Objective To explore whether CD44 +/CD24 + expressing cervical cancer cells are resistant to X-ray irradiation and investigate the underlying mechanism.Methods Cervical cancer cell line (Siha) was cultured in vitro and the CD44 +/CD24 + expressing cells were sorted with a flow cytometer.The cells were irradiated with 8,16 and 30 Gy of 6 MV X-rays.Colony formation test was used to evaluate the radiosensitivity of CD44 +/CD24 + expressing cervical cancer cells.Cell morphology was observed by electronmicroscopy,cell apoptosis was analyzed with a flow cytometer and also verified with a DNA ladder assay.Gene expression was determined by RT-PCR.Results After radiation,the ratio of CD44 +/CD24 + cells significantly increased.Compared to Siha cells,the radiosensitivity of CD44 +/ CD24 + cells decreased (t =93.99-400.45,P ＜0.05),and the expressions of bcl-2,survivin and Oct4 mRNA increased in CD44 +/CD24 + cells (t =221.35,941.65,82.27,P ＜0.01).Both apoptotic body and specific DNA ladder pattern were observed in cells but not in the CD44 +/CD24 + Sihacells which had no obvious morphological changes of apoptosis.Conclusions The CD44 +/CD24 + expressing cervical cancer cells are resistant to X-rays due to expression of anti-apoptosis factors.

Objective:To observe and analyze the surface electromyography activity of quadriceps femoris of knee osteoar-thritis(KOA)patients. Method:The surface electromyography of vastus lateralis,rectus femoris,and vastus medialis were recorded and analyzed in 30 KOA patients and 30 normal people during isokinetic knee flexion or extension at veloci-ties of 60 °/s,90 °/s or 180 °/s.We also record clinical indicators(muscle thickness and degree of knee pain)of the KOA group and evaluate the correlation between electromyographic data and clinical indicators. Result:Compared with the healthy control group,the KOA group had significantly lower knee extensor mo-ment during isokinetic knee flexion or extension at velocities of 60°/s(P＜0.001),90°/s(P＜0.01),or 180°/s(P＜0.01).Statistics showed that there were significant differences in the median frequency difference rate of vastus lateralis(P＜0.05)and root mean square value of rectus femoris(P＜0.01)between the healthy control group and the KOA group under 60°/s angular velocities.There was a significant difference in the root mean square value of rectus femoris(P＜0.05)between the two groups at velocities of 90°/s.Statistics revealed that the median frequency difference rate(P＜0.05)and root mean square value(P＜0.05)of rectus femoris had changed significantly at the velocities of 180°/s.The results of both multiple regression and random forest algo-rithm showed that knee extensor moment was the strongest predictor between electromyographic characteristics and clinical indicators. Conclusion:The strength of quadriceps femoris in KOA patients was decreased.Thus,the patient may need more muscles to complete the same action,which was easy to cause muscle fatigue.The knee extensor moment was closely related to the quadriceps femoris thickness and the degree of knee pain.It offers important advantag-es for the diagnosis and assessing the severity,which may provide the way for the future study of KOA.

Objective To analyze the result and adverse reactions of radiation therapy in patients with pelvic recurrence following cervical cancer postoperative.Methods A retrospective analysis of 147 patients with pelvic recurrence after surgical treatment of cervical cancer in the Fourth Hospital of Hebei Medical University from August 2004 to December 2016 was performed.All patients received radiotherapy with or without chemotherapy.According to different clinical factors and pathological factors,Logistic regression analysis was used to analyze the factors influencing radiotherapy outcomes in patients with pelvic recurrence after cervical cancer surgery.The Kaplan-Meier method was used to analyze the survival rate,and the corresponding survival curve was drawn.The survival rate and prognosis related factors were compared by using the log-rank test.The COX proportional hazards regression model was used for multivariate analysis of statistically relevant factors in univariate analysis.After treatment,toxicities were analyzed using chi-square test.Results The median follow-up time was 33.2 months.95％ of the patients completed radiation therapy with a dose of ≥ 67 Gy (median radiotherapy dose),and 91 patients (61.9％) had complete remission (CR).The 5-year local control (LC),progression-free survival (PFS),distant metastasis-free survival (DMFS),and overall survival (OS) were 63.6％,56.0％,73.9％,and 55.0％,respectively.Univariate logistic regression analysis showed that FIGO staging (stage 0-ⅠB and ⅡA-ⅡB),pelvic sidewall involvement,and recurrent tumor volume were associated with complete remission (P＜0.05).Multivariate statistical analysis found that FIGO staging and pelvic sidewall invasion were independent factors influencing the efficacy and survival of patients with pelvic recurrence after cervical cancer surgery (P＜0.05).Patients with pelvic wall invasion after cervical cancer surgery had a higher incidence of ≥ grade 2 proctitis than those without pelvic walls involved,which were 26.9％ and 16.7％,respectively.Conclusions This study shows that after the surgical treatment of cervical cancer patients with pelvic recurrence can be tolerated by toxicities after radiation therapy.In addition,the incidence of toxicities in patients with pelvic wall invasion was significantly higher than those without pelvic wall invasion.Preoperative staging and the pelvic wall involvement are independent influencing factors influencing the effect of radiotherapy and long-term prognosis in patients with pelvic recurrence after cervical cancer surgery.

Objective To investigate the characteristics of radiation resistance of cervical cancer cells,and to explore the mechanism of tumor recurrence and migration.Methods Cervical cancer cells (Siha) were fractionally irradiated to get radioresistant subpopulation.CD44 +/CD24 + Siha cells were sorted with a flow cytometry.Colony-formation tests and tumor xenografts tests were used to evaluate the " stemness" of resistant cells.Stem cell markers were studied using fluorescence-activated cell sorting analyses.Migration and invasiveness were assessed by a Transwell test.Gene and protein expressions were determined by RT-PCR and immunoblotting assay,respectively.Results Radiation-resistant Siha cells and CD44 +/CD24 + Siha cells expressed more antiapoptotic protein Bcl-2(t =205.26,198.17,P ＜0.05),apoptosis-inhibitory protein Survivin (t =896.62,765.34,P ＜ 0.05) and stem cell markers of OCT-4 and ABCG2 (t =92.13,81.26,220.45,216.32,P ＜0.05).They were more tumorigenic in vitro and in vivo,showed phenotypic and molecular changes of EMT,and had higher abilities of invasion and migration.Conclusions The radioresistant cervical cancer cells and CD44 +/CD24 + cervical cancer cells are similar to CSCs and undergo EMT,suggesting that radiation resistance-induced EMT is linked to the generation of CSCs.

【Objective】 To research the genetic polymorphism of HPA 1-6/10/15/21 in platelet donors in Zhongshan area, and establish a gene bank of platelet donors with HPA locus. 【Methods】 The HPA 1-6/10/15/21 system genotyping was performed by Real time fluorescent PCR combined with TaqMan probe technology on 192 platelet donors in Zhongshan area, and the genotype frequency and gene frequency were calculated. 【Results】 Only HPA-aa genotype was found within HPA-4/10, and no allele HPA-b had been detected. The majority of HPA-1, 2, 5, 6 and 21 genotypes were aa. HPA-3 and HPA-15 showed high heterozygosity, with genotype frequency of 0.307 3, 0.494 8 and 0.197 9 for HPA- 3aa, HPA-3ab and HPA-3bb, while 0.270 8, 0.505 2 and, 0.224 0 for HPA -15aa, HPA-15ab and HPA-15bb, respectively. 【Conclusion】 The distribution characteristics of HPA 1-6 /10/15/21 of platelet donors in Zhongshan shows regional differences compared with similar researches from other regions. The establishment of HPA gene bank is helpful to avoid alloimmunization caused by incompatible platelet transfusion.

【Objective】 To investigate the frequency of CD36 deletion and gene mutation in voluntary blood donors of Zhongshan city, and to explore the possibility of establishing local CD36 negative platelet donor bank. 【Methods】 Platelet CD36 antigen was detected by ELISA in 1 654 voluntary blood donors.Some of the negative samples were confirmed by flow cytometry, and genotyping was also performed. 【Results】 Platelet CD36 antigen was negative in 27 cases, accounting for 1.6% (27/1654), among which 1.6% (18/1149) were males and 1.8% (9/505) were females.No significant difference was noticed between males and females in CD36 antigen deletion cases (P>0.05). Fifteen CD36 negative samples were randomly selected, genotyped and sequenced, with type I deletion in 1 case[ 6.7% (1/15)], type Ⅱ deletion in 14 cases[ 93.3% (14/15)], and gene mutation in exon 3-14 detected in 8 cases. 【Conclusion】 The frequency of platelet CD36 antigen deletion in Zhongshan is comparable to that in other southern regions of China.The establishment of CD36 negative platelet donor bank is conductive to improve the effectiveness of platelet transfusion.

BACKGROUNDBone marrow hematopoietic function suppression is one of the most common side effects of chemotherapy. After chemotherapy, the bone marrow structure gets destroyed and the cells died, which might cause the hematopoietic function suppression. Heme oxygenase-1 (HO-1) is a key enzyme of antioxidative metabolism that associates with cell proliferation and resistance to apoptosis. The aim of this study was to restore or resist the bone marrow from the damage of chemotherapy by the HO-1 expression of mouse mesenchymal stem cells (mMSCs) homing to the mice which had the chemotherapy-induced bone marrow suppression.

METHODSOne hundred and sixty female Balb/c mice (6-8-weeks old) were randomly divided into four groups. Each group was performed in 40 mice. The control group was intraperitoneally injected for 5 days and tail intravenously injected on the 6th day with normal saline. The chemotherapy-induced bone marrow suppression was established by intraperitoneally injecting cyclophosphamide (CTX) into the mice which performed as the chemotherapy group. The mMSCs were tail intravenously injected into 40 chemotherapically damaged mice which served as the mMSCs group. The difference between the HO-1 group and the mMSCs group was the injected cells. The HO-1 group was tail intravenously injected into the mMSCs that highly expressed HO-1 which was stimulated by hemin. The expression of HO-1 was analyzed by Western blotting and RT-PCR. Cell proliferation was measured using the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Histopathologic examinations were performed 1 week after injection.

RESULTSCompared with the control group, the expression levels of HO-1 mRNA and protein were significantly higher in the HO-1 group (all P < 0.05), even obviously than the mMSCs group. CTX treatment induced apoptosis and inhibited proliferation. After injected, the white blood cell (WBC), red blood cell (RBC) and platelet (PLT) declined fast and down to the bottom at the 7th day. The bone marrow structure was destroyed incomplete. In vitro, the survival rate of cells in chemotherapy group was less than 50% after 24 hours. In contrast, mMSCs could do a favor to the cellular cleavage and proliferation. They slowed down the cell mortality and more than 50% cells survived after 24 hours. The effects of blocking apoptosis and bone marrow recovery could be more effective in the HO-1 group. In the HO-1 group, it had observed that the bone marrow structure became complete and the hemogram closed to normal at 7th day.

CONCLUSIONSHO-1 played an important role in promoting the recovery of CTX-induced hematopoietic damage. We suggest that HO-1 is able to restore the functions of chemotherapy-induced hematopoietic damage.

Animals ; Apoptosis ; drug effects ; Blood Platelets ; drug effects ; Blotting, Western ; Bone Marrow ; drug effects ; enzymology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclophosphamide ; toxicity ; Erythrocytes ; drug effects ; Female ; Heme Oxygenase-1 ; genetics ; metabolism ; Leukocytes ; drug effects ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; enzymology ; physiology ; Mice ; Mice, Inbred BALB C ; Reverse Transcriptase Polymerase Chain Reaction

【Objective】 To investigate the frequency of platelet antibodies in voluntary blood donors and patients in Zhongshan, Guangdong Province, and to study the specificity and cross-matching of platelet antibodies. 【Methods】 Platelet antibodies of blood donors and patients were screened by solid-phase immunoadsorption (SPIA), rechecked by flow cytometry (FCM), and antibody specificity was identified by PakPlus enzyme immunoassay, and platelet cross-matching was simulated by SPIA. 【Results】 A total of 1 049 blood donor samples and 598 patient samples were tested, with 6 (0.57%) and 49 (8.19%) samples positive for SPIA,respectively(P<0.05); In SPIA positive samples, the positive concordance rate of FCM in blood donors and patients was 100% vs 95%, and that of enzyme immunoassay was 100% vs 88%. Among the initial screening positive samples of blood donors, 5 were anti-HLA Ⅰ antibodies, accounting for 83%, and 1 was anti CD36 antibody, accounting for 17%, with an incidence rate of 0.10%. Among the 14 samples of enzyme immunoassay positive patients, 2 were anti-GP Ⅱb/Ⅲa, 1 was anti-GP Ⅱa/Ⅱa, 8 were anti HLA Ⅰ, and 3 were mixed antibodies (HLA Ⅰ, GP Ⅱb/Ⅲa, GP Ⅰa/Ⅱa). According to the types of antibodies, HLA Ⅰ antibodies were the most common, accounting for 65% (11/17), followed by HPA related anti GP, accounting for 35% (6/17). The majority of patients had a platelet antibody positive typing rate below 30%, accounting for 71.4% (10/14). 【Conclusions】 The positive rate of platelet antibody of patients in Zhongshan area is significantly higher than that of voluntary blood donors, and most of them are anti-HLA Ⅰ and anti-GP, and the incidence of anti-CD36 is extremely low. Therefore, it is necessary to establish a known platelet antigen donor bank, and at the same time, carry out platelet antibody testing and matching of patients, which is helpful to solve the issue of platelet transfusion refractoriness.