONEA could improve the loss of dopaminergic neurons induced by rotenone to prevent and treat PD, which is likely to be related with protecting the activity and expression of proteasomes in substantia nigra.

Animals ; Disease Models, Animal ; Electroacupuncture ; Humans ; Male ; Parkinson Disease ; enzymology ; therapy ; Proteasome Endopeptidase Complex ; metabolism ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; enzymology

AIM:To explore the effect of Vaccinium vitis procyanidin on the growth of glioma cells .METH-ODS:Glioma C6 cells were cultured and divided into control and 10, 20 and 40μg/L Vaccinium vitis procyanidin groups . The influence of Vaccinium vitis procyanidin on the growth of C 6 cells was measured by MTT assay and the observation un-der inverted microscope .The apoptotic rate was detected by Annexin V/PI staining .The protein expression of Bcl-2 and Bax was determined by immunocytochemistry .The protein levels of Bcl-2, Bax and caspase-3 were also examined by West-ern blotting .RESULTS:The growth of C6 glioma cells was inhibited by Vaccinium vitis procyanidin at concentrations of 10, 20 and 40 μg/L.The growth was significantly inhibited in 40 μg/L Vaccinium vitis procyanidin group at 24 h and 48 h, and in 20 and 40 μg/L Vaccinium vitis procyanidin groups at 72 h (P<0.01).The density of the cells was decreased when the concentration of Vaccinium vitis procyanidin increased .The apoptotic rate was increased when the concentration of Vaccinium vitis procyanidin increased either .The expression of Bcl-2 was decreased and Bax was increased after 10, 20 and 40 μg/L Vaccinium vitis procyanidin treatments .The ratio of Bax/Bcl-2 was increased when the dose of Vaccinium vitis pro-cyanidin increased (P<0.05 or P<0.01).The expression of Bcl-2 was decreased (P<0.01), and Bax and caspase-3 were increased after 10, 20 and 40 μg/L Vaccinium vitis procyanidin treatments .The ratio of Bax/Bcl-2 was increased when the dose of Vaccinium vitis procyanidin increased (P<0.01).CONCLUSION:Vaccinium vitis procyanidin inhibits the growth of glioma cells by down-regulating Bcl-2 protein and up-regulating Bax protein to activate caspase-3, thus indu-cing apoptosis .