OBJECTIVE: Embryonic Stem Cell (ES) are characterized by totipotency and normal karyotype and provide the theoretical base in the following fields: embryonic developmeut of mammalian, cell differentiation, expres sion of exogenous genes and utilization of Ess to develop animal model for human inherited diseases. The identification of ES therefore is very important to research and utilize ES.DATA SOURCES: The relevant articles to embryonic stem cell between January 1980 and December 2003 were computer searched for in Medline with the key words "embryonic stem cell, embryo, Alkaline phosphatase,Oct-4"in English. Similarly, the relevant articles to embryonic stem cell between January 1980 and December 2003 were computer searched for in China Journal Full-text Database (CJFD) with the key word "embryonic stem cell" in Chinese.STUDY SELECTION: The articles were browsed firstly. The inclusion criteria were for those articles about the identification of embryonic stem cells. The exclusion criteria were for those about repetitive studies, reviews and other similar articles.DATA EXTRACTION: 16 articles about the identification of embryonic stem cells were collected. Then, the full-texts of the articles were looked through.DATA SYNTHESIS: The selected data were summarized in the following order: ①Preliminary identification of Ess based on morphology and growth;②Immunological valuation; ③Chromosome related identification; ④Identi fication of totipotency and pluripotency.CONCLUSION: The identification of embryonic stem cells is not the result of only one identifying method, but a process of identification. During this process of comprehensive identification, it is recommend to conduct AKP test firstly, karyotype analysis secondly, then examination of surface markers and finally identification of Ess totipotency when Ess are sufficient, takingcare to repeat every identification.

Objective To investigate the expressions of cell phenotypes CD_8 and CD_(28) of peripheral blood lymphocytes in patients with colorectal cancer. Method CD_8 and CD_(28) of cell phenotypes were measured by flow cytometry in 50 patients with colorectal cancer (cancer group) and 30 nontumorous patients (control group). Results The expression of CD_8 in cancer group was significantly higher than that in control group [(32.24±8.38)% vs (22.18±7.55)%](P < 0.01). CD_(28) and CD_8~+/CD_(28)~+ were much lower than those in control group [(52.03±10.94)% vs (60.60±7.98)%,12.18±4.28 vs 16.38±4.94](P<0.01). CD_8~+/CD_(28)~+ in Dukes D stage patients were significantly lower than that in Dukes B stage patients (P<0.05). CD_(28) and CD_8~+/CD_(28)~+ in lymph node metastasis patients were much lower than those in lymph node negative patients (P < 0.01). CD_(28) and CD_8~+/CD_(28)~+ in serosa involved patients were lower than those in no serosa involved patients (P < 0.01). Conclusions The expressions of cell phenotypes CD_8 and CD_(28) of peripheral blood lymphocytes in patients with colorectal cancer are closely related to biological characteristics of the tumor. The assays of cell phenotypes CD_8 and CD_(28) might be useful for evaluating the immunal statement and prognosis of patients with colorectal cancer.

Objective To study the relationship among the genome mutation,viral protein structures and virus evolution of human Bocavirus 2(HBoV2).Methods A method of PCR was applied to detect HBoV2 genome in 205 fecal specimens collected from children who had diarrhea in Ruian region,Zhejiang province,China.HBoV2-positive specimens were chosen to further amplify the HBoV2 genome.All the tertiary structures were predicted using the Swiss-PdbViewer software,and a phylogenetic analysis was performed by the MEGA software (version 5.1).Result The positive rate of HBoV2 PCR detection in the collected 205 fecal specimens was 5.36％ (11/205).We obtained a sequence of a nearly full-length HBoV2 genome named as Ruian isolate,which is 5196 bases in length.Alignment of the Ruian HBoV2 sequence with other HBoV2 sequences in GenBank revealed that the new HBoV2 shared 99.79％ similarities with the reference genome (GenBank no NC_012042).No mutation was found in the NP1 gene,whereas sense mutation of NS1,VP1 and VP2 gene were found.Residue solvent accessibilities that relates to hydrophobic interactions were changed between the NS1 of the Ruian isolate and the reference isolate.The protein structure and location of H-bonds were different between the NS1 of the Ruian isolate and the reference isolate.The phylogenetic tree shows a clear division of the NS1,which has a potential evolution toward or a closely evolutionary relationship with the NS1 of HBoV4.Conclusion HBoV2 infection was detected in Ruian region,Zhejiang province.We found sense mutations in the HBoV2 genome of Ruian isolate.These mutations may alter HBoV2 protein structures,and may have an evolutionary significance.These findings provide data of HBoV2 genome mutation.

Objective To investigate the expressions of homeobox gene 10 (HOXD10) and analyze its clinical significance. Methods Expressions of HOXD10 protein was detected by immunohistochemistry SP method in 53 cases of colorectal carcinoma tissues and corresponding normal tissues which was fixed by 4% formalin and embedded by paraffin.It was analyzed that the relationship between the expression of HOXD10 protein and clinico-pathological features. Results The positive staining rate of HOXD10 protein in normal colorectal mucosal tissue (5.7%)was significantly lower than that incolorectal carcinoma tissue(64.2%),the difference was statistically sig-nificant(P<0.05). In colorectal cancer tissue,the positive rate of HOXD10 protein in high differentiation(53.8%), T1+T2(38.5%),Ⅰ+Ⅱ(54.3%)and no lymph node metastasis(55.3%)was lower than that in low differentiation (73.0%),T3+T4(72.5%),Ⅲ+Ⅳ(83.3%)and lymph node metastasis,the difference was statistically significant (P < 0.05). However,it was not statistically significant between the positive rate of HOXD10 protein and the gender,age,primary site and tumor size in colorectal cancer patients(P>0.05). Conclusion The expression of HOXD10 protein is closely related to the invasion and metastasis of colorectal cancer.

Objective: To investigate the expression level of miR-9 in esophageal cancer and its effect on the biological function of esophageal cancer cells. Methods: The expression levels of miR-9 and Golgi phosphoprotein 3 (GOLPH3) in esophageal cancer and its adjacent tissues were detected by real-time fluorescence quantitative PCR (qRT-PCR). The miR-9 mimics was transfected into esophageal cancer EC109 cells, and the expression level of miR-9 was detected by qRT-PCR. The effects of overexpression of miR-9 on the biological function of EC109 cells were determined by the MTT assay, plate colony formation assay, Transwell migration assay and flow cytometry. The wild and mutant GOLPH3 double luciferase reporter gene vectors were constructed, and luciferase activity was detected. The effects of overexpression of miR-9 on the expression levels of GOLPH3 mRNA and protein were detected by qRT-PCR and Western blot. Results: Compared with the adjacent tissues, the expression level of miR-9 in esophageal cancer tissues decreased significantly (P＜0.01), while that of GOLPH3 increased significantly (P＜0.01). Compared with the negative control group, the expression level of miR-9 in EC109 cells transfected with miR-9 mimics increased significantly (P＜0.01), and the proliferation and migration ability of the EC109 cells decreased obviously (P＜0.01). The cell cycle of the EC109 cells was blocked in G2/M phase (P＜0.01). The dual luciferase reporter assay, qRT-PCR and Western blot confirmed that miR-9 could bind with GOLPH3 specifically (P＜0.01), and mediate the degradation of GOLPH3 mRNA (P＜0.01), which led to the decrease of GOLPH3 protein expression level (P＜0.05). Conclusion MiR-9 is low expression in esophageal cancer, and may participate in the occurrence and development of esophageal cancer by regulating GOLPH3.

To evaluate the safety and efficacy of oral double-balloon enteroscopy for small intestinal bezoars with obstruction, the clinical, endoscopic, imaging, pathological and surgical data of patients with small intestinal bezoars and obstruction treated in Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from December 2021 to March 2023 were retrospectively analyzed. A total of 5 patients, all female, aged 52-83 years, were included. The clinical manifestation was mainly digestive tract obstruction. The diameter of bezoars was 30-50 mm. Three were located in the jejunum, one in the proximal ileum, and one at the jejuno-ileal junction. Three cases were diagnosed as having small intestinal bezoars by imaging examination before operation. All 5 patients achieved technical success. Four patients achieved clinical success, the symptoms of obstruction were completely relieved, and 1 patient was partially relieved due to malignant obstruction. No obvious complication occurred in 5 patients after operation. The follow-up time was 15-790 days, and there was no recurrence. The results showed that oral double-balloon enteroscopy for small intestinal bezoars was safe and effective, providing a new treatment for patients with small intestinal bezoars except for surgery.