Biochemistry is an essential course for medical students,which updates fast,develops quickly and has strong characteristic of experimentation.Opening experiment is a new open type experimental teaching mode and students are regarded as main body.Opening experiment is helpful to stimulating activeness and cultivating innovative consciousness of students as an effective supplement for traditional experimental teaching.This kind of teaching mode is in conformance with experimental teaching reform in university and is an effective way of cultivating medical science talented men.

Objective:To investigate the role of TSLP and NF-κB in the pathogenesis of adenomyosis.Methods:Immunohisto-chemistry was employed to detect the expression of TSLP and NF-κB p65 in the endometrial glandular cell and stromal cell of 35 adeno-myosis patients and 20 hysteromyoma patients as control , and analyze the correlation of the two proteins.The concentration of TSLP was measured by specific ELISA in the serum of healthy subjects besides of the patients with adenomyosis and hysteromyoma .Results:TSLP and NF-κB p65 were both positively expressd in the glandular cells and stromal cells of ectopic and eutopic endometrium of patients with adenomyosis , their expression was significantly higher than that of the control group ( P<0.01 ) , and the expression of TSLP and NF-κB p65 in ectopic endometrium was significantly higher than that in eutopic endometrium ( P<0.01 ).There was positive correlation between TSLP and NF-κB p65 in ectopic endometrium.The concentration of TSLP was higher in the serum of patients with adenomyosis.Conclusion:The TSLP is highly expressed in endometrial glandular and stromal cells of patients with adenomyosis , and there is higher level of TSLP in serum , and there is correlation between the expression of TSLP and NF-κB in ectopic endometrium.

BACKGROUND: Many operations for isolating, purifying and identifying bone marrow mesenchymal stem calls (BMSCs) are complicated and cost much. Also they have great effect on cell activity. Whether whole bone marrow adherent culture can avoid above-mentioned disadvantages remains unclear. At present, many studies huve been done to confirm an effective and low cost method for isolating, purifying and identifying such cells.OBJECTIVE: This study is to in vitro induce and differentiate rat BMSCs by whole bone marrow adherent culture,and to identify the cells.DESIGN: A controlled observational experiment.SETTING: Qingdao University Medical College.MATERIALS: This study was carried out in the Laboratory of Oral Cavity and Laboratory of Molecular Biology (provincial level) Qingdao University Medical College between November 2005 and March 2007. Twenty Wistar rats of either gender, aged 3 to 4 weeks, of SPF grade, weighing 120-150 g, were provided by the Qingdao Laboratory Center. The protocol was carried out in accordance with animal ethics guidelines for the use and care of animals. Fetal bovine serum (FBS, Hangzhou Sijiqing Bioengineering Material Research Institute), alkaline phosphatase (ALP) kit (Nanjing Jiancheng Bioengineering Research Institute), reverse transcription kit (American Promega Corporation) and primer (Shanghai Bioengineering Co.,Ltd.) were used in this study.METHODS: Adult rat BMSCs were isolated and cultured by whole bone marrow adherent culture. They were digested with 2.5 g/L trypse and inoculated at a density of 5 ×107 L-1 in 6-well culture plate. Then, the cells were divided into experimental group and control group. Inducing culture medium was added to experimental group, and the same amount of basic culture medium was added to control group. ① Cell differentiation and calcium tuberculation were observed under the inverted microscope. ② Biological characteristics of induced cells were detected by calcium tubercle Von Kossa and alizarin Bordeaux. ③ALP activity was detected by diazo salt staining. ④Human core binding factor alpha subunit-1 (Cbf α-1), osteocalcin (OCN) and osteoblast-specific Osterix (OSX) mRNA expressions were detected by reverse transcription-polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: ① Induction and differentiation results of cells. ② Biological characteristics of cells induced by rat BMSCs. ③ ALP activity. ④ Cbf α-1, OCN and OSX expressions.RESULTS: ①Inducing culture medium was added in the serial subcultivation. About 9 days later, cell clones were connected to each other. On about 21 to 28 days, some pykno-round mineralized tubercles appeared. Meanwhile,control cells were connected to each other, but they did not form the tubercle. ② In the experimental group, when MSCs were induced for 21 to 28 days, obvious round or oval calcified tubercles were seen by naked eyes. The results of Von Kossa staining exhibited black sediments, and those of alizarin Bordeaux staining exhibited salmon tubercles. Calcium tubercles were not found in the control group. ③The ALP activity after 2 weeks of induction was obviously increased in the experimental group, but was relatively weak in the control group. ④In the experimental group,Cbf α-1, OCN and OSX expressions were significantly increased after induction.CONCLUSION: After being in vitro induced and differentiated by whole bone marrow adherent culture, rat BMSCs exhibited morphological and biological characteristics similar to typical osteoblasts.

Objective:This work aims to explore the application value of cervical exfoliated cell DNA (Cysteine dioxygenase type 1, CDO1 and CUGBP Elav-like family member 4, CELF4) methylation in the detection of endometrial cancer in women of childbearing age. Methods:From November 2021 to October 2022, a prospective study was conducted on a total number of 517 reproductive-age women with abnormal uterine bleeding who had surgical indications for hysteroscopy at the Xiangya Third Hospital of Central South University. The cervical exfoliated cells were collected for cytology, HPV (human papillomavirus) and gene methylation detection before operation. Clinical information of patients, level of tumor-related biomarkers, and endometrial thickness of transvaginal ultrasound (TVS) were also collected. Single factor regression method was used to analyze the high-risk factors of endometrial cancer. Receiver operating characteristic curve analysis was used to obtain the area under the curve(AUC), focusing on the screening efficacy of gene methylation test for endometrial cancer in women of childbearing age.Results:The age, body mass index (BMI)≥25 kg/m 2, endometrial thickness≥11 mm, CDO1 m ΔCt≤8.4, CELF4 m ΔCt≤8.8, and double gene methylation were associated with endometrial cancer in women of childbearing age, 1.16(1.08-1.25), 4.33(1.89-10.31), 9.49(3.88-26.69), 69.62(25.70-224.36), 23.64(9.66-63.99), 87.39(24.83-555.05), all P<0.05. The AUC was 0.90 (95% CI 0.83-0.97) of CDO1 m/ CELF4 m in diagnosing endometrial carcinoma was higher than others factors, with sensitivity and specificity of 91.7% (95% CI 80.6%-100%) and 88.8% (95% CI 86.0%-91.6%). TVS combined with DNA methylation detection further improved the sensitivity to 95.8% (95% CI 87.8%-100%), but could not improve the specificity 68.0% (95% CI 63.8%-72.1%). Conclusions:For women of childbearing age with abnormal uterine bleeding or abnormal vaginal discharge, the accuracy of cervical cytology DNA methyl detection of endometrial cancer is better than other non-invasive clinical programs. DNA methylation combined with TVS can improve the sensitivity of detection.

Objective: To investigate the effects and the potential mechanism of shikonin on rat random flaps. Methods: Seventy-two wistar male rats in grade SPF were randomly divided into negative control group, tetramethylpyrazine group (TMP group) or shikonin treatment group. The random skin flap sized 8 cm×2 cm, with a cephalic based pedicle, was performed on the back of the rat. Each group was administered accordingly by intraperitoneal injection once per day for 7 days: shikonin treatment group (1 mg/kg), TMP group (10 mg/kg) and control group (1 ml/kg). Morphological changes of skin flaps were observed by HE staining. The positive expression of iNOS and COX-2 in skin flap tissues after operation were detected by immunofluorescence staining. The serum contents of TNF-α and IL-6 were detected by ELISA. Results: Inflammatory cell infiltration and inflammatory reaction of flap was more severe in control group at different time points after operation. The number of inflammatory cells in shikonin treatment group and TMP group were significantly decreased, compared with controls (P<0.01). The decrease of the inflammatory cell numbers in shikonin treatment group was more significant. The proliferation of granulation tissue and fibroblast in skin flap was obvious, and the survival rates of the skin flap were significantly increased in shikonin treatment group and TMP group (both P<0.01). The numbers of iNOS or COX-2 positive cells in shikonin treatment group and TMP group were significantly decreased, when compared with controls (both P<0.01). Compared with TMP group, the numbers of iNOS and COX-2 positive cells in shikonin treatment group were significantly decreased (P<0.01 and P<0.05, respectively)in early period after operation. Compared with the control group, serum levels of TNF-α and IL-6 in shikonin treatment group and TMP group were significantly decreased (all P<0.01). The shikonin treatment group developed more significant reduce. Conclusions Shikonin can significantly improve inflammatory response of skin flap in rats, which might be related to inhibiting the expression of iNOS and COX-2, decreasing serum levels of TNF-α and IL-6 , and reducing the release of inflammatory cytokines .

As the overweight and obese population is growing， the incidence of obstructive sleep apnea is rising， and most of the cases are complicated with coronary heart disease and other cardiovascular diseases. The two diseases affect each other and seriously endanger the patients' health， becoming a major public health problem of global concern. It is of great clinical importance to explore the combination of Chinese and Western medicine in the prevention and treatment of coronary heart disease complicated with obstructive sleep apnea syndrome. Researchers have explored the relationship between the two based on traditional Chinese medicine（TCM） theory and found that the two diseases belong to the TCM disease categories of chest impediment and snoring， respectively， and their co-morbidity is associated with the abnormal physiological functions of the heart and lungs. The failure of the heart to govern blood leads to the generation of blood stasis， and that of the lung to govern Qi movement leads to the generation of phlegm. The accumulation of phlegm and blood stasis in the chest causes chest impediment and snoring due to obstruction of the airway. This paper discusses the internal linkage between the pathogenesis of coronary heart disease and obstructive sleep apnea syndrome in Chinese and Western medicine from the TCM theory of heart-lung correlation. Furthermore， this paper proposes the treatment principles of simultaneously treating the heart and lung and activating blood and resolving phlegm， aiming to provide a theoretical basis for the clinical prevention and treatment of coronary heart disease complicated with obstructive sleep apnea.