Objective:To evaluate the value of CK19 mRNA detection in diagnosis of breast cancer. Methods: Patients were categorized into 3 groups:Group 1 consisting of 77 stageⅠ/Ⅱbreast cancer patients, group 2 consisting of 77 stage Ⅲ/Ⅳ breast cancer patients, and control group consisting of 40 non-cancer volunteers. Total RNA was extracted from peripheral blood cells with the RNA Kit as described by the manufacturer, which was followed by one step RT-PCR of RNA. Agarose gel electrophoresis and sequencing were carried out to confirm the amplification of expected sequence (314 bp). ?~2 test was used to compare CK19 mRNA positive rates in different stages of breast cancer. Results: 15/77 (19.48%) of stage Ⅰ/Ⅱ breast cancers patients, 31/77 (40.26%) of stage Ⅲ/Ⅳ breast cancers patients and 2/40 (5.00%) of non-cancer volunteers demonstrated positive RT-PCR results. The positive rate of CK19 mRNA in peripheral blood of stage Ⅰ/Ⅱ or stage Ⅲ/Ⅳ breast cancer patients was significantly higher than that of non-cancer volunteers(P=0.037, P=0.001). The positive rate of CK19 mRNA in peripheral blood of stage Ⅲ/Ⅳbreast cancer patients was significantly higher than that of stageⅠ/Ⅱbreast cancer patients(P=0.005). Conclusion: CK19 RT-PCR is a promising method for detecting circulating tumor cells in patients with breast cancer, which may provide valuable information for tumor staging and treatment.

Objective To analyze the surgical management and prognosis of hilar cholangiocarcinoma(HCC).MethodClinical data of 72 patients with HCC were retrospectively analyzed.Results There was a significant difference in the 3- and 5- year survival rates and median survivals among the groups treated with different surgical methods(x2 =28.1,P＜ 0.01 ).Univariate analyses conducted for the 16 factors which might affect prognosis showed the surgical method,lymphatic metastasis,positive margin,as well as the type of pathology and staging were of statistically significant prognostic value (P＜0.05).On a multivariate analysis using the Cox proportional hazards model,surgical management,lymphatic metastasis and the type of pathology and staging were independent prognostic factors (regression coefficient 1.4157,1.1072,1.6435).ConclusionRadical excision is an important prognostic factor for HCCa and extended resection improved the long-term survival rates of radical surgery.

Objective: To explore childhood traumatic experiences of OCD patients who were also diagnosed personality disorders (PDs) and those without PDs. Methods:SCID-I/P was administrated to diagnose OCD patients. PDQ+4 and PDI-IV were then administrated to 44 OCD subjects to determine whether they also suffered from personality disorders. Childhood maltreatment experiences of the sample were measured with Childhood Trauma Questionnaire (CTQ).50 normal control were also collected and subjected to CTQ-SF tests. Results:32 of 44 (72%) OCD patients also met DSM-IV criteria for personality disorders. OCD patients scored significantly higher than normal control in CTQ-SF (Childhood Truma Questionuaire-28 Short Form) and in all domains with the exception of physical abuse. Comparison between OCD subjects with PDs and normal control, the former scored significantly higher in emotional abuse (55.10/32.54), sex abuse (49.63/36.30), physical neglect (52.03/34.76), and emotional neglect domains (55.84/32.32). Whereas OCD subjects without PDs only scored significantly higher in emotional neglect domains compared with normal control (42.50/28.26).Conclusions:Subjects with PDs experienced more severe traumatic events during their early life. "pure OCD" patients, however, might experienced more emotional neglect.

Objective To establish a method for quantitative detection of hepatitis B virus covalently closed circular DNA(HBV cccDNA ) in infected cells. Methods The transfected cell line HepG2.2.15 which can consistently produce Dane particles was maintained in DMEM containing 380 ?g/ml G418 and 10% fetal bovine serum. Cells in the exponential period were harvested from flasks, then intracellular HBV cccDNA was extracted from pellet containing 1?10~6 cells with mini plasmid extraction kit (QIAGEN).The extraction product was further purified by mung bean nuclease to remove HBV relaxed circular DNA possibly remained. HBV cccDNA was quantitatively detected by fluorescent PCR with selective primer set and Taqman MGB probe. Culture medium before exponential period, HBV DNA positive and negative serum samples from patients with chronic hepatitis B (mild) were amplified simultaneously to test the specificity of the fluorescent PCR method. Plasmids containing whole HBV genome were amplified with the same primer set and fluorescent probe to determine the sensitivity of the method. Results HBV cccDNA was detectable in HepG2.2.15, and the average quantity was 18 copies per cell approximately. No detectable fluorescent signal was observed when culture and serum samples were amplified. The detectable HBV cccDNA was as low as 10~3 copies per ml at least by this method. Conclusions This method is convenient, highly specific and highly sensitive. It can be utilized in the quantitative detection of intracellular HBV cccDNA as well as in the screening and evaluation of antiviral agents.

Objective To evaluate the feasibility and efficacy of Separate stent placement in the palliative treatment of inoperable malignant gastroduodenal obstruction.Methods Thirty four consecutive patients with inoperable malignant gastroduoenal obstruction were treated with peroral placement of Separate stent. Gastroduodenal obstruction was caused by gastric ( n =11), pancreatic ( n =12), matastatic ( n =3), biliary duct ( n =6) or duodenal ( n =2) cancers. Separate outer partial covered stent overlapping with inner bare stent were placed coaxially under fluoroscopic guidance.Results The technical successful rate reached 97% (33 of 34) with no major complication. Symptoms of gastroduodenal obstruction relieved in 33 patients. The average score of food intake capacity improved from 3.8 to 1.2 after Separate stent placment. Stent migration and tumor ingrowth were not detected during the follow up period. Tumor overgrowth was confirmed in one patient and a second stent was placed to solve the problem.Conclusions Peroral placement of Separate stent is a feasible and effective treatment for patients with inoperable malignant gastroduodenal obstruction.

OBJECTIVE To study the synchronous electromyograhic features of suprahyoid muscles [anterior digastric muscle(AD), posterior digastric muscle(PD),stylohyoid muscle(STH),mylohyoid muscle(MH)and geniohyoid muscle(GH)] and intrinsic laryngeal muscles [lateral cricoarytenoid muscle(LCA)and posterior cricoarytenoid muscle(PCA)]. METHODS Synchronous electromyograhic features of 13 dogs were observed under the circumstances of breath, swallowing and phonation. RESULTS ①AD, PD, STH, MH, GH and PCA discharged during inspiratory phase, while LCA discharged during expiratory phase. ②During swallow, suprahyoid muscles and LCA showed intensive synchronous discharge. The myoelectricity of PCA was suppressed while myopotential can be detected during intermission. ③During phonation, AD, PD, LCA and PCA showed an intensive synchronous discharge, while emergence time of PCA was later. GH and STH showed no myoelectricity activities. CONCLUSION①During phonation and swallowing, MH is synchronized with LCA. It can be chosen as the backup muscle for the reestablishment of glottic adduct function. ②During inspiratory phase, GH is synchronized with PCA. It can be chosen as the backup muscles for the reestablishment of vocal cord abduct function.

Objective:To investigate the high risk factors of pelvic lymph node metastasis in patients with type Ⅰ endometrial carcinoma in order to provide the basis for making reasonable operation scope.Methods:Risk factors of pelvic lymph node metastasis were analyzed in 136 cases of type Ⅰ endometrial carcinoma.Univariate analysis was performed with Chi square test or Fisher's exact probability method,and multivariate analysis was performed with a logistic regression mode.Results:The positive rate of pelvic lymph nodes in 136 patients with type Ⅰ endometrial carcinoma was 9.56％ (13/136).Univariate analysis showed that histological grade,size of lesion,depth of myometrial invasion and vascular invasion were related to lymph node metastasis(P ＜0.05);Multivariate Logistic analysis showed that low differentiation,deep muscular invasion,tumor diameter≥2 cm and LVSI were independent risk factors of pelvic lymph node metastasis in patients with type Ⅰ endometrial carcinoma(P ＜0.05).Conclusions:The rate of pelvic lymph node metastasis is low in type Ⅰ endometrial carcinoma.Patients with low differentiation,deep muscular invasion,tumor diameter≥2 cm and LVSI are more likely to occur pelvic lymph node metastasis in type Ⅰ endometrial carcinoma.

Abstract Hypertensive disorders of pregnancy (HDP) are a common severe complication during pregnancy, which is characterized by complex etiology, unclear pathogenesis and lack of effective tools for early diagnosis and prediction. Recently, the development of omics technology provides new insights into the research into HDP. Based on national and international publications from 2011 to 2022, this review summarizes the application of genomics, proteomics, metabolomics, and microbiomics in the pathogenesis and prediction of HDP, so as to provide insights into the prediction, prevention and precise treatment of HDP.

Objective To determine whether hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) could be detected in serum of patients with hepatitis B and evaluate the related factors and clinical significances. Methods Fifty-seven patients, including 26 with mild chronic hepatitis B (CHB) and 31 with severe hepatitis B (SHB) were enrolled. Prothrombin time (PT), hepatic biochemical indexes, serum markers of hepatitis virus, serum total HBV DNA and HBV cccDNA of every patient were detected after hospitalization. Factors associated with the detection rate of serum HBV cccDNA were analyzed using Logistic stepwise regression. Results Serum HBV cccDNA was detected in 13 patients with SHB and 1 with mild CHB, and serum levels of HBV cccDNA were varying from 1.25 × 103 to 4. 88 × 104 copy/mL. The detection rates were significantly different between the two groups (P=0. 0014). The sensitivity and specificity of SHB diagnose by serum HBV cccDNA detection were 41.94% and 96.15 %, respectively. Logistic regression analysis showed that the detection rate of serum HBV cccDNA was associated with PT (X2 = 7. 2192, P= 0. 0072), while not associated with age, sex, total serum HBV DNA, total bilirubin or alanine aminotransferase (ALT). Conclusion Serum HBV cccDNA could be detected in some of the patients with SHB, whic hmay be considered as one of the diagnostic indexes for SHB,

Objective:To investigate the prevalence and related factors of dental caries in 3-year-old children in the Development District in Dalian.Methods:A multi-stage,randomized,cluster sampling method was used in the cross-sectional study.All 3-year-old children in the selected 15 kindergartens were invited to take part in the oral health examination conducted by 3 calibrated examiners.The WHO recommended caries diagnosis criteria were used.The structured questionnaire including the information of children's oral hygiene habits,dietary habits,parents' oral health knowledge and socioeconomic status was completed by the parents.Results:1 220 3-year-old children completed the clinical oral examination and 964 children's parents(79.0％) finished the questionnaire.The caries prevalence in 3-year-old children was 62.8％,and the mean dmft and dmfs scores were 3.40 and 7.34,respectively.No differences in the prevalence of caries,mean dmft and dmfs scores were found between boys and girls(P ＞0.05).Logistic regression analysis showed that the frequency of sweets and sugar-containing beverages consumption(P＜0.05),parents'education level(P＜0.05) were significantly related to the caries prevalence.Conclusion:The prevalence and severity of dental caries in 3-year-old children in Dalian Development District were high and the frequency of consumption sugar-containing foods and beverages,parents'education level were related to the decay prevalence.