Objective To detect the expression of stem-cell related factors Nanog and CD44 in spheroid body-form?ing cells of gastric cancer cell line MKN-45. Methods The gastric cancer cell line MKN-45 was used to culture spheroid bodies in non-adherent condition in a serum-free medium supplemented with epidermal growth factor (EGF) and basic fibro?blast growth factor (bFGF). Using Western blot analysis, immunofluorescence staining and quantitative real time polymerase chain reaction(qRT-PCR), the expression levels of stem cell-related genes Nanog and CD44 were studied. Results In this study, we observed that MKN45 cells formed spheroid bodies in non-adherent condition in a serum-free medium, and the levels of Nanog and CD44 mRNA expression in spheroid body-forming cells were 2.34 ± 0.22 and 1.18 ± 0.04,respectively, which were higher than those in parental cells (1.00±0.00 and 1.00±0.05). The levels of Nanog and CD44 protein expression in spheroid body-forming cells were 0.18±0.02 and 0.24±0.04, respectively, which were significantly higher than those in pa?rental cells (0.07±0.02 and 0.18±0.01, P<0.05). Nanog protein was positively stained within the perinuclear and cytoplasm of the spheroid body-forming cells, and CD44 was positively stained mainly in the membrane. Dual staining of Nanog/CD44 indicated that the embryonal protein Nanog was co-localized with CD44 in the spheroid body-forming cells. Conclusion Spheroid body-forming cells developed from human gastric cancer cell line MKN-45 in serum-free medium supplemented with EGF and bFGF show characteristics of cancer stem cell (CSC). The cells co-expressed of CD44 and Nanog maybe a phe?notype of gastric CSCs.

status.Conclusion HPV6 and HPV-16 were the most two popular HPV types in the whole population,while HPV-16 was the most common type in CIN2+ population.HPV-16 seroprevalence increased with severity of cervical intraepithelial neoplasia.

Objective To observe the clinical effect of repairing the defect after extended resec-tion of lower lip cancer by modified lip and cheek fan-shaped flap .Methods Six cases of lower lip cancer were treated .The defect of the lower lip was about 1/3-1/2 after 1 cm of enlarged resection outside the margin of the mass .The defect was repaired with modified lip and cheek fan-shaped flaps . The morphology and function were observed 3-36 months after operation .Results All patients healed in one stage ,and the shape and function of lower lip recovered well without complications .Conclusions Lower lip defect about 1/3-1/2 of patients with modified lip and cheek fan-shaped flap is an ideal method of restoration and reconstruction ,with satisfactory results after surgery .

OBJECTIVESTo investigate the risk factors for cervical cancer in the areas of high incidence, and provide evidence for current intervention of cervical cancer.

METHODSIn the areas of Xiangyuan County, Shanxi Provicne with high incidence of cervical cancer, 1 997 women were interviewed using a questionnaire, including baseline information, menstrual, marital and pregnancy histories, sexual behavior, health habits, contraception, medical history and family history of cancer, etc., after its screening with six kinds of methods. All subjects, including 84 cases with pathological diagnosis of greater than cINI, and 1 784 cases with pathological diagnosis of normal, were tested for high-risk HPV.

RESULTSThe overall rates of HPV infection were 20.8% (415/1 997) in high-risk subjects, 97.7% and 14.2% in the cases and control groups, respectively. Univariate analysis showed that risk factors with statistical significance included high-risk HPV infection, age at first sexual intercourse, history of pregnancy and abortion, the number of sexual partners and family history of cancer. Analysis with non-conditional logistic regression model revealed high-risk HPV infection, multiple sexual partners and family history of cancer associated obviously with occurrence of cervical cancer. In addition, there was significantly positive relationship between HPV infection, which increased with the number of sexual partners, and extramarital sexual activity both in males or females.

CONCLUSIONSThe main risk factor for cervical cancer in this region was high-risk HPV infection, which related to sexual behavior, hygienic habits during menstruation and puerperium. It was particularly important to detect and treat precancerous lesions and to implement behavior modification. In addition, further research on genetic susceptibility was suggested.

Abortion, Induced ; Adult ; Analysis of Variance ; China ; epidemiology ; Female ; Humans ; Interviews as Topic ; Multivariate Analysis ; Papillomaviridae ; Papillomavirus Infections ; epidemiology ; Risk Factors ; Sexual Behavior ; Sexual Partners ; Surveys and Questionnaires ; Tumor Virus Infections ; epidemiology ; Uterine Cervical Neoplasms ; epidemiology ; virology

OBJECTIVETo evaluate the prognosis of cervical intraepithelial neoplasia grade 1 (CIN1) at different follow-up time points in Chinese women and the relationship with high-risk human papillomavirus (HR-HPV) infection.

METHODSBiopsy-confirmed CIN1 women were followed up from cervical cancer screening cohorts established during 1999 to 2008 in Xiangyuan county, Yangcheng county, Qinxian county and Wuxiang county, Shanxi Province.In each follow-up visit, participants were examined by visual inspection with acetic acid, liquid-based cytology and HR-HPV DNA testing. Those with any positive results received colposcope and biopsies. The cumulative incidence rates of CIN grade 2 or worse (CIN2+) and CIN grade 3 or worse (CIN3+), regression rates and persistent rates were calculated using pathological findings as a gold standard. The risks of progression related with HR-HPV were evaluated stratified by baseline and follow-up HR-HPV status.

RESULTSA total of 228, 224, 261 and 105 CIN1 women received the 1-year, 2-year, 6-year and 11-year follow-up exams, respectively. The cumulative incidence rate of CIN2+ among baseline HR-HPV positive women was 4.8% (6/126), 10.7% (16/150), 16.9% (29/172) and 35% (19/55) in the above follow-up visits, respectively, and their risk of progression was 2.7(95%CI:0.3-22.0), 2.9 (95%CI:0.7-12.1), 12.0 (95%CI:1.7-86.2) and 30.6 (95%CI:1.9-493.5) times higher than baseline HR-HPV negative women. Moreover, the cumulative incidence of CIN2+ among women with positive HR-HPV both at baseline and follow-up visit was 11% (6/55), 14% (6/42), 17% (10/60) and 50% (13/26) in the above follow-up visits, respectively.No new CIN2+ cases were found among those with negative HR-HPV both at baseline and follow-up visits.

CONCLUSIONGiven that CIN1 progression is related to HR-HPV infection, different follow-up intervals and strategies for CIN1 should be taken according to HR-HPV infection status.

Aged ; Biopsy ; Cervical Intraepithelial Neoplasia ; Disease Progression ; Female ; Humans ; Papillomavirus Infections ; Prognosis ; Prospective Studies ; Uterine Cervical Neoplasms

Objective:To evaluate the calf muscle cross-sectional area (CSA) and fat fraction (FF) in amateur marathon runners based on T 2WI-Flex fat-suppression sequence and iterative decomposition of water and fat with echo asymmetrical and least-squares estimation quantitation sequence (IDEAL-IQ) technique, and to explore the correlation between them and age, body mass index (BMI), running age, speed exercise, and monthly exercise. Methods:From September to November 2022, 37 amateur marathon group (marathon group) and 12 healthy volunteers (control group) who did not often exercise were recruited in Shijiazhuang. T 2WI-Flex fat-suppression sequence and IDEAL-IQ sequence were performed on all subjects. The anterior group of the right calf (tibialis anterior, extensor hallucis longus, extensor digital longus), external group of the calf (peroneus longus, peroneus brevis), soleus, medial head and lateral head of the gastrocnemius were delineated along the edge of the muscle contour as regions of interest, and the CSA and FF values of the muscles were obtained. Twelve subjects matched with the clinical data of the control group were selected from the marathon group (marathon matched group), and the differences in CSA and FF were compared by independent sample t test or Mann-Whitney U test. Spearman correlation analysis was used to evaluate the correlation between CSA and FF values of the right calf and running-related indicators including age, BMI, running age, speed exercise, and monthly exercise in 37 amateur marathon runners. Results:The CSA of the right calf soleus muscle in the marathon matched matched group was larger than that in the control group ( t=-2.09, P=0.048). There was no significant difference in CSA of other calf muscles between the two groups ( P>0.05). The FF values of the right calf anterior group, the outer group, the soleus muscle, the medial head and the lateral head of the gastrocnemius muscle in the marathon matched group were lower than those in the control group ( P<0.05). The BMI of 37 amateur marathon runners was positively correlated with the CSA and FF values of the anterior group, the outer group, the soleus, the medial head and the lateral head of the gastrocnemius muscle (CSA: r=0.628, 0.468, 0.680, 0.566, 0.615, respectively, all P<0.05; FF value: r=0.395, 0.567, 0.631, 0.482, 0.516, respectively, all P<0.05). The FF values of the anterior group, the outer group, the soleus, the medial head and the lateral head of the gastrocnemius muscle were negatively correlated with monthly exercise ( r=-0.337, -0.405, -0.437, -0.338, -0.446, respectively, all P<0.05). Conclusions:Long-term running training can reduce the FF value of each calf muscle group, and the FF value of calf muscles is negatively correlated with the amount of running each month. Long-term running training has the greatest effect on the CSA of soleus muscle.

OBJECTIVE：To stud y the pharmacok inetics of PELGE-crebanine nanopartic les （PELGE-Cre-NPs） in rabbits. METHODS：Totally 6 rabbits were collected ，and injected with PELGE-Cre-NPs （3.5 mg/kg）via ear vein. 1 mL of blood samples were collected at 5，15，30，60，90，120，150，180，240，300 min after administration from the ear vein. After the plasma were isolated and Cre were extracted with ethyl acetate ，HPLC method was adopted to determine the plasma concentration of Cre by using verapamil hydrochloride as internal standard. The plasma concentration-time curve was drawed and pharmacokinetic parameters were calculated by using DAS 2.0 software. Chromatographic conditions such as the chromatographic column was Agilent ZORBAX Extend-C 18；the mobile phase consisted of methanol- 0.01％ triethylamine solution （75 ∶ 25，V/V）；the flow rate was 1 mL/min；the detection wavelength was 280 nm；the column temperature was 30 ℃；the injection volume was 20 μL. RESULTS：The linear range of Cre were 45.0-3 600 µg/L（R2＝0.999 9）. RSDs of inter-day and intra-day precision and stability tests were all lower than 5％（n＝6 or n＝12）；the accuracies were （97.44±2.41）％-（98.45±3.87）％（n＝6）. PELGE-Cre-NPs was in a two-compartment model in rabbits. Main pharmacokinetic parameters included that t1/2 was（109.357±33.917）min；CL was（0.016±0.001）L/（min·kg）；MRT was （76.733±7.502）min；cmax was（3 699.458±287.713）μg/L. CONCLUSIONS：The half-life period of PELGE-Cre-NPs in rabbits is longer than that of Cre injection；its retention time in the body is prolonged ，and sustained-release effect is obvious.

Left ventricular aneurysm (LVA) is a common complication of myocardial infarction. Traditional medical and surgical treatments are not effective or require high doctors' operational skills and patients' physical fitness. With the development of minimally invasive medical devices, it becomes possible for revivent TC system to treat LVA and reconstruct the left ventricle. This study introduces an existing product and its defect when used. From the perspective of clinical needs, we propose a new design of revivent TC system which realizes accurate force measurement and simplifies surgery.
Cardiac Surgical Procedures ; Heart Aneurysm ; Heart Ventricles ; Humans ; Myocardial Infarction

AIM: To investigate the protective effect of "DuZhong-DangGui" (DZ-DG) on the knee tissue of rats with osteoarthritis (OA) and its regulation role on NLPR3 inflammasome. METHODS: Twenty-four SPF male SD rats, eighteen rats were randomly selected to establish OA model by anteri- or cruciate ligament amputation (ACLT) method, and rats were divided into control group, OA model group, DZ-DG group and positive drug group, 6 in each group, treatment for 8 weeks. The peripheral blood were collected, ELISA was used to detect the levels of IL-1β, IL-6, IL-18, and TNF-α; cartilage tissue of knee joint were collected, HE staining was used to observe pathology changes, OARSI staining was used to observe cartilage calcification and preform quantitative OARSI scoring; immunohistochemistry and TUNEL staining were used to detect the contents of Caspase1 and Collagen II and the number of apoptosis in the tissue, respectively, and western blot was used to detect the protein expressions of matrix metalloproteinase-13 (MMP-13), tissue inhibitor of metalloproteinase-1 (TIMP-1), p53, p21, NLPR3, apoptosis-associated speck-like protein containing a CARD (ASC) and Pro-Caspase-1. RESULTS: Compared to control group, OA model group rats serum levels of IL-1β, IL-6, IL-18, and TNF - α significantly increased (P<0.01), OARSI scores significantly increased (P<0.01), chondrocyte apoptosis was increased (P<0.01), Caspase-1 content and MMP-13, p53, p21, NLPR3, ASC, Pro-Caspase-1 protein expressions significantly increased (P <0.01), while Collagen II content and TIMP-1 protein expression significantly decreased (P<0.01). Compared with the OA model group, DZ-DG group and positive drug group rats serum levels of IL-1β, IL-6, IL -18 and TNF - α significantly decreased (P< 0.05), chondrocyte apoptosis were significantly decreased (P<0.01), Caspase1 content, MMP-13, p53, NLPR3, Pro-Caspase-1 significantly decreased (P< 0.05), Collagen Ⅱ content and TIMP- 1 protein expression (P<0.01); DZ-DG group rats protein expression of p21 and ASC were decreased (P<0.01). CONCLUSION: The DZ-DG have protection role on cartilage of OA rats, its effect may related to mediation of NLPR3/ASC/Pro-Caspase-1 pathway, to decrease of IL-1β, and inhibition of cell apoptosis.

Objective:To investigate the effects of caspase-11 non-canonical inflammasome on the Leptospira interrogans ( L. interrogans)-induced secretion of inflammatory cytokines in J774A.1 cells. Methods:Murine mononuclear macrophage cells (J774A.1) were infected with L. interrogans strain 56601. Expression of caspase-11, IL-1β, IL-1α and IL-18 at mRNA level in J774A.1 cells were detected by real-time RT-PCR. The levels of caspase-11, IL-1β, IL-1α and IL-18 in the culture supernatants of J774A.1 cells were detected by ELISA. Results:Real-time RT-PCR showed that caspase-11 expression at mRNA level was 5.12, 14.21, 8.94, 14.06, 18.58 and 0.93 times of that in uninfected cells after 1, 2, 4, 8, 12 and 24 h of L. interrogans infection, and respectively decreased to 0.10, 0.07, 0.10, 0.09, 0.07 and 0.45 times after caspase-11 inhibitor intervention ( P<0.05). Expression of IL-1β, IL-1α and IL-18 at mRNA level was significantly increased after infection ( P<0.05). After the intervention with caspase-11 inhibitor, IL-1β mRNA decreased to 0.05, 0.03, 0.02, 0.05, 0.06 and 0.02 times ( P<0.05); IL-1α mRNA decreased to 0.14, 0.07, 0.15, 0.10, 0.03 and 0.06 times ( P<0.05); IL-18 mRNA decreased to 0.08, 0.10, 0.16, 0.18, 0.10 and 0.07 times ( P<0.05). ELISA results showed that the expression of caspase-11, IL-1β, IL-1α and IL-18 at protein level was significantly increased. After the intervention with caspase-11 inhibitor, caspase-11 level decreased to 43.07, 41.64, 51.96, 86.56, 105.36, and 129.95 pg/ml ( P<0.05); IL-1β level decreased to 15.01, 14.19, 68.02, 31.20, 173.13 and 104.98 pg/ml ( P<0.05); IL-1α level decreased to 12.14, 15.40, 38.01, 21.97, 24.48 and 27.09 pg/ml ( P<0.05); IL-18 level decreased to 96.27, 102.21, 85.34, 116.28, 155.36 and 114.03 pg/ml ( P<0.05). Conclusions:Caspase-11 non-canonical inflammasome was involved in the mediation of IL-1β, IL-1α and IL-18 secretion in mouse mononuclear macrophages after L. interrogans infection.