AIM:To investigate the relationship between the expression of phosphatidylinositol 3-kinase cata-lytic subunit alpha(PIK3CA)gene in circulating tumor cells(CTCs)and the prognosis of colorectal cancer patients,and to construct a corresponding predictive model.METHODS:This retrospective study included 110 newly diagnosed and treated colorectal cancer patients at Guangzhou Red Cross Hospital from January 2019 to December 2021.Peripheral blood samples were collected and tested for CTC and PIK3CA gene at first visit.We determined the expression levels of PIK3CA gene in CTCs of different subtypes by typing and counting CTCs.Patients were randomly divided into training group(n=77)and validation group(n=33)in a ratio of 7∶3.Univariate and multivariate Cox regression analyses were per-formed to assess the associations between clinicopathological factors and patient prognosis.A nomogram model was con-structed to predict survival of patients at 6,12 and 24 months.The model performance was evaluated and validated using the concordance index(C-index),receiver operating characteristic(ROC)curve,and decision curve analysis(DCA)in the validation group.And then,the clinical application value of the model was further evaluated using the DCA curve.RESULTS:Univariate analysis showed that N1 and N2 stage,M stage(M1),carcinoembryonic antigen(CEA)level,liver metastasis,and lung metastasis were independent risk factors for prognosis(P<0.01).Multivariate analysis showed that age,gender,N1 stage,TNM stage(IIC,IIIA,IV),CEA level,liver metastasis,lung metastasis,and PIK3CA ex-pression in mixed CTCs were correlated with prognosis(P<0.05).The nomogram model based on PIK3CA expression demonstrated excellent predictive performance in the validation group,with areas under the ROC curve(AUC)of 0.952(6 months),0.931(12 months),and 0.957(24 months).The DCA curve showed that the model had a high net benefit across a wide range of decision thresholds.CONCLUSION:The different expression levels of PIK3CA gene in CTC are correlated with the survival rate of patients.Especially in mixed CTCs,high expression of PIK3CA gene is associated with shorter overall survival.The clinical prediction model nomogram has good clinical application value in predicting the sur-vival rate of colorectal cancer patients.The decision curve analysis also shows that the model has good net benefits and clinical application value.

AIM:To investigate the relationship between the expression of phosphatidylinositol 3-kinase cata-lytic subunit alpha(PIK3CA)gene in circulating tumor cells(CTCs)and the prognosis of colorectal cancer patients,and to construct a corresponding predictive model.METHODS:This retrospective study included 110 newly diagnosed and treated colorectal cancer patients at Guangzhou Red Cross Hospital from January 2019 to December 2021.Peripheral blood samples were collected and tested for CTC and PIK3CA gene at first visit.We determined the expression levels of PIK3CA gene in CTCs of different subtypes by typing and counting CTCs.Patients were randomly divided into training group(n=77)and validation group(n=33)in a ratio of 7∶3.Univariate and multivariate Cox regression analyses were per-formed to assess the associations between clinicopathological factors and patient prognosis.A nomogram model was con-structed to predict survival of patients at 6,12 and 24 months.The model performance was evaluated and validated using the concordance index(C-index),receiver operating characteristic(ROC)curve,and decision curve analysis(DCA)in the validation group.And then,the clinical application value of the model was further evaluated using the DCA curve.RESULTS:Univariate analysis showed that N1 and N2 stage,M stage(M1),carcinoembryonic antigen(CEA)level,liver metastasis,and lung metastasis were independent risk factors for prognosis(P<0.01).Multivariate analysis showed that age,gender,N1 stage,TNM stage(IIC,IIIA,IV),CEA level,liver metastasis,lung metastasis,and PIK3CA ex-pression in mixed CTCs were correlated with prognosis(P<0.05).The nomogram model based on PIK3CA expression demonstrated excellent predictive performance in the validation group,with areas under the ROC curve(AUC)of 0.952(6 months),0.931(12 months),and 0.957(24 months).The DCA curve showed that the model had a high net benefit across a wide range of decision thresholds.CONCLUSION:The different expression levels of PIK3CA gene in CTC are correlated with the survival rate of patients.Especially in mixed CTCs,high expression of PIK3CA gene is associated with shorter overall survival.The clinical prediction model nomogram has good clinical application value in predicting the sur-vival rate of colorectal cancer patients.The decision curve analysis also shows that the model has good net benefits and clinical application value.

Objective To analyze the characteristics of imported malaria epidemic from overseas in Wuhan, to explore the management mechanism of on-site cases, and to accumulate experience for the treatment of imported malaria in large cities after malaria elimination. Methods The epidemiological data on imported malaria from abroad during the period of malaria elimination (2010-2019) in Wuhan were collected. The gender, age and severe illness-related factors of the cases were analyzed. Based on the characteristics of the epidemic and the current situation of prevention and control, the content and experience of the “Municipal-District 24-7” case mechanism were discussed. Results The medical resources in Wuhan were the best in the central region, resulting in a large number of imported malaria cases, with a total of 474 cases reported from 2010 to 2019 (40.79% of the total number of cases in Hubei Province), including 359 cases of falciparum malaria, 36 severe cases and one death (the death rate was 0.28%). The patients were mainly young and middle-aged men aged 20 to 49 years old (97.26%). There were many referral cases (40.30%), and there was no seasonal clustering of cases reported. The undiagnosed proportion at the first visit was 44.85%, and the time of attack-diagnosis was 4 days or more in 61.00% of cases. The occurrence of severe cases was related to unconfirmed diagnosis at the first visit (χ²=35.46, P<0.001) and attack-diagnosis time (Z=-6.49, P<0.001). Conclusion Imported malaria occurs frequently in Wuhan, mainly falciparum malaria. However, “Municipal-District 24-7” case mechanism has effectively curbed the occurrence of severe and death cases and provided valuable experience for case management in similar cities in China.

Qingweisan is one of the classical prescriptions commonly used in the treatment of oral diseases. By means of Bibliometrics， the authors collected the ancient books on Qingweisan and sifted out 411 valid data， involving 116 classics of traditional Chinese medicine. The historical origin， drug composition， indications， principle of composition， dosage，and preparation of Qingweisan were statistically analyzed， and it was found that the prescription originated from the Treatise on Spleen and Stomach（《脾胃论》） by LI Dongyuan and is composed of Rehmanniae Radix， Angelica Sinensis， Cortex Moutan， Coptidis Rhizoma and Cimicifugae Rhizoma， with the functions of clearing stomach， purging fire， cooling blood and dispersing depression. And Qingweisan was mainly used to treat toothache， headache， and preference for cold and aversion to heat caused by "excessive heat in yang brightness meridian". There were 352 indications recorded， most of which followed LI Dongyuan's theory and the expanded indications reached 70 kinds. Specifically， toothache （132） was the most， accounting for 22.49% of the total indications， followed by headache （60， 10.22%）. In addition， Qingweisan was widely used in modern clinical practice for multiple system diseases， among which oral system （197） was dominant， accounting for 72.69%， followed by skin system （28， 10.33%） and digestive system （27， 9.96%）. Although the indications were wide， the pathogenesis always belonged to "upward attack of stomach fire". Through the excavation and statistical analysis of the ancient books on Qingweisan and its modern clinical application， the authors aimed to provide a more scientific reference for the research and application of classical famous prescriptions.

Objective: To evaluate the role of extracellular signal-regulated kinase (ERK) signaling pathway in intrathecal dexmedetomidine-induced reduction of spinal cord ischemia-reperfusion (I/R) injury in rats. Methods: Eighty clean-grade male Sprague-Dawley rats, aged 9-10 weeks, weighing 300-350 g, were divided into 4 groups (n=20 each) using a random number table method: sham operation group (group S), spinal cord I/R group (group I/R), dexmedetomidine group (group D), and dexmedetomidine plus ERK signaling pathway blocker PD98059 group (group P). Spinal cord ischemia was produced by cross-clamping of the abdominal aorta distal to the left renal artery for 25 min followed by reperfusion to establish the model of spinal cord I/R injury.Dexmedetomidine 1 μg/kg was intrathecally injected at 20 min before establishing the model in D and P groups, PD98059 2 mg/kg was given via the tail vein at the same time in group P, and the equal volume of normal saline was given instead in S and I/R groups.Five rats were selected at 6, 8, 10 and 12 h of reperfusion, and the modified Basso, Beattie, Bresnahan (BBB) scale was used to assess the hindlimb locomotor function.Five rats were sacrificed after assessing the locomotor function at 6 h of reperfusion, and the L_3-5 segments of the spinal cord were taken for determination of cell apoptosis (by TUNEL) and expression of phosphorylated ERK (p-ERK) (by Western blot). The apoptosis index was calculated. Results: Compared with group S, the BBB scores were significantly decreased at each time point of reperfusion, the apoptosis index was increased, and the expression of p-ERK was up-regulated in the other three groups (P<0.05). Compared with I/R group, the BBB scores were significantly increased at each time point of reperfusion, the apoptosis index was increased, and the expression of p-ERK was up-regulated in group D, and no significant change was found in the apoptosis index or p-ERK expression in group P (P>0.05). Compared with group D, the BBB scores were significantly decreased at each time point of reperfusion, the apoptosis index was increased, and the expression of p-ERK was down-regulated in group P (P<0.05). Conclusion The mechanism by which intrathecal dexmedetomidine reduces spinal cord I/R injury is related to activating ERK signaling pathway in rats.

Objective To study the expression and the function of DKK2 and to explore its potential mechanisms in breast cancer.Methods The expression of DKK2 was detected by RT-PCR in normal breast tissues and breast cancer cells.we have transfected DKK2 into breast cancer cell lines MDA-MB-231 and MCF-7.The cells before transfection were used as control group and marked with Vector.The cells after transfection were used as experimental group and marked with DKK2.Furthermore by qRT-PCR and Western-blot,the expression of DKK2,Notch signaling pathway and related factors were analyzed.We also detected the function of DKK2 by cloning assay,Transwell assay and proliferation assay.Results No expression of DKK2 was found in breast cancer cell lines MDA-MB-231 or MCF-7,with relatively high expression in normal breast tissue.The number of apoptotic cells was 2.57±1.18 before transfectionin in cell line MDA-MB-231,and 49.53±8.27 after transfection.The difference was statistically significant between the two groups (P＜0.005).The relative colony formation rate of MDA-MB-231 cells and MCF7 cells after transfection accounted for 20.44％ and 15.21％,respectively.The difference was statistically significant by t test.The number of apoptosis cells in MB231/DKK2 group was 49.53± 8.27 and that in MB231 / Vector group was 2.57±1.18.The difference was statistically significant (P＜0.005).The number of migrated cells in MB231/DKK2 group was 112.0±8.1 and that in MB231/Vector group was 178.0±12.0.The difference was statistically significant (P＜0.005).The mRNA expression of Notch 1 in group MB231/Vector was recorded as 1.The mRNA expression of JAG1 in MB231/DKK2 group was 0.2891.The difference was also statistically significant (P＜0.005).Conclusions Restored expression of DKK2 in silenced breast cells suppresses breast cancer cell proliferation and migration through repressing Notch signaling.DKK2-Notch signaling pathway may be its potential molecular mechanism to function in breast cancer.DKK2 may be one of the target genes for early diagnosis and treatment of breast cancer.