Objective To observe the expression of P21 and CDK6 in cervical squamous cell carcinoma,and to investigate the relationship between their expression and cervical squamous cell carcinoma.Methods The expression of CDK6 and P21 in 100 cases of cervical squamous cell carcinoma,20 cases of cervical CIN lesions and 20 cases of normal cervical tissues were detected by immunohistochemical ABC method,and the relationship between them and tumor differentiation,invasion depth,lymph node metastasis and clinical stage were analyzed.Results The expression rates of P21 and CDK6 in cervical squamous cell carcinoma tissues were significantly higher than those in CIN lesions and normal cervical tissues;The low expression of P21 was associated with the depth of tumor invasion,lymph node metastasis and clinical stage (P＜0.05),and was not associated with the degree of tumor differentiation (P＞0.05);The high expression of CDK6 was associated with tumor differentiation,invasion depth,lymph node metastasis and clinical stage (P＜0.05).Conclusion The abnormal expression of P21 and CDK6 may play an important role in the occurrence and development of cervical squamous cell carcinoma,and the two may have a certain significance in the prognosis of cervical squamous cell carcinoma.

Objective To survey the current situation of fertility intention to have a third child among childbearing age women in Wanzhou District of Chongqing Municipality under the background of the new fer-tility policy,and to analyze its influencing factors.Methods The convenience sampling method was adopted to conduct the survey on the third child fertility intention by the self-designed questionnaire and the Question-naire Star platform among the childbearing women with one child or two children in Wanzhou District of Chongqing Municipality from September to October 2022.The multiple linear regression analysis and two-step cluster analysis were used to explore the influencing factors.Results A total of 632 questionnaires were re-ceived,32 invalid questionnaires were excluded,and the effective recovery rate was 94.9％.The score of fertili-ty intention to have a third child in the respondents was(12.81±2.83)points.The multiple linear regression analysis showed that the age,marital status,place of residence,spouse cultural degree,family rearing child model,whether the family can provide support and previous vaginal delivery history were the influencing fac-tors of fertility intention to have a third child among the childbearing age women in this place(P＜0.05).The two step cluster analysis showed that"obstetrician/midwife's help"was the most important predictive factor for the childbearing age women to choose to have a third child.Conclusion The current situation of childbear-ing age women's fertility intention to have a third child in Wanzhou District of Chongqing Municipality is not optimistic.It is necessary to create the positive factors that encourage childbearing age women to make deci-sion about having a third child.

Objective:To investigate the current status and influencing factors of kinesiophobia in adult burn patients.Methods:A single center cross-sectional research method was conducted. A total of 170 adult burn patients, meeting the inclusion criteria, were admitted to the Department of Plastic Surgery and Burns of the West China Hospital of Sichuan University from October 2018 to December 2019. On admission, the self-made general information questionnaire was used to investigate the gender, age, education level, marital status, payment method of medical expenses, injury factors, and total burn area of patients. One month after admission or before discharge, the presence and degree of kinesiophobia of patients were evaluated by the Tampa Scale for Kinesiophobia (TSK), their pain degrees (results averaged) at the time of burn, debridement and dressing change, after burn operation, and at rest were evaluated by the Visual Analogue Scale, their social support levels were evaluated by the Social Support Revalued Scale, and their degrees of anxiety and depression were evaluated by the Hospital Anxiety and Depression Scale. According to the TSK score, the patients with score >37 points were included into kinesiophobia group, and the patients with score ≤37 points were included into non-kinesiophobia group. The general information of patients in the two groups, as well as the pain score, social support level score, anxiety score, and depression score mentioned above, were recorded. Data of patients between the two groups were statistically analyzed with unifactor analysis including chi-square test, independent sample t test, and Mann-Whitney U test. The factors with statistically significant differences in unifactor analysis were used as variables for multivariate logistic regression analysis to screen out the independent influencing factors of kinesiophobia in adult burn patients. Results:Questionnaires and scales of 170 patients were collected, and the recovery rate was 100%. The data of two patients complicated with cranial fracture aggravation were removed, and 168 valid data were obtained, with the effective rate of 98.82%. Among the 168 patients, 88 were male (52.38%) and 80 were female (47.62%), aged from 18 to 71 (41±6) years. Most of the patients had secondary school education or below, were married, and with no out-of-pocket medical expenses. The main factors of injury were flame and hydrothermal fluid, and the total burn area was 2%-75% ((28±5)%) total body surface area. The TSK score was (41±5) points, the pain score was 4.0 (2.6, 7.0) points, the social support level score was (40±5) points, the anxiety score was 8.5 (7.0, 13.0) points, and the depression score was 9.5 (6.5, 12.0) points. A total of 98 patients had kinesiophobia, and the incidence of kinesiophobia was 58.33%. There were no statistically significant differences in gender, age, educational level, marital status, or injury factors of patients between kinesiophobia group and non-kinesiophobia group ( P>0.05). The percentage of out-of-pocket expenses, total burn area, pain score, anxiety score, and depression score of patients in kinesiophobia group were significantly higher than those in non-kinesiophobia group ( χ2=6.402, t=2.39, Z=-8.05, -6.68, -7.89, P<0.05 or P<0.01), and the social support level score of patients in kinesiophobia group was significantly lower than that in non-kinesiophobia group ( t=5.22, P<0.01). The multivariate logistic regression analysis showed that total burn area, pain score, social support level score, anxiety score, and depression score were the independent influencing factors for the development of kinesiophobia in adult burn patients (odds ratio=0.79, 1.45, 0.78, 1.15, 1.17, 95% confidence interval=0.80-0.92, 1.24-1.74, 0.65-0.91, 1.06-1.29, 1.03-1.24, P<0.01). Conclusions:The incidence of kinesiophobia in adult burn patients is high, and the degree of kinesiophobia is mainly affected by total burn area, pain, social support level, anxiety and depression degrees, and so on. Thus these factors should be taken into consideration when designing interventions to reduce the incidence and degree of kinesiophobia.

OBJECTIVETo construct the specific stable expression and high efficiency small interfering RNA(siRNA) expression vector that can block DNMT1 gene function.

METHODSUsing vector-based RNA interference technique, the authors constructed a vector to transcribe functional short interfering RNA (RNAi). After transfection by lipofectmine (TM) reagent, the treated cells were selected by G418. The expression levels of RNA and protein of DNMT1 were analyzed by reverse transcription polymerase chain reaction(RT-PCR) and Western blotting. The status of methylation of E-cadherin was analyzed by methylation-specific PCR(MSP).

RESULTSThe expression level of endogenous DNMT1 mRNA in transfected SMMC-7721 cell lines with DNMT1 RNAi construct was 43% less than that in control cell 7721-pSU cell lines. The protein level in the former was about 10% less than that in the latter. The efficiency of the siRNA of DNMT1 was found to be higher than 90%. Demethylation of promoter of E-cadherin was obtained due to the inhibition of DNMT1.

CONCLUSIONDNMT1 siRNA stable expressing vector was obtained by gene-recombined technology. There was no complete sameness between the levels of protein and RNA in gene silenced cell lines. The efficiency of the siRNA should be confirmed by Western-blotting.

Blotting, Western ; Cell Line, Tumor ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; metabolism ; DNA Methylation ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; RNA Interference ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection