Primary cilium has attracted increasing attention in the biomedical field in recent years.It exists in the surface of various cells and is micro-sized,but its complex structures have not been clear.The primary cilium has an important role in sensory perception.Cells can receive extracellular mechanical and chemical signals through primary cilia,and primary cilia can assist in transferring signals into cells,followed by cellular responses.Recent studies have shown that the primary cilium also plays an important role in embryonic development and malignant transformation of cells.The investigation into primary cilia will facilitate the understanding of malignant transformation of cells and development of tumors.According to the related literature in recent years,this review summarizes the relationship between the primary cilium and common skin tumors,as well as potential targets for these tumors,so as to provide references and new sights for further researches.

Objective To evaluate the effect of mefformin on the human keratinocyte line HaCaT,and to explore its molecular mechanism.Methods HaCaT cells were divided into several groups to be treated with mefformin at different concentrations of 1,2,5,10,20,50 mmol/L for 24,48 and 72 hours.Cell counting kit-8 (CCK-8) assay was performed to evaluate the effect of metformin on the survival rate of HaCaT cells.After 48-hour treatment with metformin at concentrations of 0 (control group),0.5,1,2,5,10 mmol/L,flow cytometry was conducted to evaluate the effect of metformin on cell cycle and apoptosis.Western blot analysis was performed to determine the expression of cell proliferation-and differentiation-related proteins (keratin-16 [K16],K17,K1,involucrin),apoptosis-related proteins (Bax,Bcl-2) and AKT/mTOR/STAT3 pathway proteins.Enzyme-linked immunosorbent assay (ELISA) was conducted to detect the levels of interleukin (IL)-8,tumor necrosis factor (TNF)-α and IL-23 (inflammatory factors) in the culture supernatant of HaCaT cells.Statistical analysis was carried out with SPSS19.0 software using one-way analysis of variance for comparison of the above indices among the 0.5-,1-,2-,5-,10-mmol/L metformin groups and control group,repeated measures analysis of variance for comparisons among different time points or different metformin groups,least significant difference (LSD)-t test for multiple comparisons.Results CCK-8 assay showed that mefformin had inhibitory effects on the proliferation of HaCaT cells (F =116.87,P ＜ 0.05),and the cell survival rates gradually decreased along with the increase in the concentrations of mefformin.After 48-hour treatment with mefformin at concentrations of 0,0.5,1,2,5,10mmol/L,the proportion of HaCaT cells in G2/M phase gradually increased (5.55％ ± 1.03％,6.37％ ±0.93％,8.57％ ± 1.18％,10.05％ ± 0.60％,10.76％ ± 0.87％,13.63％ ± 1.41％,respectively,F =24.98,P ＜0.05),and the early apoptosis rate also gradually increased (0.78％ ± 0.71％,19.18％ ± 1.41％,25.67％ ±1.34％,28.45％ ± 0.92％,34.97％ ± 2.12％,40.41％ ± 1.49％,respectively,F =296.08,P ＜ 0.05).Along with the increase in the concentrations of metformin,there were increasing trends in the expression of K1 and the pro-apoptotic protein Bax (F =8.86,5.38 respectively,both P ＜ 0.05),while there were decreasing trends in the expression of K16,K17 and the apoptotic protein Bcl-2 (F =8.02,4.82,12.10 respectively,all P ＜ 0.05).There was no significant difference in the expression of involucrin among different metformin groups (F =0.57,P ＞ 0.05).After 48-hour treatment with mefformin at different concentrations,the levels of IL-8 and TNF-α in HaCaT cells significantly decreased (F =33.89,14.99 respectively,both P ＜0.05),while there was no significant change in the IL-23 level (F =2.12,P ＞ 0.05).Along with the increase in the concentrations of metformin,the expression of p-AKT,p-mTOR and p-STAT3 significantly decreased (F =11.38,0.35,4.38 respectively,all P ＜ 0.05),but there were no significant changes in the protein expression of AKT,mTOR and STAT3 (F =0.66,0.35,4.24 respectively,all P ＞ 0.05).Conclusion Metformin can inhibit the proliferation,promote the differentiation and apoptosis of HaCaT cells,and inhibit the secretion of inflammatory factors by regulating the AKT/mTOR/STAT3 signaling pathway.

Objective To explore the effect of Huoxue Xiaoyi Formula on ovarian function and pregnancy outcome in endometriosis rats. Methods One hundred female SD rats were fed adaptively for 7 days. According to body weight, 98 rats with a normal estrous cycle were randomly divided into the blank group (n=14), the sham operation group (n=14), the donor group (n=12), and the receptor group (n=58). A rat model of endometriosis was established using allogeneic endometrial transplantation. After the model was successfully established, according to the body weight, 56 rats in the receptor group were randomly divided into the model group, and the Huoxue Xiaoyi Formula low-, medium-, and high-dose groups, with 14 rats in each group. The Huoxue Xiaoyi Formula low-, medium-, and high-dose groups were given 5.86, 11.72, and 23.44 g/kg of Huoxue Xiaoyi Formula, and the blank group, the sham operation group, and the model group were given distilled water by gavage for 15 consecutive days. According to body weight, six rats were randomly selected to take abdominal aortic blood for determination of serum anti-Mullerian hormone (AMH) content by enzyme-linked immunosorbent assay. According to body weight, six rats were randomly selected in the remaining rats to undergo high-frequency small animal ultrasound examination to measure ovarian volume and ovarian artery blood supply (resistance index and pulsatility index), and follicle count were measured. The female mice were fed with male mice at a ratio of 2:1 to observe the pregnancy rate, live fetus number, still fetus number, and absorbed fetus number. The weights of uterus and fetus were measured, while the placental weight, fetal weight, body length, and tail length were measured for live fetus. Bone and visceral malformations were also detected for live fetus. Results There were no significant differences in serum AMH and ovarian volume among the groups. Compared with the model group, the numbers of follicles in the blank group, the sham operation group, and the Huoxue Xiaoyi Formula low-, medium-, and high-dose groups were increased; compared with the Huoxue Xiaoyi Formula low-dose group, the numbers of follicles in the blank group and the Huoxue Xiaoyi Formula medium-dose group were increased (P<0.05). Compared with the model group, the resistance index and pulsatility index of ovarian artery blood flow in the blank group, the sham operation group, and the Huoxue Xiaoyi Formula low-, medium-, and high-dose groups were decreased(P<0.05). There was no significant difference in pregnancy rate among the groups. Compared with the model group and the Huoxue Xiaoyi Formula low-dose group, the live fetus number and the weight of uterus and fetus in the blank group, the sham operation group, and the Huoxue Xiaoyi Formula medium- and high-dose groups were increased (P<0.05). There were no significant differences in the still fetus number, absorbed fetus number, placental weight, fetal weight, body length and tail length among the groups. No visceral or skeletal malformations were found in the live fetal rats among the groups.Conclusion In endometriosis model rats, the ovarian artery blood flow perfusion is insufficient, and the follicle count is reduced, affecting ovary function, thereby reducing the live fetus number. After applying the Huoxue Xiaoyi Formula, it can improve ovarian blood supply, increase the follicle count, and increase the live fetus number. The effect of increasing the live fetus number with the medium- and high-doses of Huoxue Xiaoyi Formula is more significant, and there is no reproductive toxicity.

Objective:This study is to investigate the parameter settings and imaging effects of Micro-CT scanning externalmalignancies during operation and summarize the characteristics of Micro-CT images regarding external soft tissue and tumors.Methods:According to the inclusion criteria, patients with externalmalignancies admitted to the Department of Plastic and Reconstructive Surgery of the First Medical Center of the PLA General Hospital from October 2018 to January 2020 were collected. Through the surgical resection of tumors, the isolated tumor specimens were divided into appropriate sizes (length × width × thickness not exceeding 5 cm×3 cm×3 cm), placed on the Micro-CT scanning bed, and scanned with different parameters. (1) Scanning with different times: with the fixedscanning voltage (50 kV for sarcoma, 70 kV for other tumors), the same specimen was scanned with 4min and 14min protocols respectively. (2) Scanning with different voltages: with the fixed scanning time (4 min), the same specimen was scanned with 30, 50, 70, and 90 kV voltages respectively. After comparing the imaging results with different scanning times and different scanning voltages, the optimal scanning parameters for different tumors were obtained. (3) The regions of interest for radiology were taken from the specimens, and pathological HE stained slides were prepared. Digital slices were obtained by precice automatic digital slice scanning system. The slices were compared with Micro-CT corresponding cross-sectional images and the radiological characteristics of different tissues were recorded.Results:A total of 27 patients were included/incorporated into the study and there were 15 males and 12 females, aged from 33 to 82, with an average age of 63.9 years old, including 16 cases of skin squamous cell carcinoma, 2 cases of basal cell carcinoma, and 2 cases of extramammary Paget′s disease, 1 case of melanoma, 3 cases of cutaneous fibrosarcoma, 1 case of angiosarcoma, and 2 cases of liposarcoma. A total of 65 groups were compared between Micro-CT images and pathology slides. (1) Scanning time: both 4 min and 14 min scanning times can obtain clear images which can meet requirements for morphology observation and identification of margins of tumors. (2) Scanning voltage: as for squamous cell carcinoma, the quality of images obtained by the scanning voltage of 30 and 50 kV is poor, while by the voltage of 70 and 90 kV is high. As for sarcoma, the quality of images obtained by the scanning voltage of 30 kV is poor; by the voltage of 50 kV is high and the contrast between tumor and subcutaneous fat is low by the scanning voltage of 70 and 90 kV. (3) The radiological morphology of external tumors and surrounding tissues are similar with the pathology slides. The boundary between skin and subcutaneous tissue is clear and the hair follicles are clearly distinguishable. The radiological characteristics of tumors are related to the type of tumors and the density of tumors, including high-density shadows, thicker than normal skin, no hair follicles and some small calcification points. The boundary between squamous cell carcinoma or sarcoma and normal skin or subcutaneous can be distinguished. As for the tumors in the skin, such as basal cell carcinoma, extra-mammary Paget′s disease, etc. the imaging results are poor.Conclusions:The Micro-CT scanning parameters need to be set according to the type of the tumor. The scanning time has little effect on the evaluation of the tumor. It is recommended to choose the protocol of 4 min during operation. The recommended scanning voltage of squamous cell carcinoma is 70 kV and the voltage of sarcoma is 50 kV. Micro-CT can clearly imaging the skin structure of soft tissue specimens on the body surface; configuration is identifiable for tumors that invade beyond skin and have a large heterogeneity of density compared with surrounding tissues; for tumors that do not exceed the dermis, the imaging effect is not good.

Objective:This study is to investigate the parameter settings and imaging effects of Micro-CT scanning externalmalignancies during operation and summarize the characteristics of Micro-CT images regarding external soft tissue and tumors.Methods:According to the inclusion criteria, patients with externalmalignancies admitted to the Department of Plastic and Reconstructive Surgery of the First Medical Center of the PLA General Hospital from October 2018 to January 2020 were collected. Through the surgical resection of tumors, the isolated tumor specimens were divided into appropriate sizes (length × width × thickness not exceeding 5 cm×3 cm×3 cm), placed on the Micro-CT scanning bed, and scanned with different parameters. (1) Scanning with different times: with the fixedscanning voltage (50 kV for sarcoma, 70 kV for other tumors), the same specimen was scanned with 4min and 14min protocols respectively. (2) Scanning with different voltages: with the fixed scanning time (4 min), the same specimen was scanned with 30, 50, 70, and 90 kV voltages respectively. After comparing the imaging results with different scanning times and different scanning voltages, the optimal scanning parameters for different tumors were obtained. (3) The regions of interest for radiology were taken from the specimens, and pathological HE stained slides were prepared. Digital slices were obtained by precice automatic digital slice scanning system. The slices were compared with Micro-CT corresponding cross-sectional images and the radiological characteristics of different tissues were recorded.Results:A total of 27 patients were included/incorporated into the study and there were 15 males and 12 females, aged from 33 to 82, with an average age of 63.9 years old, including 16 cases of skin squamous cell carcinoma, 2 cases of basal cell carcinoma, and 2 cases of extramammary Paget′s disease, 1 case of melanoma, 3 cases of cutaneous fibrosarcoma, 1 case of angiosarcoma, and 2 cases of liposarcoma. A total of 65 groups were compared between Micro-CT images and pathology slides. (1) Scanning time: both 4 min and 14 min scanning times can obtain clear images which can meet requirements for morphology observation and identification of margins of tumors. (2) Scanning voltage: as for squamous cell carcinoma, the quality of images obtained by the scanning voltage of 30 and 50 kV is poor, while by the voltage of 70 and 90 kV is high. As for sarcoma, the quality of images obtained by the scanning voltage of 30 kV is poor; by the voltage of 50 kV is high and the contrast between tumor and subcutaneous fat is low by the scanning voltage of 70 and 90 kV. (3) The radiological morphology of external tumors and surrounding tissues are similar with the pathology slides. The boundary between skin and subcutaneous tissue is clear and the hair follicles are clearly distinguishable. The radiological characteristics of tumors are related to the type of tumors and the density of tumors, including high-density shadows, thicker than normal skin, no hair follicles and some small calcification points. The boundary between squamous cell carcinoma or sarcoma and normal skin or subcutaneous can be distinguished. As for the tumors in the skin, such as basal cell carcinoma, extra-mammary Paget′s disease, etc. the imaging results are poor.Conclusions:The Micro-CT scanning parameters need to be set according to the type of the tumor. The scanning time has little effect on the evaluation of the tumor. It is recommended to choose the protocol of 4 min during operation. The recommended scanning voltage of squamous cell carcinoma is 70 kV and the voltage of sarcoma is 50 kV. Micro-CT can clearly imaging the skin structure of soft tissue specimens on the body surface; configuration is identifiable for tumors that invade beyond skin and have a large heterogeneity of density compared with surrounding tissues; for tumors that do not exceed the dermis, the imaging effect is not good.