OBJECTIVES: To study the effect of CDC20 knockdown on proliferation, migration and invasion of cervical cancer cells and its underlying mechanism. METHODS: CDC20 expression in cervical cancer tissues was analyzed using the TCGA database, and the protein expressions of CDC20 and β-Catenin in clinical specimens of cervical cancer and adjacent tissues were detected using immunohistochemistry. A dual target sgRNA2&7 sequence for CDC20 gene was designed for CDC20 gene knockdown in cervical cancer C33A cells using CRISPR/Cas9 technology, and CDC20 mRNA and protein expression levels in the transfected cells were detected using qRT-PCR and Western blotting. The changes in proliferation, cell cycle, apoptosis, migration and invasiveness of the transfected cells were evaluated using colony-forming assay, fluorescence activated cell sorting (FACS) and Transwell assay. In the animal experiment, naïve C33A cells and the cells with CDC20 knockdown were injected subcutaneously into the left and right axillae of nude mice (n=5) to observe tumor growth. The expressions of CDC20 and β-Catenin proteins in transfected cells and the xenograft were analyzed using Western blotting, and their interaction was confirmed by co-immunoprecipitation (CoIP) and immunofluorescence co-localization assays. RESULTS: Cervical cancer tissues expressed significantly higher CDC20 and β‑Catenin levels than the adjacent tissues. C33A cells with CDC20 knockdown showed reduced proliferation, increased apoptosis, and lowered migration and invasion abilities. CDC20 knockdown significantly suppressed the growth of C33A cell xenograft in nude mice, and the tumor-bearing mice did not exhibit obvious body mass changes. CDC20 and β-Catenin levels were both significantly lowered in C33A cells with CDC20 knockdown. Co-immunoprecipitation and co-localization assays confirmed the interaction between CDC20 and β‑Catenin. CONCLUSIONS CDC20 is highly expressed in cervical cancer tissues, and CDC20 knockdown can suppress proliferation, invasion, and metastasis while enhancing apoptosis of C33A cells, which is closely related with the regulation of the Wnt/β-Catenin signaling pathway.
Humans ; Uterine Cervical Neoplasms/metabolism* ; Female ; Cdc20 Proteins/genetics* ; Cell Proliferation ; Animals ; Cell Movement ; Neoplasm Invasiveness ; Apoptosis ; Mice, Nude ; beta Catenin/metabolism* ; CRISPR-Cas Systems ; Mice ; Cell Line, Tumor ; Gene Knockout Techniques ; Neoplasm Metastasis

[Purpose]To investigate the choice of medical institutions for the first-treatment of can-cer patients after diagnosis and the factors influencing it,so as to provide a reference basis for the rational allocation of health resources and the promotion of hierarchical diagnosis and treatment of cancer.[Methods]A total of 1 140 common cancer patients from 1 provincial,3 municipal and 6 county-level medical institutions in Shandong Province were included in the study,and a ques-tionnaire survey was conducted on their basic personal information and the selection of medical institutions.Chi-square test and Fisher exact test were used to compare different groups,and mul-ti-classification unordered Logistic regression were used to explore the factors influencing the se-lection of cancer patients.[Results]1 077 valid questionnaires were collected.The selection rates of first-treatment institutions for cancer patients were county hospitals(29.90％),city hospitals(35.28％),and provincial hospitals(34.82％),and the high level of medical care was the main reason for the selection of first-treatment institutions for cancer patients.The results of multi-clas-sification unordered Logistic regression showed that when choosing municipal and county medical institutions as the first treatment institution compared to choosing a county medical institution for the first treatment as a reference,compared to patients with stage Ⅰ tumors,patients with tumor stages at stage Ⅲ(OR=1.789,95％CI:1.014～3.158)and stage Ⅳ(OR=2.005,95％CI:1.179～3.409)were more likely to choose municipal medical institutions;and patients with annual house-hold income of 10 000～＜50 000 CNY(OR=0.625,95％CI:0.414～0.943)were less likely to choose mu-nicipal medical institutions compared to those with an annual household income of＜10 000 CNY.When choosing provincial and county medical institutions as the first treatment institution com-pared to patients in stage Ⅲ(OR=2.885,95％C1:1.549～5.372)and stage Ⅳ(OR=3.104,95％CI:1.724～5.586)compared to patients with stage Ⅰ tumors when choosing a county medical institu-tion for the first treatment was used as a reference;married(OR=2.248,95％CI:1.127～4.484)patients were more likely to choose provincial healthcare organizations than unmarried/divorced/widowed patients;patients with a family history of cancer compared to patients without a family history of cancer(OR=1.650,95％CI:1.115～2.441)were more inclined to choose provincial healthcare institutions;compared with patients＜45 years old,patients over 60 years old(OR=0.483,95％CI:0.242～0.962)were less inclined to choose provincial hospitals;and compared with patients who were not aware of knowledge related to cancer,those who were aware of knowledge related to cancer(OR=0.613,95％CI:0.441～0.852)patients were less inclined to choose provin-cial hospitals.[Conclusion]Vigorously carrying out publicity on cancer prevention and treatment knowledge,improving the cancer prevention and treatment literacy of cancer patients.strengthen-ing vertical cooperation among medical institutions at all levels,improving the provincial-munici-pal-county cancer prevention and treatment network,and promoting the sinking of high-quality medical resources are important ways to improve the cancer prevention and treatment capacity of counties and promote the work of hierarchical medical system.

[Purpose]To investigate the choice of medical institutions for the first-treatment of can-cer patients after diagnosis and the factors influencing it,so as to provide a reference basis for the rational allocation of health resources and the promotion of hierarchical diagnosis and treatment of cancer.[Methods]A total of 1 140 common cancer patients from 1 provincial,3 municipal and 6 county-level medical institutions in Shandong Province were included in the study,and a ques-tionnaire survey was conducted on their basic personal information and the selection of medical institutions.Chi-square test and Fisher exact test were used to compare different groups,and mul-ti-classification unordered Logistic regression were used to explore the factors influencing the se-lection of cancer patients.[Results]1 077 valid questionnaires were collected.The selection rates of first-treatment institutions for cancer patients were county hospitals(29.90％),city hospitals(35.28％),and provincial hospitals(34.82％),and the high level of medical care was the main reason for the selection of first-treatment institutions for cancer patients.The results of multi-clas-sification unordered Logistic regression showed that when choosing municipal and county medical institutions as the first treatment institution compared to choosing a county medical institution for the first treatment as a reference,compared to patients with stage Ⅰ tumors,patients with tumor stages at stage Ⅲ(OR=1.789,95％CI:1.014～3.158)and stage Ⅳ(OR=2.005,95％CI:1.179～3.409)were more likely to choose municipal medical institutions;and patients with annual house-hold income of 10 000～＜50 000 CNY(OR=0.625,95％CI:0.414～0.943)were less likely to choose mu-nicipal medical institutions compared to those with an annual household income of＜10 000 CNY.When choosing provincial and county medical institutions as the first treatment institution com-pared to patients in stage Ⅲ(OR=2.885,95％C1:1.549～5.372)and stage Ⅳ(OR=3.104,95％CI:1.724～5.586)compared to patients with stage Ⅰ tumors when choosing a county medical institu-tion for the first treatment was used as a reference;married(OR=2.248,95％CI:1.127～4.484)patients were more likely to choose provincial healthcare organizations than unmarried/divorced/widowed patients;patients with a family history of cancer compared to patients without a family history of cancer(OR=1.650,95％CI:1.115～2.441)were more inclined to choose provincial healthcare institutions;compared with patients＜45 years old,patients over 60 years old(OR=0.483,95％CI:0.242～0.962)were less inclined to choose provincial hospitals;and compared with patients who were not aware of knowledge related to cancer,those who were aware of knowledge related to cancer(OR=0.613,95％CI:0.441～0.852)patients were less inclined to choose provin-cial hospitals.[Conclusion]Vigorously carrying out publicity on cancer prevention and treatment knowledge,improving the cancer prevention and treatment literacy of cancer patients.strengthen-ing vertical cooperation among medical institutions at all levels,improving the provincial-munici-pal-county cancer prevention and treatment network,and promoting the sinking of high-quality medical resources are important ways to improve the cancer prevention and treatment capacity of counties and promote the work of hierarchical medical system.

Objective To organize the list of medicinal plants in Longzhong region;To conclude the species and distribution of medicinal plants in the region;To provide reference for the protection,development and utilization of TCM resources in the region.Methods The data of the fourth national census of TCM resources were obtained through the database of TCM resources census,and reference was made to the Chinese Botanical Records,Flora of the Loess Plateau,Gansu Herbal Resources Records,Gansu Provincial Standard of Chinese Materia Medica(2020 edition),Gansu Provincial Standard of Chinese and Tibetan Materia Medica(2020 edition),and other books and relevant literature supplementation,to summarize the medicinal plant species and distribution in Longzhong region.The status was summarized and analyzed.Results There are totally 178 families,829 genera and 2 101 species of medicinal plant resources in Longzhong region,mainly exist in angiosperms,gymnosperms and ferns and other groups,of which the dominant families are mainly concentrated in the Compositae,Rosaceae,Leguminosae,etc.The main medicinal parts for the whole grass class,mainly heat-clearing medicines,and 51 species of cultivated medicinal plants,including Astragalus membranaceus(Fisch.)Bge.var.mongholicus(Bge)Hsiao,Rosa rugosa Thunb.,Lonicera japonica Thunb.and so on.Conclusion Longzhong region is rich in plant resources and has many kinds of medicinal plants,which should be rationally developed and utilized on the basis of protection and vigorously develop characteristic TCM industry according to the geographical environment.

Background::Given that seizures may be triggered by vaccination, this study aimed to evaluate the risk and correlative factors of seizures in patients with epilepsy (PWE) after being vaccinated against coronavirus disease 2019 (COVID-19).Methods::This study retrospectively enrolled PWE who were vaccinated against COVID-19 in the epilepsy centers of 11 hospitals in China. We divided the PWE into two groups as follows: (1) patients who developed seizures within 14 days of vaccination were assigned to the SAV (with seizures after vaccination) group; (2) patients who were seizure-free within 14 days of vaccination were assigned to the SFAV (seizure-free after vaccination) group. To identify potential risk factors for seizure reccurence, the binary logistic regression analysis was performed. Besides, 67 PWE who had not been vaccinated were also included for elucidating the effects of vaccination on seizures recurrence, and binary logistic regression analysis was performed to determine whether vaccination would affect the recurrence rate of PWE who had drug reduction or withdrawal.Results::The study included a total of 407 patients; of which, 48 (11.8%) developed seizures within 14 days after vaccination (SAV group), whereas 359 (88.2%) remained seizure-free (SFAV group). The binary logistic regression analysis revealed that duration of seizure freedom ( P < 0.001) and withdrawal from anti-seizure medications (ASMs) or reduction in their dosage during the peri-vaccination period were significantly associated with the recurrence of seizures (odds ratio= 7.384, 95% confidence interval = 1.732–31.488, P = 0.007). In addition, 32 of 33 patients (97.0%) who were seizure-free for more than three months before vaccination and had a normal electroencephalogram before vaccination did not have any seizures within 14 days of vaccination. A total of 92 (22.6%) patients experienced non-epileptic adverse reactions after vaccination. Binary logistic regression analysis results showed that vaccine did not significantly affect the recurrence rate of PWE who had the behavior of ASMs dose reduction or withdrawal ( P = 0.143). Conclusions::PWE need protection from the COVID-19 vaccine. PWE who are seizure-free for >3 months before vaccination should be vaccinated. Whether the remaining PWE should be vaccinated depends on the local prevalence of COVID-19. Finally, PWE should avoid discontinuing ASMs or reducing their dosage during the peri-vaccination period.

ObjectiveTo clarify the spatial distribution characteristics of medicinal plant resources in Gansu province， analyze the causes， changing trends， and driving factors of the spatial differentiation， and thus lay a scientific basis for the rational development and sustainable development of medicinal plant resources in this province. MethodBased on the data of The Fourth National Survey of Chinese Medicine Resources， the richness and spatial distribution difference of medicinal plant resources in 87 counties （districts） of Gansu province were analyzed via the global spatial autocorrelation analysis， trend surface analysis， local spatial autocorrelation analysis， and hotspot analysis. Moreover， the correlation of vegetation type， soil texture， annual average temperature， annual average precipitation， and altitude with the spatial distribution pattern of the medicinal plant resources was discussed. ResultCounties （districts） with high or low richness of medicinal plant resources in Gansu province were respectively clustered together. To be specific， counties （districts） with high richness of the medicinal resources were mainly in southeastern Gansu， while those with low richness in northwestern Gansu. The leading driving factors affecting the cold and hot spots included vegetation type， soil texture， and average annual rainfall. ConclusionThe species richness of medicinal plant resources in Gansu province rises from west to east and from north to south. The natural driving factors are the key to the diversity and spatial distribution pattern of medicinal plant resources， which show significant influence on them.

Objective:To investigate the application of sentinel lymph node biopsy (SLNB) in early-staged cervical cancer by laparoscopy.Methods:It was a prospective, single-arm, single-center clinical study. Seventy-eight cases of cervical cancer patients were collected from July 2015 to December 2018 at the Fourth Hospital of Hebei Medical University. All the patients were injected with tracer into the disease-free block of cervical tissue after anesthesia by the same surgeon who learned sentinel lymph node (SLN) mapping technique in Memorial Sloan-Kettering Cancer Center, and underwent SLN mapping followed by complete pelvic lymphadenectomy. Moreover, all the dissected lymph nodes were stained with hematoxylin eosin staining (HE) pathological examination. Besides, the negative SLN on hematoxylin-eosin staining were detected by immunohistochemistry cytokeratin staining micro-metastasis. To analyze the distribution, detection rate, false negative rate the sensitivity and negative predictive value of the SLN in early-staged cervical cancer by laparoscopy, and explore the value of SLN mapping in predicting the lymph nodes metastasis in early-staged cervical cancer.Results:The overall detection rate of SLN in cervical cancer was 99% (77/78), bilateral detection rate was 87% (68/78). The average of 12.4 lymph node (LN) and 3.6 SLN were dissected for each patients each side. SLN of cervical cancer were mainly distributed in the obturator space (61.5%, 343/558), followed by external iliac (23.5%, 131/558), common iliac (7.3%, 41/558), para-uterine (3.8%, 21/558), internal iliac (2.2%, 12/558), para abdominal aorta (1.1%, 6/558), and anterior sacral lymphatic drainage area (0.7%, 4/558). Fourteen cases of LN metastasis were found among all 78 cases. There were a total of 38 positive LN, including 26 SLN metastasis and 12 none sentinel LN metastasis. Through immunohistochemical staining and pathological ultra-staging, 1 SLN was found to be isolated tumor cells (ITC), and 5 SLNs were found to be micro-metastases (MIC), accounting for 23% (6/26) of positive SLN. SLN mapping with pathological ultra-staging improved the prediction of LN metastasis in cervical cancer (2/14). Metastatic SLN mainly distributed in the obturator space (65%, 17/26), peri-uterine region (12%, 3/26), common iliac region (15%, 4/26), and external iliac region (8%, 2/26). The consistency of the diagnosis of lymph node metastasis by SLN biopsy and postoperative retroperitoneal lymph node metastasis showed that the Kappa value was 1.000 ( P<0.001), indicated that the metastasis status of SLN and retroperitoneal lymph node were completely consistent. The sensitivity, specificity, accuracy, false-negative rate, and negative predictive value of SLN biopsy in the diagnosis of lymph node metastasis were 100%, 100%, 100%, 0, and 100%, respectively. Conclusions:SLN in early-staged cervical cancer patients were mainly distributed in the obturator and external iliac space, pathalogical ultra-staging of SLN could improve the prediction of LN metastasis. Intraoperative SLN mapping is safe, feasible and could predict the state of retroperitoneal LN metastasis in early-staged cervical cancer. SLNB may replace systemic pelvic lymphadenectomy.

Objective To investigate the effect of SP600125 on the proliferation, cell cycle, apoptosis and invasion of human cervical cancer HeLa cells. Methods CCK-8 method was used to detect the proliferation of HeLa cells treated with different concentrations of SP600125 at different time points. The 20 μmol/L of SP600125 was determined for subsequent experiments. Cell proliferation ability was detected using plate clone formation assay; nuclear morphology was observed by DAPI staining; cell cycle and apoptosis were measured by flow cytometry; cell migration and invasion were detected by cell scratch and Transwell methods; the mRNA and protein levels of p53, Mad2L1 and CDC20 were measured by qRT-PCR and Western blot after SP600125 treatment at different time points. Results Compared with control group (0.1%DMSO), cells proliferative activity were reduced by 10, 20, 30, 40 and 50 μmol/L SP600125 treatment for 24h. Compared with control group, the rate of apoptosis was significantly increased in SP600125 treatment groups, and the cell proportion in G₂/M phase increased (P < 0.001), while the cell proportion in G₀/G₁ phases cells was reduced after SP600125 treatment for 24h and 48h (P < 0.001), and the clonal number, migration and invasion ability of HeLa cells also decreased significantly (P < 0.001). qRT-PCR and Western blot results showed a significant decrease in Mad2L1 mRNA and protein expression (P < 0.05) and a significant increase in p53 and CDC20 mRNA and protein expression (P < 0.01). Conclusion SP600125 can induce cell cycle arrest of cervical cancer HeLa cells in G₂/M phase by upregulating p53 and CDC20 and downregulating Mad2L1 expression, and promote cell apoptosis to inhibit cell proliferation, migration and invasion.

Objective:To investigate the correlation of monocyte/lymphocyte ratio (MLR) with the prognosis and adverse event in critically ill patients.Methods:Basic information of patients were extracted from Medical Information Mart for Intensive Care-Ⅲ (MIMIC-Ⅲ), including demographics, blood routine, biochemical indexes, systemic inflammatory response syndrome score (SIRS), sequential organ failure assessment (SOFA) score, and outcome, etc. MLR on the first day of intensive care unit (ICU) admission was calculated. The receiver operating characteristic curve (ROC curve) was applied to evaluate the prognostic value of MLR on the 30-day mortality and its cut-off value. According to the cut-off value, the patients were divided into two groups, and the differences between the groups were compared. Logistic regression model was used to analyze the relationship of MLR with 30-day mortality, continuous renal replacement therapy (CRRT), mechanical ventilation, the length of ICU stay, and total hospitalization time.Results:① A total of 43 174 critically ill patients were included. ROC curve showed that area under ROC curve (AUC) of MLR in predicting 30-day mortality was 0.655 [95% confidence interval (95% CI) was 0.632-0.687]. The cut-off value of MLR calculated according to the maximum Yoden index was 0.5. There were 16 948 patients with MLR ≥ 0.5 (high MLR group) and 26 226 patients with MLR < 0.5 (low MLR group). ② Compared with the low MLR group, the high MLR group had higher age, proportion of male, body mass index (BMI) [age (years old): 66.0 (51.7, 78.4) vs. 57.6 (27.1, 74.6), proportion of male: 57.2% vs. 52.5%, BMI (kg/m 2): 26.5 (22.5, 31.1) vs. 24.7 (14.3, 29.7)]. The high MLR group also had higher incidence of complications (hypertension: 49.2% vs. 44.6%, chronic heart failure: 32.6% vs. 21.7%, diabetes mellitus: 27.0% vs. 23.4%, chronic obstructive pulmonary disease: 21.5% vs. 16.1%, renal insufficiency: 19.3% vs. 13.1%), and higher white blood cell count (WBC), blood glucose, lactate (Lac), serum creatinine (SCr), SIRS score and SOFA score [WBC (×10 9/L): 13.8 (9.6, 19.2) vs. 11.5 (8.4, 15.6), blood glucose (mmol/L): 8.66 (6.88, 11.49) vs. 8.27 (6.55, 10.88), Lac (mmol/L): 2.2 (1.5, 3.7) vs. 2.1 (1.4, 3.3), SCr (μmol/L): 106.1 (70.7, 176.8) vs. 88.4 (70.7, 132.6), SIRS score: 3 (2, 4) vs. 2 (2, 3), SOFA score: 4 (2, 7) vs. 3 (1, 5)]. The 30-day mortality, and the proportion of patients with length of ICU stay > 5 days, total hospitalization time > 14 days, CRRT and mechanical ventilation > 5 days were significantly higher in high MLR group (30-day mortality: 20.0% vs. 8.3%, length of ICU stay > 5 days: 33.2% vs. 20.4%, total hospitalization time > 14 days: 33.7% vs. 16.2%, CRRT: 3.6% vs. 0.7%, mechanical ventilation > 5 days: 18.4% vs. 5.7%), with statistically significant differences (all P < 0.05). ③ After adjusted with the related factors, multivariate Logistic regression analysis showed that elevated MLR was an independent risk factor for increased 30-day mortality [odd ratio ( OR) = 1.54, 95% CI was 1.37-1.72, P < 0.001]. Moreover, the increased MLR was independently associated with the increased risk of usage of CRRT ( OR = 2.77, 95% CI was 2.18-3.51), mechanical ventilation > 5 days ( OR = 2.45, 95% CI was 2.21-2.72), the length of ICU stay > 5 days ( OR = 2.29, 95% CI was 2.10-2.49), and total hospitalization time > 14 days ( OR = 2.28, 95% CI was 2.08-2.49), all P < 0.001. Conclusions:Retrospective analysis of large sample shows that MLR elevation is an independent risk factor for 30-day mortality, usage of CRRT, prolonged mechanical ventilation time, prolonged hospitalization, prolonged length of ICU stay. MLR can be used for risk stratification of severe patients.

Nowadays, available phosphorus (P) deficiency in soil and weed resistance to herbicides have emerged as two severe limiting factors for sustainable agriculture. Therefore, it is of urgent needs to improve plant absorption/utilization ability of the soil P, seek phosphate (Pi)-alternative P fertilizers, and develop new forms of weed control systems. Phosphite (Phi), as a P resource of relatively high amount only less than Pi in Earth, can be converted to utilizable Pi uniquely in some bacterial species by oxidization via its specific dehydrogenase (PTDH), but inhibits plant growth and development. This implies that Phi might rather become a suitable P fertilizer for plants if introducing a PTDH detoxifier from bacteria. Herein, we created the transgenic tobaccos harboring a Pseudomonas PTDH gene (PsPtx) amplified from the soil metagenome previously. RT-PCR showed that the exotic PsPtx gene could express similarly in root, stem and leaf tissues of all transgenic lines. PsPtx transgenic tobaccos could utilize Phi by oxidization as the sole Pi supply, and also outperformed wild-type tobacco with a remarkably dominant growth under Phi stress conditions. Moreover, the PsPtx gene was preliminarily evaluated with a notable quality as a potential candidate of the selection marker in plant genetic transformation. Conclusively, PsPtx and its encoded phosphite dehydrogenase might be applicable for developing a dual system of plant phosphorus utilization and weed control using Phi as P fertilizer and herbicide, and provide an effectual solution to some obstacles in the current crop transgenic studies.
Oxidoreductases ; Phosphites ; Phosphorus ; Plants, Genetically Modified ; Weed Control