Objective This study aimed to investigate the glycosyltransferase gene DsUGT11 involved in the biosynthesis of flavonoids in Desmodium styracifolia,and to analyze the function of its encoding protein by bioinformatic tools,gene cloning,prokaryotic expression,and other technologies.Methods The sequence characteristics and potential biological functions of DsUGT11 were analyzed and predicted by bioinformatics analysis,respectively.Total RNA was extracted from fresh leaves and reverse transcribed into cDNA,from which DsUGT11 gene was successfully amplified and cloned.Heterologous expressed protein was induced and purified,followed by functional characterization using enzymatic reaction in vitro.Results A candidate glycosyltransferase gene,designated as DsUGT11,was identified from the transcriptome data of D.styracifolia.The length of the open reading frame of DsUGT11 is 1426 bp,and the molecular weight of its encoding protein is expected to be 52.14 KDa.By bioinformatic analysis,DsUGT11 was found to harvest a conserved motif of"PSPG"that is unique to the UGT family.Moreover,DsUGT11 was successfully amplified and cloned using the prokaryotic expression vector pMALc5X.Recombinant protein was induced and purified subsequently.Next,the purified protein was used to perform the enzymatic reaction in vitro,the result of which suggested that DsUGT11 was able to catalyze the conversion of 2-OH-naringenin and UDP-glucose into three different compounds,one of which was authenticated as apigenin-7-O-β-D-glucoside(also known as Apigetrin),with two others unknown.Conclusion In this study,the DsUGT11 gene was identified and cloned,whose encoding protein is a flavone-oxyglycosyltransferase catalyzing the conversion of 2-OH-naringenin and UDP-glucose into three different compounds including Apigetrin.

Objective To investigate the effects of interleukin-9 (IL-9) in the activation of hepatic stellate cells (HSCs) in mice infected with Schistosoma japonicum. Methods Primary HSCs were isolated from mice 7 weeks post-infection with S. japonicum using the in situ liver perfusion and density gradient centrifugation, and cultured in vitro. HSCs were randomly assigned to the PBS control group and IL-9 stimulation group (stimulation with 20 ng/mL IL-9). HSCs were harvested 48 h and 72 h poststimulation, and the expression of α-smooth muscle actin (α-SMA), type I collagen (Col I) and type III collagen (Col III) was determined in HSCs using Western blotting. Results Following stimulation with 20 ng/mL IL-9 for 48 h, the expression of α-SMA [(0.87 ± 0.02) vs. (0.69 ± 0.01); t = 17.39, P < 0.01], Col I [(0.74 ± 0.02) vs. (0.65 ± 0.01); t = 9.56, P < 0.01] and Col III [(0.94 ±0.04) vs. (0.75 ± 0.03); t = 6.15, P < 0.01] was significantly greater in HSCs in the IL-9 stimulation group than in the PBS control group. Following stimulation with 20 ng/mL IL-9 for 72 h, the expression of α-SMA was significantly greater in HSCs in the IL-9 stimulation group than in the PBS control group[(0.76 ± 0.02) vs. (0.58 ± 0.02); t = 12.52, P < 0.01]; however, there were no significant differences between the two groups in terms of Col I [(0.68 ± 0.02) vs. (0.66 ± 0.02); t = 1.15, P > 0.05] or Col III expression [(0.75 ± 0.01) vs. (0.72 ± 0.02); t = 2.22, P > 0.05]. Conclusions IL-9 promotes the activation of HSCs in mice infected with S. japonicum.

OBJECTIVES: To study the clinical features of intestinal polyps and the risk factors for secondary intussusception in children. METHODS: A retrospective analysis was performed for the medical data of 2 669 children with intestinal polyps. According to the presence or absence of secondary intussusception, they were divided into two groups: intussusception (n=346) and non-intussusception (n=2 323). Related medical data were compared between the two groups. The multivariate logistic regression analysis was used to identify the risk factors for secondary intussusception. RESULTS: Among the children with intestinal polyps, 62.42% were preschool children, and the male/female ratio was 2.08∶1; 92.66% had hematochezia as disease onset, and 94.34% had left colonic polyps and rectal polyps. There were 346 cases of secondary intussusception, with an incidence rate of 12.96% (346/2 669). Large polyps (OR=1.644, P<0.001), multiple polyps (≥2) (OR=6.034, P<0.001), and lobulated polyps (OR=93.801, P<0.001) were the risk factors for secondary intussusception. CONCLUSIONS Intestinal polyps in children often occur in preschool age, mostly in boys, and most of the children have hematochezia as disease onset, with the predilection sites of the left colon and the rectum. Larger polyps, multiple polyps, and lobulated polyps may increase the risk of secondary intussusception, and endoscopic intervention is needed as early as possible to improve prognosis.
Child, Preschool ; Female ; Gastrointestinal Hemorrhage ; Humans ; Intestinal Polyps/complications* ; Intussusception/complications* ; Male ; Retrospective Studies ; Risk Factors

Objective: To investigate the safety and feasibility of laparoscopic double-flap technique (Kamikawa) in digestive tract reconstruction after proximal gastrectomy for esophagogastric junction (EGJ) leiomyoma and gastrointestinal stromal tumor (GIST) with the maximum diameter >5 cm. Methods: A descriptive case-series study was used to retrospectively analyze the data of patients with EGJ leiomyoma and GIST undergoing laparoscopic-assisted proximal gastrectomy and double-flap technique (Kamikawa) at the Department of Gastrointestinal Surgery, Guangdong Hospital of Traditional Chinese Medicine from September 2017 to March 2019. All the tumors invaded the cardia dentate line, and the maximum diameter was >5 cm. After the exclusion of patients requiring emergency surgery and complicating with severe cardiopulmonary diseases, a total of 4 patients, including 3 males and 1 female with age of 29-49 years, were included in this study. After laparoscopic-assisted proximal gastrectomy, the residual stomach was pulled out of the abdominal cavity and marked with methylene blue at the proximal end 3~4 cm from the anterior wall of the residual stomach in the shape of "H". The gastric wall plasma muscular layer was cut along the "H" shape, and the space between the submucosa and the muscular layer was separated to both sides along the longitudinal incision line to make the seromuscular flap. The residual stomach was put back into the abdominal cavity. Under laparoscopy, 4 stitches were intermittently sutured at the upside of "H" shape and 4-5 cm from the posterior wall of the esophageal stump. The stump of the esophagus was cut open, and the submucosa and mucosa were cut under the "H" shape to enter the gastric cavity. The posterior wall of the esophageal stump was sutured continuously with the gastric stump mucosa and submucosa under laparoscopy. The anterior wall of the esophageal stump was sutured continuously with the whole layer of the residual stomach. The anterior wall of the stomach was sutured to cover the esophagus. The anterior gastric muscle flap was sutured and embedded in the esophagus to complete the reconstruction of digestive tract. The morbidity of intraoperative complications and postoperative reflux esophagitis and anastomosis-related complications were observed. Results: All the 4 patients completed the operation successfully, and there was no conversion to laparotomy. The median operative time was 239 (192-261) minutes, the median Kamikawa anastomosis time was 149 (102-163) minutes, and the median intraoperative blood loss was 35 (20-200) ml. The abdominal drainage tube and gastric tube were removed, and the fluid diet was resumed on the first day after surgery in all the 4 patients. The median postoperative hospitalization time was 6 (6-8) days. Postoperative pathology revealed 3 leiomyomas and 1 GIST. There were no postoperative complications such as anastomotic leakage or stenosis, and no reflux symptoms were observed. The median follow-up time was 22 (11-29) months after the operation, and no reflux esophagitis occurred in any of the 4 patients by gastroscopy. Conclusion: For >5 cm EGJ leiomyoma or GIST, double-flap technique (Kamikawa) used for digestive tract reconstruction after proximal gastrectomy is safe and feasible.
Adult ; Anastomosis, Surgical/methods* ; Esophagogastric Junction/surgery* ; Esophagus/surgery* ; Feasibility Studies ; Female ; Gastrectomy/methods* ; Gastrointestinal Stromal Tumors/surgery* ; Humans ; Laparoscopy ; Leiomyoma/surgery* ; Male ; Middle Aged ; Retrospective Studies ; Stomach/surgery* ; Stomach Neoplasms/surgery* ; Surgical Flaps ; Treatment Outcome

Traditional Chinese medicine(TCM) injections boast a definite efficacy and have been widely used in clinic. However, the problems in medication safety have been attracted increasing attention. Pharmacokinetics is of significance to guiding TCM injection administration regimen design and improving safety and effectiveness in clinical use. In recent years, with the improvement of ideas, technology and methods of TCM studies, the pharmacokinetic studies of TCM injections have been broadly performed, with a notable progress. This paper reviewed the advance in pharmacokinetics studies of TCM injections in recent ten years, which mainly focused on pre-clinical concentration-time course, distribution, metabolism and excretion in vivo based on analysis techniques, pharmacokinetic interactions of constitutes, impact of pathological state, pharmacokinetic interactions between TCM injection and chemical drugs, and clinical pharmacokinetics studies of TCM injections, in the expectation of providing reference for studies on quality control, product development and rational clinical use of TCM injections.
Drugs, Chinese Herbal ; Injections ; Medicine, Chinese Traditional ; Quality Control

ObjectiveRecent studies revealed that the transcription factor EB (TFEB) plays an important role in regulating autophagy, reducing intracellular lipids, and inhibiting atherosclerosis. This study aims to explore the effects of atorvastatin (ATV) on autophagy and cholesterol levels of foam cells by activating TFEB.MethodsHuman mononuclear cell line THP-1 was cultured in vitro and induced to differentiate into macrophages using phorbol ester. Oxidized low-density lipoprotein (oxLDL) was added to macrophages, which were induced for 48 hours to establish a foam cell model. The experiment was divided into four groups: blank group, model group (oxLDL group), oxLDL+ Chloroquine (CQ) group, oxLDL+ ATV group, and oxLDL+CQ+ ATV group. Cells in each group were treated with drugs for 48 h. The toxicity of ATV and chloroquine on the cells was detected by the CCK8 method. Oil red O staining was used to test the level of lipid droplets. Oxidase method was used to detect levels of intracellular free cholesterol (FC), total cholesterol (TC) and others related to. Cholesterol efflux fluorescence analysis was used to determine the cholesterol efflux rate of the cells. Expression of I, P62, TFEB, LAMP 1 protein was determined by Western blot.ResultsThe results of the CCK8 method showed that the cell survival rate decreased significantly with the increase of ATV and CQ concentrations (P<0.01). Compared to the blank group, the levels of lipid droplets, FC, TC, and CE/TC in the model group significantly increased (P<0.05), and the cholesterol outflow rate significantly decreased (P<0.05). Compared to the model group, the intracellular lipid droplets, FC, TC, and CE/TC levels in the oxLDL+CQ group elevated significantly (P<0.05), while the cholesterol outflow rate decreased significantly (P<0.05). The intracellular lipid droplets, FC, TC, and CE/TC contents in the oxLDL+ATV group decreased significantly (P<0.05), and the cholesterol outflow rate increased significantly (P<0.05). Compared to the oxLDL+CQ group, the intracellular lipid droplets, FC, TC, and CE/TC content in the oxLDL+CQ+ATV group decreased significantly (P<0.05), while the cholesterol outflow rate increased significantly (P<0.05). Western blotting results showed that protein expression levels of LC3II/I, P62 and TFEB were decreased in the model histone in comparison to the blank group (P<0.05). Compared to the expression levels of LC3II/I, P62, TFEB and LAMP1 ((1.006±0.052), (0.183±0.013), (0.333±0.020), and (0.957±0.026)) in the model group, the expression levels of oxLDL+CQ histamine ((1.594±0.017), (0.257±0.006), (0.477±0.024), and (0.957±0.026)) were significantly higher (P<0.05).The protein expression levels of LC3II/I, TFEB and LAMP1 in oxLDL+ATV group ((1.146±0.060), (0.540±0.031), and (1.027±0.054)) were increased, while the protein expression levels of P62 (0.115±0.009) were decreased (P<0.05). Compared to LC3II/I and P62 in oxLDL+CQ group, the expression level of oxLDL+CQ+ATV histone was decreased ((1.419±0.036) and (0.165±0.006)), and the difference was statistically significant (P< 0.05).ConclusionATV can increase the cholesterol outflow rate of macrophages, reduce the level of cholesterol and lipid droplets in macrophages, and reduce the formation of foam cells. The mechanisms behind this are still unknown, which may be related to the activation of TFEB by ATV and influencing the process of autophagy.

Objective: To investigate the influence of the degrees of intravesical prostatic protrusion (IPP) on the recovery of urinary continence after radical prostatectomy. METHODS: We retrospectively analyzed the clinical data on 212 patients diagnosed with prostate cancer by biopsy and treated by laparoscopic radical prostatectomy by the same surgeon. Based on the degrees of IPP measured by MRI, we divided the patients into an IPP ≤ 10 mm group (n = 146) and an IPP > 10 mm group (n = 66) and determined the factors influencing the recovery of urinary continence by univariate and multivariate logistic regression analyses. RESULTS: At 1, 3, 6 and 12 months after surgery, the urinary continence rates of the patients were 32.5%, 50.5%, 82.1% and 91%, respectively. Univariate analysis indicated that the factors influencing the recovery of urinary continence included IPP, body mass index (BMI), bladder neck preservation (BNP), neurovascular bundle preservation (NVBP) and clinical tumor (T) stage at 3 months (P < 0.05 or P < 0.01), age, IPP, BMI, BNP and clinical T stage at 6 months (P < 0.05 or P < 0.01), and age, IPP, BMI, BNP, NVBP and clinical T stage at 12 months (P < 0.05), while multivariate logistic regression analysis showed the independent influencing factors to be IPP > 10 mm (P < 0.001), BMI ≥ 25 kg/m2 (P = 0.004) and BNP (P = 0.032) at 3 months, and IPP and BMI at 6 months (both P < 0.01) and 12 months (P < 0.01 and P = 0.033). CONCLUSIONS IPP > 10 mm and BMI ≥ 25 kg/m2 are independent factors influencing the long-term recovery of urinary continence after radical prostatectomy.

To detect the expression level of ICOS on Th9 cells in mice infected with Schistosoma japonicum, and investigate the relation between ICOS signaling and Th9 cell polarization.Twenty-five mice with S. japonicum infection were used as models. IL-9⁺cells in CD4⁺ T cells and ICOS⁺ cells in Th9 cells of the mice were detected by flow cytometry 0, 4, 7, 9 weeks and 12 weeks after the infection.Compared with that 0 week after the infection, the proportion of Th9 cells in CD4⁺ T cells of the mice significantly increased 4, 7, 9, 12 weeks after the infection (all P < 0.05), and the proportion of ICOS⁺ cells in Th9 cells also markedly improved (P < 0.05).In S. japonicum infection, the ICOS signaling may have a regulatory effect on Th9 cell polarization.

Objective To investigate the effect of different concentrations of magnesium-calcium alloy extract on the expression of matrix metalloproteinase-9 (MMP9) and tissue inhibitor of metalloproteinase-3 (TIMP3) in human colonic epithelial NCM460 cells.Methods The different concentrations of extracts (the volume fraction was 10％,50％ and 100％ respectively) were made with magnesium-calcium alloy.The 5 × 106 L-1 NCM460 suspension was randomly divided into control group,experimental group 1,experimental group 2 and experimental group 3.The cells in the control group were cultured by 2 000 μL high glucose Dulbecco's modified Eagle's medium (containing 10％ volume fraction of fetal bovine serum).The cells in the experimental group 1,2 and 3 were cultured by 2 000 μL magnesium-calcium alloy extract with volume fraction of 10％,50％ and 100％ respectively.The expressions of MMP9 and TIMP3 mRNA in NCM460 cells was detected by real-time fluorescence quantitative polymerase chain reaction,and the expression of MMP9 and TIMP3 protein in NCM460 cells was detected by Western blot at after one,three and five days of cultivation respectively.Results The expression of MMP9 mRNA and TIMP3 mRNA in NCM460 cells of the experimental group 1,2 and 3 was significantly lower than that in the control group after one day of cultivation (P ＜ 0.05).After three and five days of cultivation,the expression of MMP9 mRNA in NCM460 cells of the experimental group 1 was significantly lower than that in the control group (P ＜ 0.05),but the expression of MMP9 mRNA in the NCM460 cells of the experimental group 2 and 3 was significantly higher than that in the control group and the experimental group 1 (P ＜ 0.05).The expression of MMP9 mRNA in NCM460 cells of the experimental group 3 was significantly higher than that in the experimental group 2 after five days of cultivation (P ＜ 0.05).The expression of MMP9 mRNA in NCM460 cells of the experimental group 1,2 and 3 after three and five days of cultivation was significantly higher than that after one day of cultivation(P ＜ 0.05).There was no significant difference in the expression of MMP9 mRNA in NCM460 cells of the experimental group 1 between three and five days of cultivation (P ＞ 0.05).The expression of MMP9 mRNA in NCM460 cells of the experimental group 2 and 3 after five days of cultivation was significantly higher than that after three days of cultivation(P ＜ 0.05).The expression of TIMP3 mRNA in NCM460 cells of the experimental group 2 and 3 was significantly higher than that in the experimental group 1 after one day of euhivation (P ＜ 0.05).After three days of cultivation,the expression of TIMP3 mRNA in NCM460 cells of the experimental group 1,2 and 3 was significantly lower than that in the control group (P ＜ 0.05);the expression of TIMP3 mRNA in NCM460 cells of the experimental group 2 was significantly lower than that in the experimental group 1 and 3 (P ＜ 0.05).After five days of cultivation,the expression of TIMP3 mRNA in NCM460 cells of the experimental group 1,2 and 3 was significantly higher than that in the control group (P ＜ 0.05).The expression of TIMP3 mRNA in NCM460 cells after three and five days of cultivation was significantly higher than that after one day of cultivation (P ＜ 0.05),and the expression of TIMP3 after five days of cultivation was significantly higher than that after three days of cultivation in the experimental group 1 (P ＜ 0.05).The expression of TIMP3 mRNA in NCM460 cells after three days of cultivation was significantly lower than that after one day of cultivation (P ＜ 0.05),and the expression of TIMP3 after five days of cultivation was significantly higher than that after one and three days of cultivation in the experimental group 2 (P ＜ 0.05).The expression of TIMP3 mRNA in NCM460 cells after five days of cultivation was significantly higher than that after one and three days of cultivation in the experimental group 3 (P ＜ 0.05).After five days of cultivation,there was no significant difference in the expression of MMP9 protein in NCM460 cells between the experimental group 1 and control group (P ＞ 0.05),the expression of MMP9 protein in NCM460 cells of the experimental group 2 and 3 was significantly higher than that in the control group and the experimental group 1 (P ＜ 0.05),but there was no significant difference in the expression of MMP9 protein in NCM460 cells between the experimental group 2 and 3 (P ＞ 0.05).After five days of cultivation,the expression of TIMP3 protein in NCM460 cells of the experimental group 1,2 and 3 was significantly higher than that in the control group (P ＜0.05);but there was no significant difference in the expression of TIMP3 protein in NCM460 cells among the experimental group 1,2and 3 (P ＞ 0.05).Conclusions The high concentration of magnesium-calcium alloy extract has certain influence on the expression of MMP9 and TIMP3 gene in NCM460 cells,which may lead to the early inflammatory reaction,and the mechanism may be related to the calcium ion concentration in the extract.

Objective To evaluate the effect of the endotracheal tube (ETT) depth on the efficacy of bronchial blocker when used for one-lung ventilation.Methods Sixty patients of both sexes,aged 46-78 yr,weighing 48-85 kg,with body mass index＜30 kg/m2,scheduled for elective thoracotomy requiring one-lung ventilation,were divided into 2 groups (n =30 each) using a random number table:the distance between the tip of ETT and the carina was 3 cm group (group Ⅰ) and the ETT cuff was placed at 2 cm below the glottis group (group Ⅱ).The bronchial blocker was inserted under the guidance of a fiberoptic bronchoscope.The ETT depth,time to bronchial blocker position,development of bronchial blocker displacement and increase in airway peak pressure,degree of lung collapse,severity of postoperative tracheal mucous membrane injury and development of sore throat and hoarseness were recorded.Results Compared with group Ⅰ,the ETT depth was significantly shallower,the time to bronchial blocker position was shortened,postoperative tracheal mucous membrane injury was attenuated (P＜0.05 or 0.01),and no significant change was found in the incidence of bronchial blocker displacement and increase in airway peak pressure,degree of lung collapse or incidence of sore throat and hoarseness in group Ⅱ (P＞0.05).Conclusion The ETT depth the ETT cuff placed at 2 cm below the glottis can improve the efficacy of bronchial blocker when used for one-lung ventilation.