ObjectiveTo evaluate the clinical efficacy of modified Houpo Dahuangtang in moderate and severe acute respiratory distress syndrome （ARDS） patients with phlegm-heat accumulation，and monitor the pulmonary ventilation changes of patients before and after treatment by electrical impedance tomography（EIT）. MethodThe 62 cases of moderate and severe ARDS patients with phlegm-heat accumulation who required mechanical ventilation in the department of intensive care unit （ICU） in Chongqing Hospital of Traditional Chinese Medicine from September 2021 to June 2022 were selected，and divided into an experimental group（31 cases）and a control group（31 cases）using a random number table. On the basis of regular Western medicine treatment，the experimental group received modified Houpo Dahuangtang and the control group received warm water by a nasogastric tube for seven days. The changes in the clinical efficacy of traditional Chinese medicine（TCM），the oxygenation index［arterial oxygen partial pressure （PaO₂）/fractional inspired oxygen（FiO₂），P/F］，lactic acid（Lac），acute physiology and chronic health evaluation Ⅱ（APACHE Ⅱ） score，compliance，plateau pressure，gas distribution parameters monitored by EIT（Z1，Z2，Z3 and Z4），inflammatory factors［interleukin-6 （IL-6），IL-10， tumor necrosis factor-α （TNF-α） and C-reactive protein （CRP）］ of both groups before and after treatment were recorded. Besides， the mechanical ventilation time， length of stay in ICU， 28-day mortality and incidence of adverse reactions（delirium，abdominal pain and diarrhea）in the two groups were also observed. ResultThere was no significant difference in the baseline indexes of patients in the two groups，and thus the two groups were comparable. After treatment for one week， the total effective rate for TCM syndromes in the experimental group was 90.30%（28/31）， higher than the 67.74%（21/31）in the control group（Z=-2.415，P<0.05）.Compared with the same group before treatment， the plateau pressure and Lac decreased （P<0.01）and the compliance and P/F increased （P<0.01） in experimental group， while the Lac decreased （P<0.05）and the P/F increased （P<0.05）， and the compliance and plateau pressure did not change significantly in the control group. After treatment，the plateau pressure and inflammatory factors in the experimental group were lower than those in the control group（P<0.05）， but the compliance and P/F in the experimental group were higher than those in the control group（P<0.05）， and the gas distribution parameters Z1，Z2，Z3，Z4，Z1+Z2，and Z3+Z4 monitored by EIT in the experimental group were all higher than those in the control group （P<0.05）. There was no significant difference in mechanical ventilation time， ICU hospitalization time， 28-day mortality， delirium， abdominal pain， diarrhea and other adverse reactions between the two groups. ConclusionModified Houpo Dahuangtang can significantly improve the P/F，pulmonary ventilation in gravity-dependent regions and pulmonary compliance，reduce the release of inflammatory factors in moderate and severe ARDS patients. Compared with conventional methods，EIT can timely monitor the pulmonary ventilation changes in ARDS patients，which suggests its clinical feasibility.

Objective:To investigate the safety and effectiveness of endoscopic retrograde cholangiopancreatography (ERCP) for the diagnosis and treatment of pediatric pancreaticobiliary maljunction (PBM).Methods:Data of 40 pediatric patients under 14 with PBM diagnosed and treated by ERCP at Department of Gastroenterology, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from November 2012 to September 2022 were collected. PBM types, ERCP-related diagnosis and treatment, adverse events and prognosis were retrospectively analyzed.Results:Nineteen cases were P-B type (joining of common bile duct with pancreatic duct), 17 were B-P type (joining of pancreatic duct with common bile duct), and 4 were complex type. Forty children with PBM underwent 50 ERCP-related operations, among which 48 procedures succeeded. One case failed during cannulation of ERCP, replaced by rendezvous-assisted endoscopic retrograde pancreatography (RV-ERP) afterwards. There were no serious postoperative adverse events such as bleeding, perforation or death. Thirty-four patients (85%) were followed up successfully, among which 14 underwent further surgery and 20 continued conservative treatment.Conclusion:ERCP is the golden standard to diagnose pediatric PBM, and it is also safe and effective treatment for PBM.

Objective:To investigate the expression of fatty acid desaturase 2 (FADS2) in psoriatic skin lesions, as well as its regulatory factors.Methods:FADS2 expression in psoriatic skin lesions was analyzed by using the dataset GDS4602 in Gene Expression Omnibus (GEO) database. Skin tissues were obtained from the back of 5 C57BL/6 mouse models of imiquimod-induced psoriasis, normal skin of 4 patients without psoriasis or other immune skin diseases, lesions of 4 patients with psoriasis before and after 10-week treatment with infliximab, as well as lesions of 3 patients with psoriasis before and after 12-week treatment with secukinumab in Shanghai Skin Disease Hospital. FADS2 expression was determined by both immunohistochemical staining and Western blot analysis in the epidermis of mouse skin tissues, and by immunohistochemical staining in that of human skin tissues. In vitro cultured human immortalized keratinocytes (HaCaT) were divided into several groups to be treated with 50 ng/ml tumor necrosis factor-α (TNF-α) alone for 0, 6, 12 and 24 hours respectively, 200 ng/ml interleukin-17A (IL-17A) alone for 0, 6 and 12 hours respectively, or treated with 50 ng/ml TNF-α and 5 μmol/L BAY 11-7082 (a nuclear factor-κB pathway inhibitor) for 6 hours (TNF-α+ BAY 11-7082 6 h group) , and the cells receiving normal culture served as the control group. After the above treatment, real-time fluorescence-based quantitative PCR (qPCR) and Western blot analysis were conducted to determine the mRNA and protein expression of FADS2 respectively. Statistical analysis was carried out by using one-way analysis of variance and t test. Results:Analysis of the dataset GDS4602 showed that the FADS2 mRNA expression was significantly lower in the lesional and non-lesional skin tissues from the patients with psoriasis (0.656 ± 0.475, 1.503 ± 1.062, respectively) than in the normal skin tissues (2.035 ± 1.226; F = 55.17, 3.07, P < 0.001, = 0.012, respectively) , and was significantly lower in the lesional skin tissues than in the non-lesional skin tissues from the patients with psoriasis ( F = 26.27, P < 0.001) . Western blot analysis and immunohistochemical staining both showed significantly decreased FADS2 protein expression in the mouse skin tissues in the imiquimod group (gray-value ratio: 0.463 ± 0.172; fluorescence intensity: 21.840 ± 3.125) compared with the normal control group (gray-value ratio: 1.000, t = 7.00, P = 0.002; fluorescence intensity: 30.720 ± 6.850, t = 3.15, P = 0.035) . Compared with the skin lesions before treatment, the FADS2 protein expression significantly increased in the skin lesions from the patients with psoriasis after 10-week treatment with infliximab (43.775± 3.342 vs. 27.950 ±1.218, t = -6.95, P = 0.006) , but was not significantly changed in the skin lesions from the patients with psoriasis after 12-week treatment with secukinumab (28.667 ± 3.402 vs. 31.933 ± 2.987, t = 2.72, P = 0.113) . qPCR revealed that the FADS2 mRNA expression significantly decreased in HaCaT cells in the TNF-α 6 h group and TNF-α 12 h group compared with the TNF-α 0 h group ( P = 0.002, 0.003, respectively) , while there was no significant change in the FADS2 mRNA expression in the IL-17A 6 h group and IL-17A 12 h group compared with the IL-17A 0 h group ( P = 0.849, 0.961, respectively) . The FADS2 mRNA expression significantly decreased in HaCaT cells in the TNF-α 6 h group (0.682 ± 0.132) compared with the control group (1.000, t = 4.82, P = 0.017) , but significantly increased in the TNF-α + BAY 11-7082 6 h group (1.541 ± 0.525) compared with the TNF-α 6 h group ( t = -3.58, P = 0.037) . Western blot analysis revealed significantly decreased FADS2 protein expression in HaCaT cells in the TNF-α 24 h group compared with the TNF-α 0 h group ( F = 6.24, P = 0.013) . Conclusion:FADS2 expression was downregulated in psoriatic lesions, which may be related to TNF-α.

Objective:To investigate changes in circadian gene cryptochrome 2 (CRY2) expression in mouse models of psoriasis and HaCaT cells, and to explore underlying mechanisms.Methods:Imiquimod-induced mouse model experiment: 12 C57BL/6 female mice were randomly and equally divided into imiquimod group receiving topical imiquimod treatment for 5 consecutive days and control group receiving no treatment; these mice were sacrificed on day 6, skin tissues were resected from the back of mice, and immunofluorescence staining was performed to determine the CRY2 expression in the epidermis. HaCaT cell transfection experiment: HaCaT cells with small interfering RNA (siRNA) -mediated knockdown of CRY2 served as siRNA-CRY2 group, and siRNA-NC group as control group; 5-ethynyl-2′-deoxyuridine (EdU) staining was performed to evaluate the proliferative activity of the HaCaT cells, real-time fluorescence-based quantitative PCR (qPCR) to determine the mRNA expression of chemokines in the HaCaT cells, and Western blot analysis to determine phosphorylation levels of extracellular signal-regulated kinase 1/2 (ERK1/2) . Tumor necrosis factor-α (TNF-α) -stimulated animal and cell experiments: 12 C57BL/6 female mice were randomly and equally divided into TNF-α group subcutaneously injected with TNF-α solution in the ear for 6 days, and phosphate buffered saline (PBS) group subcutaneously injected with the same amount of PBS; the mice were sacrificed on day 7, skin tissues were resected from the ear of mice, and immunofluorescence staining was conducted to determine the CRY2 expression in the epidermis; CRY2-knockdown HaCaT cells stimulated with 50 ng/ml TNF-α for 12 hours served as siRNA-CRY2 + TNF-α group, and siRNA-NC + TNF-α group as control group; qPCR was performed to determine the mRNA expression of chemokines in HaCaT cells in the above groups. Statistical analysis was carried out by using two-independent-sample t test. Results:Immunofluorescence staining showed that the CRY2 protein expression was significantly lower in the mouse dorsal epidermis in the imiquimod group (0.94 ± 0.23) than in the control group (2.30 ± 0.25, t = 3.99, P = 0.016) . Compared with the siRNA-NC group, the siRNA-CRY2 group showed significantly increased proportions of EdU-positive cells (48.13% ± 10.97% vs. 38.23% ± 0.81%, t = 5.00, P = 0.007) , mRNA expression levels of chemokines CXCL1 and CXCL8, as well as significantly increased phosphorylated (p) -ERK1/2 protein expression levels (all P < 0.05) , while there were no significant differences in the CCL20 mRNA expression or ERK1/2 protein expression between the two groups (both P > 0.05) . Immunofluorescence staining showed significantly decreased CRY2 protein expression level in the mouse ear epidermis in the TNF-α group (0.37 ± 0.34) compared with the PBS group (2.04 ± 0.17, t = 4.38, P = 0.012) ; the relative mRNA expression levels of chemokines CXCL1, CXCL8, and CCL20 in HaCaT cells were significantly higher in the siRNA-CRY2 + TNF-α group than in the siRNA-NC + TNF-α group (all P < 0.05) . Conclusion:CRY2 was markedly underexpressed in psoriasis, which might promote the proliferation of keratinocytes and expression of chemokines CXCL1, CXCL8 and CCL20, and TNF-α might be an upstream cytokine that could downregulate CRY2 expression.

Objective:To investigate the efficacy and safety of a new China-made direct visualization system of peroral cholangiopancreatography in the diagnosis and treatment of biliopancreatic diseases.Methods:Clinical data of 37 patients who underwent endoscopic examination through the direct visualization system of peroral cholangiopancreatography at Digestive Endoscopy Center of Nanjing Drum Tower Hospital from April 2020 to June 2021 were retrospectively analyzed. Technical success rate and complications were analyzed.Results:The examination was completed in 37 patients through the system. The technical success rate was 100.0%. The nature of biliary stricture was confirmed in 24 cases, presenece or absence of bleeding or residual stones in the bile duct was confirmed in 6 cases, neoplasm or residual stones in the pancreatic duct was determined in 2 cases, biliary stricture was passed assisted with visualized guidewire in 2 cases, and lithotripsy was performed assisted with biliary laser in 3 cases. Nine patients were pathologically diagnosed as having malignant biliary stricture, and 8 of them were confirmed malignant by the system. Drainage was performed in 34 cases after the examination. There were 3 cases of cholangitis, 4 cases of bacteremia and 2 cases of postoperative pancreatitis after the operation, which were relieved in a short time after conservative treatment. Bleeding occurred in 1 case which was improved after two times of endoscopic hemostasis.Conclusion:The new direct visualization system of peroral cholangiopancreatography is safe and effective in the diagnosis and treatment of biliopancreatic diseases.

Objective:To explore the predictive value of peripheral blood CD34-positive cell count for the stem cell mobilization effect of plerixafor in patients with multiple myeloma (MM).Methods:The clinical data of 12 MM patients who used plerixafor for stem cell mobilization in the First Affiliated Hospital of Guangxi Medical University from December 2019 to February 2021 were retrospectively analyzed. The changes of peripheral blood CD34-positive cell count and the collection status of stem cell in all patients before and after the mobilization of plerixafor were analyzed.Results:Twelve patients were included in this study. These patients were in international staging system (ISS) stage Ⅱ-Ⅲ, and the induction therapy was mainly VRD regimen. The CD34-positive cell count was increased after the use of plerixafor in all patients no matter which mobilization strategies were used before plerixafor. The CD34-positive cell count was 3.63/μl (0.72-13.53/μl) and 32.11/μl (8.52-53.68/μl) before and after the use of plerixafor, and the difference was statistically significant ( Z = -0.40, P<0.001); the median increasing time was 11.50 times (1.61-23.71 times). The mobilization failure occurred in 1 patient. The CD34-positive cell count in his blood was less than 1/μl before the use of plerixafor; though increased 11.83 times after the use of plerixafor, the CD34-positive cell count was still less than 10/μl. Pearson analysis showed that among the patients with CD34-positive cell count less than 4/μl before the use of plerixafor, there was a positive correlation in peripheral blood CD34-positive cell count before and after the use of plerixafor ( r = 0.80, P = 0.032). Conclusions:The peripheral blood CD34-positive cell count has a certain predictive value for the stem cell mobilization effect of plerixafor in MM patients.

Objective:To evaluate the application value of small IT knife pre-cut in assistance to duodenal papillary cannulation compared with conventional guidewire cannulation.Methods:A retrospective analysis was performed on the data of 90 patients with choledocholithiasis including 52 patients with small IT knife pre-cut assisted intubation (small IT knife group) and 38 patients with conventional guidewire intubation (conventional group) in endoscopic retrograde cholangiopancreatography (ERCP) from May 2016 to July 2019 in the digestive endoscopy center of Nanjing Drum Tower Hospital. The basic data, curative effect and complications of the two groups were collected and compared.Results:There was statistically significant difference in gender composition between the small IT knife group and the conventional group ( χ2=5.679, P=0.017), but no significant difference in other baseline data between the two groups (all P>0.05). The median intubation time of the small IT knife group was significantly shorter than that of the conventional group (141.5 s VS 270.0 s, Z=1 268.0, P=0.022). There were no significant differences in the success rate of intubation [98.1% (51/52) VS 94.7% (36/38), χ2=0.760, P=0.571], the incidence of intraoperative bleeding [15.4% (8/52) VS 7.9% (3/38), χ2=1.148, P=0.345], postoperative pancreatitis [5.8% (3/52) VS 7.9% (3/38), χ2=0.159, P=0.694], and postoperative cholangitis [1.9% (1/52) VS 5.3% (2/38), χ2=0.760, P=0.571] between the two groups. No perforation occurred in the two groups. After stratifying according to the operator′s proficiency, the median intubation time was significantly different between the small IT knife expert group and the conventional expert group (116.0 s VS 258.0 s, Z=276.0, P=0.038), while there was no significant difference in the intubation time among other groups (all P>0.05). Conclusion:The small IT knife is safe and effective to pre-cut and assist intubation in ERCP, and it may shorten the intubation time.

Objective: To investigate the prerequisites for endoscopists, who were chosen to receive endoscopic submucosal dissection (ESD) training. Methods: A total of 41 trainees, who attended ESD training in the endoscopic center of Nanjing Drum Tower Hospital from January 2017 to June 2018 were enrolled in the study. The general information of the subjects were collected, including name, age, gender, the number of gastroscopy and colonoscopy independently performed before training, the independent usage of narrow band image (NBI), magnifying endoscopy (ME), endoscopic mucosal resection (EMR), ESD, endoscopic ultrasonography (EUS), and endoscopic retrograde cholangiopancreatography (ERCP) before training. And then every trainee independently finished four in vitro experiments of pig esophagus ESD. The specimen area and operating time were recorded, and the operating speed was calculated. Linear regression analysis was used to analyze the affecting factors of operating speed of ESD. Results: Among the 41 trainees, 26 were male and 15 were female, with age of 36.07±4.44 years. The specimen area, operating time and operating speed of pig esophagus ESD was 4.67±1.61 cm^2, 24.54±5.97 min, and 0.19±0.05 cm²/min, respectively. Univariate linear regression analysis showed that the number of gastroscopy (n>5 000, P=0.001 8) and colonoscopy (n>3 000, P=0.000 1), the detect number of early cancer in upper digestive tract (n>30, P=0.000 3) and lower digestive tract (n>10, P=0.019 7), and the usage of ME (P=0.047 8), EMR (P=0.019 6) and ESD (P=0.000 3) before training were statistically correlated with the operating speed of ESD. Carrying out NBI (P=0.532 9), ERCP (P=0.500 7) and EUS (P=0.766 8) before training were not statistically correlated with the operating speed of ESD. The operating speed of ESD was negatively correlated with the perforation rate of ESD (P<0.000 1). According to multivariable linear regression model, the usage of EMR (P=0.029) and ESD (P=0.034) were statistically correlated with the operating speed of ESD. Conclusion ESD trainees, who have the number of gastroscopy more than 5 000, the number of colonoscopy more than 3 000, the detect number of early cancer more than 30 in upper digestive tract and 10 in lower digestive tract, and the usage of ME, EMR and ESD before training, can get a better training effect. The study provides a theoretical basis for selecting appropriate ESD trainees.

Objective: To investigate the therapeutic effect of dendritic cell-induced killer cells (DC-CIK) combined with 5-fluorouracil (5-FU) and loaded with CD133+ HT-29 cell lysate or RNA on mice bearing colon cancer HT-29 cell transplanted tumor, and to explore the underlying mechanism. Methods: Colon cancer xenograft model was established in BALB /c nude mice by using human colon cancer HT-29 cells at logarithmic growth phase; Antigen-free DC-CIK, 5-FU+DC-CIK, R+DC-CIK (loaded with total RNA of CD133+ cells), L+DC-CIK (loaded with CD133+ cell lysate), 5-FU and normal saline were respectively injected into transplanted mice, and the treatment efficacies on the growth of transplanted tumor in each group after three treatment cycles were observed, and the tumor growth curve was drawn. The nude mice were sacrificed by cervical dislocation and the tumor volume and body weight were measured. qPCR was used to detect the expression of AKT mRNA in transplanted tumor tissue, and WB was used to detect the expression of phosphorylated AKT protein. Results: After treatment, the body mass of nude mice in R+DC-CIK group, L+DC-CIK group and 5-FU+DC-CIK group increased steadily, while the body mass of nude mice in DC-CIK group and 5-FU group decreased gradually; the tumor growth speed of nude mice in R+DC-CIK group, 5-FU+DC-CIK group and L+DC-CIK group was significantly slower than that of the control group (P<0.05). Compared with 5-FU and DC-CIK alone, the combined treatment with loaded lysate/RNA had more sig nificant effect on mRNA and protein expressions of AKT(P<0.05). Conclusion: The effect of DC-CIKwithdifferentloadingoritscombinationwith5-FUisbetterthanthatofchemotherapy alone. One of the mechanisms is related to the down-regulation ofAKT level.

Objective To investigate the effect of human umbilical cord mesenchymal stem cells (HUC-MSCs) on CD4+T cells in liver after hepatic ischemia-reperfusion injury (HIRI) in mice. Methods Two hundred and twenty-five mice were randomly divided into sham group, control group and MSC group, with 75 mice in each group. HIRI model mice were used in MSC group and control group. HUC-MSCs were injected in MSC group through inferior vena cava. Normal saline was injected in control group through inferior vena cava. Only laparotomy and abdominal closure were performed in sham group without blood vessel clipping. At 6, 12 and 24 h after operation, 15 mice of each group were randomly selected to sample eyeball blood and liver tissues, and the 30 mice left in each group were used to extract intrahepatic mononuclear cells. The number of intrahepatic mononuclear cells, percentage, number and positive rate of CD4+T cells in the mice of various groups at different time points were compared. The content of interleukin (IL)-17 in serum and liver tissue as well as expression levels of costimulatory molecules B7-1 and B7-2 messenger RNA (mRNA) in liver tissues of the mice at different time points were compared. Results At 12 and 24 h after operation, the number of intrahepatic mononuclear cells of control group was significantly higher than that of sham group, while the number of intrahepatic mononuclear cells of MSC group was significantly lower than that of control group (P<0.01-0.05). At 6, 12 and 24 h after operation, the percentage, number and positive rate of CD4+T cells of control group were significantly higher than those of sham group (all P<0.01), while the percentage of CD4+T cells of MSC group was significantly lower than that of control group (P<0.01-0.05). At 12 and 24 h after operation, the number and positive rate of CD4+T cells of MSC group were significantly lower than those of control group (P<0.01-0.05). At 6, 12 and 24 h after operation, the IL-17 contents in serum and liver tissues of control group were higher than those of sham group (all P<0.01), while the IL-17 contents in serum and liver tissues of MSC group were lower than those of control group (all P<0.01). At 6 h after operation, the mRNA expression level of B7-2 of control group was higher than that of sham group (P<0.05). At 12 and 24 h after operation, the mRNA expression levels of B7-1 and B7-2 of control group were higher than those of sham group (all P<0.01), while the mRNA expression levels of B7-1 and B7-2 of MSC group were lower than those of control group (all P<0.01). Conclusions HUC-MSCs inhibits the number of CD4+T cells and the secretion of IL-17 in liver after HIRI, as well as decreases the number of intrahepatic mononuclear cells and the mRNA expression of B7-1 and B7-2, thereby alleviating HIRI.