Background and purpose:In 2016 the National Cancer Institute(NCI)decided stopping to use NCI-60 cell lines for drug screening,suggesting that tumor cell lines were losing their value as a tool for drug discovery and basic research.The reason for NCI-60 cells'retirement'was that the preclinical studies based on traditional cellular and animal models did not obtain the corresponding expected efficacy in clinical trials.Since the major cancer behaviors,such as proliferation and metastasis,are fundamentally altered with long-term culture,the tumor cell lines are not representative of the characteristics of cancer in patients.Currently,scientists hope to create a new cancer model that are derived from fresh patient samples and tagged with details about their clinical past.Our purpose was to create patient-derived breast cancer primary cell lines as new cancer model for drug screening and basic research.Methods:Breast cancer tissues were collected in the Department of Breast Surgery,Affiliated Hospital of Guizhou Medical University.The collection of tumor tissue samples was approved by the Ethics Committee of the Affiliated Hospital of Guizhou Medical University(approval number:2022 ethics No.313),and the collection and use of tumor tissues complied with the Declaration of Helsinki.The primary breast cancer cell lines were isolated from the patient's breast cancer tissues and cultured in BCMI medium.After the cells proliferated,the media were replaced with DEME medium.Cell line STR genotyping was done to determine cell-specific genetic markers and identification.Clone formation assay and transplantation assay were done to analyze the ability of breast cancer primary cell lines to form tumors.Results:We created 6 primary breast cancer cell lines.The 6 primary breast cancer cell lines from the patients were tagged with the definitively clinicopathological features,clinical diagnosis,therapeutic regimens,clinical effectiveness and prognostic outcomes.The STR genotyping assays identified the genetic markers and determined the identities of the 6 primary breast cancer cell lines.Clone formation assays and transplantation assay showed that the proliferative capacities of the patient-derived primary breast cancer cell lines were significantly greater compared with the conventional breast cancer cell lines.Conclusion:We created a panel of 6 patient-derived primary breast cancer cell lines as new cancer model for drug screening and basic research in breast cancer.

Objective To observe the effects of"Sleep Recipe"on the behavior,brain tissue and central neurotransmitters of insomnia rats.Methods The male rats with SD were randomly divided into control group,model group,sleep formula group,and eszolam group,with 10 rats in each group,and the insomnia model was constructed by intraperitoneal injection of P-chlorphenylalanine(PCPA).After successful modeling,the control group and the model group were given saline gavage,and the medylom group and eszolam group were given drug gavage.The insomnia-like behavior of rats in each group was evaluated by pentobarbital sodium correction experiment and open field experiment,Hematoxlin and eosin(HE)staining observed the pathological changes of rat cerebral cortex,hippocampus,and hypothalamic tissue,and Enzyme-linked mmunosorbent assay(ELISA)was used to determine the expression levels of Gamma-aminobutyric acid(GABA),5-hydroxytryptamine(5-HT).Results The sleep latency of rats in the model group was significantly elongated(P<0.01),while the sleep time was less(P<0.01),and the mental state and fur color were poor,significantly decreased in body weight(P<0.01).Compared with the model group,the sleep latency was significantly shortened(P<0.01),the sleep duration was significantly prolonged(P<0.01),and the body mass was significantly increased(P<0.05,P<0.01);In the open field experiment,the total activity distance of rats in the model group increased,the average speed and central region residence time decreased(P<0.05),the total activity distance of rats in the Sleep Formula group and Eszolam group decreased significantly(P<0.05),the average speed increased and the central region residence time increased(P<0.05).HE results showed that the number of neurons,morphological structure and arrangement of neurons,such as cerebral cortex,hippocampus and hypothalamus in the model group,were damaged to varying degrees,and the sleep formula group and estazolam group were significantly improved.ELISA results showed that the expression of 5-HT and GABA in the cerebral cortex and hypothalamus of rats in the model group was significantly reduced(P<0.01,P<0.05),and the expression of GABA in hippocampal tissues was also significantly reduced(P<0.01).The protein expression levels of cerebral cortical GABA,hypothalamic GABA and 5-HT in the Sleep Formula group were significantly increased(P<0.01).Conclusion Sleep Formula can improve the mental state,restore normal body weight,improve sleep efficiency,and reduce anxiety and tension in insomnia rats.The mechanism may be related to increasing the content of 5-HT and GABA,and inhibiting the spread and conduction of hypothalamic and brainstem pro-awakening nuclei.

【Objective】 To investigate the effects of chlorogenic acid on the proliferation, migration and invasion of renal carcinoma A498 and 769-P cells and the possible molecular mechanism. 【Methods】 Human renal carcinoma A498 and 769-P cells were divided into control group and chlorogenic acid group (2 μL,1 μmol/L) and cultured for 72 h. The cell proliferation, invasion and migration were detected with MTT assay, Transwell assay and scratch test, respectively. The expressions of IL-1β, EPAS-1 and AKT/P65 signaling pathway related proteins were detected with ELISA, qRT-PCR and Western blot, respectively. 【Results】 Chlorogenic acid inhibited the proliferation, invasion and migration of renal carcinoma A498 and 769-P cells, and reduced the IL-1β level in the cell supernatant. Anti-IL-1β reduced the protein and mRNA expressions of EPAS-1, p-AKT and p-P65. Compared with the control group, the chlorogenic acid group had reduced mRNA and protein expressions of EPAS-1, p-AKT and p-P65 (P<0.05). 【Conclusion】 Chlorogenic acid can inhibit the invasion and metastasis of renal carcinoma cells, and its mechanism may be related to inhibiting the secretion of IL-1β, thereby inhibiting the AKT/P65/EPAS-1 pathway.

Osteoporotic vertebral compression fracture (OVCF) can lead to lower back pain and may be even accompanied by scoliosis, neurological dysfunction and other complications, which will affect the daily activities and life quality of patients. Vertebral augmentation is an effective treatment method for OVCF, but it cannot correct unbalance of bone metabolism or improve the osteoporotic status, causing complications like lower back pain, limited spinal activities and vertebral refracture. The post-operative systematic and standardized rehabilitation treatments can improve curative effect and therapeutic efficacy of anti-osteoporosis, reduce risk of vertebral refracture, increase patient compliance and improve quality of life. Since there still lack relevant clinical treatment guidelines for postoperative rehabilitation treatments following vertebral augmentation for OVCF, the current treatments are varied with uneven therapeutic effect. In order to standardize the postoperative rehabilitation treatment, the Spine Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized relevant experts to refer to relevant literature and develop the "Guideline for postoperative rehabilitation treatment following vertebral augmentation for osteoporotic vertebral compression fracture (2022 version)" based on the clinical guidelines published by the American Academy of Orthopedic Surgeons (AAOS) as well as on the principles of scientificity, practicality and advancement. The guideline provided evidence-based recommendations on 10 important issues related to postoperative rehabilitation treatments of OVCF.

ObjectiveTo understand the current situation of anxiety level and traditional Chinese medicine （TCM） nursing needs among residents during the COVID-19 epidemic， and to discuss the relationship between them，so as to provide references for maintaining their mental health. MethodsA total of 560 residents were recruited using convenience sampling method， and the selected individuals completed three questionnaires based on online platform， including self-designed general information questionnaire， Generalized Anxiety Disorder Scale （GAD-7）， and TCM nursing needs questionnaire. Then the correlation between anxiety level and TCM nursing needs was analyzed by Spearman correlation test. ResultsA total of 552 valid questionnaires were collected. The results of GAD-7 showed that the total score was （11.04±3.72）， and average score of each item was （1.58±0.53）. There were 328 cases （59.4%） without anxiety symptoms， 180 cases （32.6%） with mild anxiety， 33 cases （6.0%） with moderate anxiety， and 11 cases （2.0%） with severe anxiety. The analysis of TCM nursing needs questionnaire showed that the total score was （21.59±7.92）， and dominated by the needs of medical knowledge （2.90±1.15） and TCM dietary care （2.82±1.13）. The item score and total score of TCM nursing needs questionnaire of residents during the COVID-19 epidemic were positively correlated with GAD-7 total score （r=0.240~0.331， P<0.01）. ConclusionResidents suffer from different levels of anxiety during the COVID-19 epidemic， and TCM nursing needs is on the rise， indicating that the anxiety level of residents is correlated with the needs of TCM nursing.

Objective:To investigate the clinical value of the free fibula flap transplantation in repair of the defect of tibia and soft tissue with the help of augmented reality (AR) technology.Methods:From May, 2017 to May, 2019, 9 patients with tibial and soft tissue defects were treated. Before operation, CTA scan was performed on both shanks to obtain DICOM data of tibial bone defect. Images of the designed fibular flap and its blood supply model were imported into Sina software through computer virtual surgery assistant technology. With the AR technology, information of virtual fibula flap were projected onto the body surface at the donor site, and the operation was carried out under the precise positioning. In this group, the free fibular flap was harvested with an area of 6.0 cm×4.0 cm-12.0 cm×6.0 cm and the length of fibula was 6.0-13.5 cm. The free fibula flap were used to repair the tibial defect with 5.0-12.0 cm in length and soft tissue defect area at 5.0 cm×3.0 cm-10.0 cm×4.5 cm. Patients were followed-up to observe the survival of fibular flap and the functional recovery of the repaired lower limb, and evaluate the clinical effect.Results:All the fibular flaps survived without vascular crisis and without serious complications occurred at both of donor and recipient sites. After 8-12 months of follow-up, the flap was soft in texture and good in blood supply. The appearance of the repair site was not bloated. Callus began to form at the junction of fibula and tibia at 3-5 months and healed well in 8-12 months. No bone resorption, nonunion, loosening or falling off of screws were found. There was no pain in the shank of the recipient area. Patients could stand and walk freely, and the weight-bearing function was close to normal. According to Enneking system, the average score was 27 points; 7 cases were excellent and 2 cases were good.Conclusion:Application of AR technology in the repair of tibial bone defect with fibular flap transplantation has good clinical effect and has certain practical value.

Objective:To summarize and analyze the clinical data of Chinese patients with colony-stimulating factor 1 receptor (CSF1R)-related leukoencephalopathy, and clarify the phenotypic and genetic characteristics of Chinese patients.Methods:Medical history of patients with CSF1R-related leukoencephalopathy diagnosed from April 1, 2018 to January 31, 2021 in the department of neurology of 22 hospitals in China was collected, and scores of Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment Scale (MoCA), magnetic resonance severity scale were evaluated. Group comparison was performed between male and female patients.Results:A total of 62 patients were included, and the male-female ratio was 1∶1.95. The age of onset was (40.35±8.42) years. Cognitive impairment (82.3%, 51/62) and motor symptoms (77.4%,48/62) were the most common symptoms. The MMSE and MoCA scores were 18.79±7.16 and 13.96±7.23, respectively, and the scores of two scales in male patients (22.06±5.31 and 18.08±5.60) were significantly higher than those in females (15.53±7.41 , t=2.954, P=0.006; 10.15±6.26, t=3.328 , P=0.003). The most common radiographic feature was bilateral asymmetric white matter changes (100.0%), and the magnetic resonance imaging severity scale score was 27.42±11.40, while the white matter lesion score of females (22.94±8.39) was significantly higher than that of males (17.62±8.74 , t=-2.221, P<0.05). A total of 36 CSF1R gene mutations were found in this study, among which c.2381T>C/p.I794T was the hotspot mutation that carried by 17.9% (10/56) of the probands. Conclusions:The core phenotypic characteristics of CSF1R-related leukoencephalopathy in China are progressive motor and cognitive impairment, with bilateral asymmetrical white matter changes. In addition, there exist gender differences clinically, with severer cognitive impairment and imaging changes in female patients. Thirty-six CSF1R gene mutations were found in this study, and c.2381T>C/p. I794T was the hotspot mutation.

Objective:To investigate the clinical value of augmented reality (AR) technology combined with Doppler ultrasound in repairing soft tissue defects around ankle with posterior tibial artery perforator flap.Methods:From June, 2017 to June, 2020, 13 patients with soft tissue defects around ankle were repaired with the posterior tibial artery perforator flap sized 3.0 cm×7.0 cm-5.0 cm×10.0 cm. Preoperative CTA scan was performed to obtain the 3-dimensional image of the blood supply of the donor site. The origin and direction of blood supply, length of pedicle, diameter and the location of perforator were determined. An image fusion software based on Android mobile phone was used from the body surface to realize the navigation by AR, and to mark the locations of the points where blood vessels ran and passed by. At the same time, Doppler ultrasound was employed to confirm the navigation. The optimal puncture fulcrum was used as the flap rotation point in design of the flap. After the flap had been designed, the flap was harvested and the defect was repaired. Outpatient clinic follow-up was performed to evaluate the survival of flap and the function of ankle.Results:All 13 flaps survied without vascular issues. After 6-16 (average 10) months follow-up, all flaps survived. The flaps appeared soft with good blood supply. The colour of the repaired sites were basically the same as normal skin. According to American Orthopedic Foot and Ankle Society (AOFAS) , the ankle function was excellent in 9 patients and good in 4 patients. There was no obvious scar formation on the donor sites.Conclusion:The application of AR technology combined with Doppler ultrasound in the surgical planning of posterior tibial artery perforator flap has advantages in easy to operate in clinical practice. It can reduce the risk of surgery, and has significance in clinical practice.

Anti-Sry-like high mobility group box (SOX) 1 antibodies (abs) are partly characterized onconeural autoantibodies (autoabs) due to their correlation with neoplastic diseases. AntiSOX1 abs are associated with various clinical manifestations, including Lambert-Eaton myasthenic syndrome (LEMS) and paraneoplastic cerebellar degeneration (PCD). However, the clinical characteristics of patients with anti-SOX1 abs have not been described in detail. This review systematically explores the reported patients with anti-SOX1 abs and analyzes these cases for demographic characteristics, clinical features, coexisting neuronal autoabs, neuroimaging findings, treatment, and clinical outcomes. In addition, considering that PCD is the most common paraneoplastic neurological syndrome and that the association between PCD and anti-SOX1 abs remains unclear, we focus on the presence of autoabs in relation to PCD and associated tumors. PCD-associated autoabs include various intracellular autoabs (e.g., anti-Hu, anti-Yo, anti-Ri, and anti-SOX1) and cell-surface autoabs (anti-P/Q-type voltage-gated calcium channel). Commonly involved tumors in PCD are small-cell lung cancer (SCLC), gynecological, and breast tumors. LEMS is the most common clinical symptom in patients with antiSOX1 abs, followed by PCD, and multiple neuronal autoabs coexist in 47.1% of these patients.SCLC is still the predominant tumor in patients with anti-SOX1 abs, while non-SCLC is uncommon. No consistent imaging feature is found in patients with anti-SOX1 abs, and there is no consensus on either the therapy choice or therapeutic efficacy. In conclusion, the presence of anti-SOX1 abs alone is a potential predictor of an uncommon paraneoplastic neurological disorder, usually occurring in the setting of LEMS, PCD, and SCLC. The detection of anti-SOX1 abs contributes to an early diagnosis of underlying tumors, given the diversity of clinical symptoms and the absence of characteristic neuroimaging features.

Objective:To investigate and analyze the current situation, screening, clinical characteristics, prevention and treatment of bleeding esophageal varices in cirrhotic patients with portal hypertension in Tibet region.Methods:Clinical data of cirrhotic patients with portal hypertension through March 2017 to February 2020 from Tibet region were collected and analyzed retrospectively.Results:511 cases with liver cirrhosis were included in the study, of which 185 cases (36.20%) had compensated cirrhosis and 326 cases (63.80%) had decompensated cirrhosis. Further analysis of the etiological data of liver cirrhosis showed that 306 cases (59.88%) were of chronic hepatitis B, 113 cases (22.11%) of alcoholic liver disease, and 68 cases (13.31%) of chronic hepatitis B combined with alcoholic liver disease. Among patients with compensated liver cirrhosis, 48 cases (25.95%) underwent endoscopic examination of which 33 diagnosed as high-risk variceal bleeding. However, none of these 33 cases had received non-selective β-blocker therapy, and only four patients had received endoscopic variceal banding therapy. Among patients with decompensated liver cirrhosis, 83 cases (25.46%) had a history of upper gastrointestinal bleeding, 297 cases (91.10%) had ascites, 23 cases (7.05%) had hepatic encephalopathy, and 3 cases (0.92%) had hepatorenal syndrome. Among the patients with a history of upper gastrointestinal bleeding, 42 cases (50.60%) had received secondary preventive treatment for bleeding esophageal varices, including 39 cases of endoscopic treatment, 1 case of endoscopic combined drug treatment, 3 cases of interventional treatment, and 2 cases of surgical treatment.Conclusion:Chronic hepatitis B and alcoholic liver diseases are the main causes of liver cirrhosis in Tibet region. Moreover, this region lacks screening, prevention and treatment for bleeding esophageal varices in cirrhotic patients with portal hypertension. Therefore, it is necessary to increase the screening of high-risk groups to prevent and improve the first-time bleeding, and promote multidisciplinary team to prevent and treat re-bleeding.