Objective:To explore the effect and mechanism of 5-Aminolevulinic Acid-Photodynamic Therapy (5-ALA-PDT) on the apoptosis of the human colonic carcinoma HT-29 cells.Methods:HT-29 cells were cultured in vivio and divided into four groups: blank control group, 5-ALA group, PDT group and 5-ALA-PDT group.The control group was not given photosensitizer and light treatment; 5-ALA group was given photosensitizer ; PDT group was given light treatment; 5-ALA-PDT group was given photosensitizer and light treatment at the same time. Flow cytometry was used to observe the apoptosis of HT-29 cells. Reverse transcription polymerase chain reaction (RT-PCR) was used to observe the expression of B-type lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) in HT-29 cells. Ultraviolet spectrophotometry was used to detect the expression of Caspase-3. Results:The apoptotic rate of 5-ALA-PDT group was significantly higher than that of blank control group, 5-ALA group and PDT group ( P<0.05). Compared with the blank control group, 5-ALA-PDT group and PDT group, the expression of Bcl-2 in the 5-ALA-PDT group was statistically significant ( P<0.05), but there was no significant difference in Bax expression among the four groups ( P>0.05). The expression of Bax/Bcl-2 in 5-ALA-PDT group was significantly higher than that in blank control group, 5-ALA group and PDT group ( P<0.05). The expression of Caspase-3 in 5-ALA-PDT group was significantly higher than that in blank control group, 5-ALA group and PDT group ( P<0.05). Conclusions:5-ALA-PDT can induce apoptosis of HT-29 cells, and its mechanism may be related to the induction of apoptosis through Bax/Bcl-2 pathway.

Objective:To investigate the accumulation of porphyrin metabolites [uroporphyrinogen (UP) Ⅰ and coproporphyrinogen (CP) Ⅲ] induced by 5-aminolevulinic acid (5-ALA) in the urine of rats with colorectal cancer.Methods:The rat model of colorectal cancer was established by dimethylhydrazine (DMH). Urine samples were collected from 30 colorectal cancer rats (colorectal cancer group) and 30 normal rats (normal group). Each animal was given 5-ALA (50 mg/kg) by gavage, and urine was collected after 2, 4, 6 and 8 h. The contents of urinary porphyromogen Ⅰ and porphyromogen faecalis Ⅲ in urine were detected by high performance liquid chromatography (HPLC).Results:There was no significant difference in the contents of UP Ⅰ and CP Ⅲ in urine between colorectal cancer group and normal group before oral administration of 5-ALA ( P>0.05). After oral administration of 5-ALA, the contents of UP Ⅰ and CP Ⅲ in urine of colorectal cancer group were significantly higher than those of normal group ( P<0.05). The contents of UP Ⅰ and CP Ⅲ in urine of colorectal cancer group reached the highest value at 4 hours. According to the receiver operating characteristic (ROC) curve drawn from 4-hour test results, the threshold value of UP Ⅰ for colorectal cancer diagnosis was 50.43 μmol/g, with corresponding sensitivity 96.7%, and the specificity 63.3%, respectively. The threshold value of CP Ⅲ for colorectal cancer diagnosis was 108.85 μmol/g, with corresponding sensitivity 66.7%, and the specificity 86.7%, respectively. Conclusions:The accumulation of porphyrin metabolites induced by 5-ALA in the urine of rats with colorectal cancer is significant. The porphyrin metabolites in urine may be a new tumor marker of colorectal cancer, which provides an experimental basis for the diagnosis of colorectal cancer.

Objective:To investigate the changes of porphyrin metabolites in urine of patients with colorectal cancer before and after operation and their correlation with prognosis.Methods:One hundred patients with colorectal cancer were collected in First Affiliated Hospital of Hebei North University from June 2016 to December 2016, urine was collected before operation, 1 week after operation, 1 year after operation and before recurrence. The contents of urinary porphyrin metabolites of uroporphyrinogenI (UP Ⅰ) and coproporphyrinogen Ⅲ(CP Ⅲ) were detected by high performance liquid chromatography. Toanalyse the changes of UPⅠ and CPⅢ levels before and after operaction of colorectal cancer and their correlation with clinicopathologicalcharacteristics,and the recurrence and metastasis after operation.Results:The levels of UPⅠ and CPⅢ in urine of patients with colorectal cancer after operation were significantly lower than those before operation [(66.80 ± 17.62) μmol/g vs. (35.58 ± 9.32) μmol/g, (20.14 ± 3.14) μmol/g vs. (10.38 ± 0.85) μmol/g] ( P<0.05). The levels of UP Ⅰ and CP Ⅲ in urine of patients with Dukes C/D stage were significantly higher than those with Dukes A/B stage [(45.26 ± 5.26) μmol/g vs. (28.56 ± 3.45) μmol/g, (86.57 ± 6.58) μmol/g vs. (52.48 ± 3.36) μmol/g], the levels of UP Ⅰand CPⅢ in urine of patients with lymph node metastasis were significantly higher than those without lymph node metastasis [(45.44 ± 5.46) μmol/g vs. (30.27 ± 6.07) μmol/g, (86.67 ± 6.87) μmol/g vs. (56.10 ± 11.08) μmol/g], there were significant differences ( P<0.05). Urinary levels of UPⅠ and CPⅢ were independent risk factors for recurrence and metastasis of colorectal cancer after operation ( OR=1.149 and 1.065, P<0.05). Conclusions:Porphyrin metabolites (UPⅠ and CPⅢ) in urine may serve as a new marker for assessing colorectal cancer.

Objective:To analyze the cardio-pulmonary ultrasound features of cardiogenic pulmonary edema (CPE) and pneumonia in adults with acute dyspnea, and to construct a differential diagnosis model.Methods:Seven hundred and forty-three patients with sudden acute dyspnea admitted to Hebei General Hospital from November 2018 to May 2022 were retropectively included. Ultrasonographer A performed lung ultrasound with 12 zone method, and interpreted and recorded the ultrasonic signs (including A-lines area, B-lines area, consolidation area and pleural effusion area) together with ultrasonographer B. According to the ultrasonic characteristics of the whole lung, it was divided into A-profile and B-profile. According to the continuity and symmetry of the distribution of B-lines in bilateral lung fields, it could be divided into bilateral lung continuous and discontinuous B-profile, bilateral lung symmetric and asymmetric B-profile. Left ventricular ejection fraction (LVEF), left ventricular filling pressure (E/e′), right ventricular dilatation, tricuspid annular systolic displacement (TAPSE) and inferior vena cava diameter (IVCD) were evaluated by echocardiography, and all the indexes were transformed into binary variables. According to the final clinical diagnosis and treatment results, the disease was divided into CPE group and pneumonia group. Binary Logistic regression model was used to screen independent influencing factors, and partial regression coefficient β value was used as a weight to assign a score, and a differential diagnosis model was established based on the total score. The predictive value of the model was evaluated by the receiver operating characteristic curve (ROC) and area under curve (AUC). After the model was built, 30 patients with CPE or pneumonia were independently collected by ultrasonographer C as external validation data, which were included in the model to draw ROC curve and evaluate the differential diagnosis efficiency of the model. The consistencies between ultrasonographer A and B, A and C in observing lung ultrasound were explored.Results:A total of 743 patients from 43 clinical departments were included, including 246 cases in CPE group and 497 cases in pneumonia group. Multivariate logistic regression analysis showed that bilateral lung continuous B-profile, bilateral lung symmetric B-profile, ≥1 pleural effusion area, LVEF<50%, E/e′>14 were the risk factors for CPE (all OR>1, P<0.05), and ≥1 consolidation area and ≥1 pleural sliding disappearance area were the protective factors for CPE (all OR>1, P<0.05). The sensitivity, specificity and AUC of combined cardio-pulmonary ultrasound index β value weight score in the differential diagnosis of CPE and pneumonia were 0.939, 0.956 and 0.986, respectively. The AUC of external validation data was 0.904. Ultrasonographer A and B, A and C had good consistency in the interpretation of lung ultrasound signs ( P<0.05). Conclusions:The differential diagnosis model based on combined cardio-pulmonary ultrasound indexes has high differential diagnosis efficiency for CPE and pneumonia, and can be used in bedside cardio-pulmonary ultrasound practice.

Objective To compare the accuracy of bedside lung ultrasound in emergency(BLUE)and combined cardiac-lung and additional ultrasound(CLAUS)for diagnosing the causes of acute dyspnea.Methods Totally 1 016 patients with acute dyspnea were retrospectively enrolled and divided into cardiogenic pulmonary edema group(n=268),pneumonia group(n=574),pneumothorax group(n=33),pulmonary embolism group(n=67)and CAD(chronic obstructive pulmonary disease/asthma/diaphragmatic dysfunction)group(n=74)according to the causes of acute dyspnea.The findings of CLAUS protocol were compared among groups,and the accuracy of BLUE and CLAUS protocol for diagnosing the causes of acute dyspnea were also compared.Results CLAUS showed that B-B and B-C were the most common modes in cardiogenic pulmonary edema group,while A-B/A-C/B-A/B-B/B-C/C-C modes were common in pneumonia group,and A-A mode was the most common in pneumothorax group,pulmonary embolism group and CAD group.Significant differences of the manifestations of pulmonary ultrasound,pleural feature of anterior chest wall,left/right cardiac insufficiency and abnormal inferior vena cava diameter were found among groups(all P＜0.05).The accuracy of BLUE and CLAUS protocol for diagnosing the causes of acute dyspnea was 86.91%(883/1 016)and 94.49%(960/1 016),respectively,the latter was higher than the former(χ2=34.587,P＜0.05).Conclusion CLAUS protocol could be used to effectively diagnose the causes of acute dyspnea,with higher accuracy than BLUE protocol.

Objective To standardize the management of clinical trials in our hospital,reduce the incidence of protocol deviations,and provide a reference for improving the quality of clinical trials.Methods The healthcare failure mode and effect analysis(HFMEA)method was used to determine the potential failure modes of the current protocol deviation.The frequency,severity and detectability of failure modes were quantified and evaluated.The risk priority number(RPN)was calculated and the corresponding improvement measures were proposed.The RPN values before and after the implementation of HFMEA were statistically analyzed to evaluate the improvement effect.Results After the implementation of HFMEA activities,the RPN values of 14 potential failure modes decreased significantly(P<0.05);The risk level of 12 potential failure modes decreased.The HFMEA team members'ability in finding and solving problems,communication and cooperation were significantly improved.Conclusions The implementation of HFMEA activities contributes to the management of protocol deviation in clinical trials,can effectively reduce the occurrence of protocol deviation,and provides experience for improving the quality of drug clinical trials.

OBJECTIVE To characterize the chemical constituents of the cladodes and fruits of Opuntia Milpa Alta by using UPLC-QE-Orbitrap MS, and investigate the antioxidant activity, and explore the relationship between the constituents and biological activity. METHODS The UPLC HSS T3 C18 column(2.1 mm×10 mm, 1.8 μm) was used. Acetonitrile and water with 0.1% formic acid was used for gradient elution at a 0.2 mL·min–1 flow rateas. Heated electrospray ion source was used for primary and secondary mass spectrometry data acquisition in positive and negative ion modes, respectively. Database and literature reports were used to characterize the chemical constituents of the cladodes and fruits of Opuntia Mipa Alta. DPPH radical, hydroxyl radical(·OH) and superoxide anion radical(· ) scavenging ability were used to investigate the antioxidant activities of the cladodes and fruits of Opuntia Milpa Alta. RESULTS A total of 39 compounds were characterized from the cladodes and fruits of Opuntia Milpa Alta, including 22 phenolic compounds, 13 flavonoids and 4 other components. Both cladodes and fruits of Opuntia Milpa Alta had certain antioxidant capacity. The fruits of Opuntia Milpa Alta had better scavenging ability of DPPH than the cladodes, while the cladodes had a slightly stronger scavenging ability of ·OH than the fruits. The scavenging ability of · between cladode peels and fruit peels was not significantly different, and the juice had the strongest scavenging ability of · . The antioxidant activity of Opuntia Milpa Alta was closely related to its rich phenolic acids. CONCLUSION In this study, a rapid, accurate and reliable method is established to identify the chemical constituents of the cladodes and fruits of Opuntia Milpa Alta, which provides scientific basis for the comprehensive development and utilization of Opuntia Milpa Alta.

Objective: To study the application of urinary 5-aminolevulinic acid (5-ALA)detection in screening and identification of colorectal cancer and adenomatous polyps. Methods: The clinical data of 500 high-risk patients(including 22 cases with colorectal cancer, 134 cases with adenomatous polyps, and 344 cases with other patients) at the First Affiliated Hospital of Hebei North University from January 2018 to October 2018 were collected. High performance liquid chromatography(HPLC) was used to detect urinary 5-ALA and fecal occult blood test was used to detect faeces. Sensitivity and specificity of two methods was compared. At the same time, urine samples of 431 cases(including 22 cases with colorectal cancer, 134 cases with adenomatous polyps and 275 cases with colorectal normal mucosa)were collected, and the difference of the content of urinary 5-ALA among three groups was compared. Results: The sensitivity of urinary 5-ALA for the colorectal cancer screening was74.9%, and the specificity was 72.5%. The sensitivity of urinary 5-ALA for the adenomatous polyps screening was 70.1%, and the specificity was75.0%. The sensitivity of fecal occult blood test for the colorectal cancer screening was 63.6%, and the specificity was 62.1%. The sensitivity of fecal occult blood test for the adenomatous polyps screening was 42.3%, and the specificity was 62.5%. The content of urinary 5-ALA of the colorectal cancer group [(9.35 ± 0.46) μmol/g] was significantly higher than that of the adenomatous polyps group [(7.24 ± 0.64) μmol/g] (P < 0.05) and normal colorectal mucosa group [(3.12 ± 0.24) μmol/g] (P < 0.05), and the content of urinary 5-ALA of the adenomatous polyps group was significantly higher than that of colorectal normal mucosa group (P < 0.05). Conclusions For screening of colorectal cancer and adenomatous polyps, the content of urinary 5-ALA by HPLC is better than fecal occult blood test, and this approach can do great help to identify colorectal cancer, adenomatous polyps and normal colorectal mucosa.