BACKGROUND: Nowadays stem cells have been isolated successfully from the epithelial tissue of human and rodents such as skin, hair follicle, cornea, oral mucous, intestinal mucosa dental epithelial cells, and salivary glands by the Chinese or foreign researchers. But according to the vaginal mucosa stem cells (VMSCs), it is not known in the field of isolation and application.OBJECTIVE: To find out the suitable method of isolation, identification, and culture of human VMSCs.DESIGN: Observational basic study. SETTING: Department of Gynaecology and Obstetrics, the Third Affiliated Hospital of Guangzhou Medical College. MATERIALS: This study was performed in the Zhongshan Ophthalmology Center of Zhongshan Medical College from October 2004 to August 2005. Two-week gestation mice were provided by the Experimental Animal Center of Guangzhou University of Chinese Medicine. The sterile human vaginal mucosa was collected from five patients (25-38 years old) after the plastic operation of vagina from the 3rd Affiliated Hospital of Guangzhou Medical College. METHODS: Vaginal mucosa cells could be harvested from human vagina mucosa treated with trypsin-collagenase digesting. The VMSCs were selected by adhesion to type Ⅳ collagen for 20 minutes. The cell cycles of the VMSCs and primary human vaginal mucosa cells were analyzed by flow cytometry. VMSCs were cultured in vitro with 4 different media, including feeder layer cells plus epidermal total culture media, feeder layer cells plus DMEM/F12 (3:1), epidermal total culture media and DMEM/F12 (3:1). At the 12th day, their colony forming efficiency (CFE), CK19 and CK10 positive rates were compared. Then VMSCs were continuously cultured and passaged till they were apoptosis.MAIN OUTCOME MEASURES: ① Positive rate of CK19 and CK10 and G0/G1 proportion; ② Positive rate of CK19 and CK10, the colony forming efficiency (CFE) and the times be passaged when primary VMSCs were cultured in different media after 12 days. RESULTS: The cells at the G0-G1 phase cell cycle, CK19 and CK10 positive rates between the VMSCs and primary human vaginal mucosa cells were statistical different (P < 0.05). When cultured at 12th day, the VMSCs cultured with feeder layer and epidermal total culture media had the highest CFE and CK19 positive rate and lowest CK10 positive rate. They could be passaged over 15 times in vitro. CONCLUSION: It is an effective way that select VMSCs by adhesion to type Ⅳ collagen for 20 minutes. In this research, culturing VMSCs in vitro with feeder layer and epidermal total culture media was the best way to keep VMSC characteristics.

Objective:To investigate the clinical application of carbon nanoparticles mapping lymph nodes in curative resection for colorectal carcinoma.Methods:Patients diagnosed with colorectal cancer before operation and undergoing radical surgery with intact postoperative pathological data in the Sixth Affiliated Hospital, Sun Yat-sen University from March 2016 to March 2018 were included in this retrospective case-control study. Those who were diagnosed with ileus, recurrent carcinoma or underwent emergency operation were excluded. A total of 1421 cases were included, with 156 cases in the carbon nanoparticles mapping group and 1265 cases in the control group. Using 1∶3 case control matching based on gender, weight, TNM staging and neoadjuvant chemotherapy, 145 and 435 cases were finally recruited in the carbon nanoparticles mapping group and control group, respectively. Patients in the carbon nanoparticles mapping group underwent preoperative colonoscopy with carbon nanoparticles submucosal injection 2.4 (1.0 - 14.0) days before operation. Carbon nanoparticles of 0.25 ml was injected at 4 points (3, 6, 9 and 12 o'clock each) 0.5-1.0 cm around the tumor. The number of eliminated lymph node, number of positive lymph node and positive rate between the two groups were compared, and the number of eliminated lymph node in different subgroups of T stage, N stage, TNM stage and neoadjuvant chemotherapy was analyzed and compared.Results:After case control matching, total number of eliminated lymph nodes in the carbon nanoparticles mapping group was significantly higher than that in the control group (22.2±11.2 vs. 19.0±9.5, t=3.025, P=0.003). However, no statistically significant differences were found in the number of positive lymph node and lymph node positive rate between two groups (all P>0.05). Subgroup analysis showed that as compared to the control group, total number of eliminated lymph nodes in the carbon nanoparticles mapping group was significantly higher in T3 stage subgroup (median: 22 vs. 18, Z=2.435, P=0.015), N0 stage subgroup (median: 20.5 vs. 17.5, Z=2.772, P=0.006), TNM II stage subgroup (median: 23.5 vs. 19.0, Z=2.654, P=0.008) and neoadjuvant chemotherapy (median: 22.5 vs. 13.0, Z=3.287, P=0.001), while compared to the control group, the number of positive lymph node (median: 4.0 vs. 6.5, Z=-2.530, P=0.011) and the lymph node metastasis degree (median: 16% vs. 31%, Z=-2.862, P=0.004) were lower in the carbon nanoparticles mapping group in N2 subgroup. Conclusion:Carbon nanoparticles mapping lymph nodes can effectively enhance the number of eliminated lymph nodes in curative resection for colorectal cancer.

Objective:To investigate the clinical application of carbon nanoparticles mapping lymph nodes in curative resection for colorectal carcinoma.Methods:Patients diagnosed with colorectal cancer before operation and undergoing radical surgery with intact postoperative pathological data in the Sixth Affiliated Hospital, Sun Yat-sen University from March 2016 to March 2018 were included in this retrospective case-control study. Those who were diagnosed with ileus, recurrent carcinoma or underwent emergency operation were excluded. A total of 1421 cases were included, with 156 cases in the carbon nanoparticles mapping group and 1265 cases in the control group. Using 1∶3 case control matching based on gender, weight, TNM staging and neoadjuvant chemotherapy, 145 and 435 cases were finally recruited in the carbon nanoparticles mapping group and control group, respectively. Patients in the carbon nanoparticles mapping group underwent preoperative colonoscopy with carbon nanoparticles submucosal injection 2.4 (1.0 - 14.0) days before operation. Carbon nanoparticles of 0.25 ml was injected at 4 points (3, 6, 9 and 12 o'clock each) 0.5-1.0 cm around the tumor. The number of eliminated lymph node, number of positive lymph node and positive rate between the two groups were compared, and the number of eliminated lymph node in different subgroups of T stage, N stage, TNM stage and neoadjuvant chemotherapy was analyzed and compared.Results:After case control matching, total number of eliminated lymph nodes in the carbon nanoparticles mapping group was significantly higher than that in the control group (22.2±11.2 vs. 19.0±9.5, t=3.025, P=0.003). However, no statistically significant differences were found in the number of positive lymph node and lymph node positive rate between two groups (all P>0.05). Subgroup analysis showed that as compared to the control group, total number of eliminated lymph nodes in the carbon nanoparticles mapping group was significantly higher in T3 stage subgroup (median: 22 vs. 18, Z=2.435, P=0.015), N0 stage subgroup (median: 20.5 vs. 17.5, Z=2.772, P=0.006), TNM II stage subgroup (median: 23.5 vs. 19.0, Z=2.654, P=0.008) and neoadjuvant chemotherapy (median: 22.5 vs. 13.0, Z=3.287, P=0.001), while compared to the control group, the number of positive lymph node (median: 4.0 vs. 6.5, Z=-2.530, P=0.011) and the lymph node metastasis degree (median: 16% vs. 31%, Z=-2.862, P=0.004) were lower in the carbon nanoparticles mapping group in N2 subgroup. Conclusion:Carbon nanoparticles mapping lymph nodes can effectively enhance the number of eliminated lymph nodes in curative resection for colorectal cancer.

【Objective】 To analyze the quality level of the laboratory pre-analytical phase, so as to take effective quality improvement interventions to further standardize the operation and provide basis for ensuring the quality of blood testing. 【Methods】 Pre-analytical phase quality indicators of blood screening laboratory in Shanghai Blood Center were established, and those had serious impact on blood safety were defined as the sentinel indicators. The pre-analytical quality level of our laboratory from 2018 to 2020 was statistically analyzed in terms of four parts including sample collection, preservation and submission, centrifugation and quality inspection, which contained 17 indicators. 【Results】 Eleven sentinel indicators were established, and the order of peak value from high to low in three years was as follows: " label omission" rated at 0.000 62% (2020), " label error" 0.000 57% (2018), " inappropriate storage of samples before detection" 0.007 39 (2018), " unqualified application form for sample detection" 0.007 39 (2018). The causes were analyzed and relevant measures were taken. Six monitoring indicators were established, and the order of peak value from high to low in three years was as follows: " insufficient sample" rated at 0.002 59% (year 2018), " hemolysis" 0.002 80% (year 2020), " pale color of blood supernatant (diluted)" 0.000 86 (2018), " automatic sampling interfered by blood clot" 0.027 02% (2018). 【Conclusion】 The quality indexes of pre-analytical phase in our laboratory have reached the level of domestic and international clinical laboratories. The establishment of pre-analytical quality indicators and sentinel indicators, with effective analysis and application, can fully record and monitor the quality of each link before laboratory testing, which is helpful to timely identify risks, detect deviations, and quickly implement corrective and preventive measures, thus further ensure the safety of clinical blood use.