ObjectiveMetabolomics was utilized to investigate the preventive effect of notoginseng total saponins（NTS） on aspirin（acetyl salicylic acid， ASA）-induced small bowel injury in rats. MethodFifty male SD rats were randomly divided into normal and model groups， NTS high-dose and low-dose groups（62.5， 31.25 mg·kg^-1）， and positive drug group（omeprazole 2.08 mg·kg^-1+rebamipide 31.25 mg·kg^-1）， with 10 rats in each group. Except for the normal group， rats in other groups were given ASA enteric-coated pellets 10.41 mg·kg^-1 daily to establish a small intestine injury model. On this basis， each medication group was gavaged daily with the corresponding dose of drug， and the normal group and the model group were gavaged with an equal amount of drinking water. Changes in body mass and fecal characteristics of rats were recorded and scored during the period. After 14 weeks of administration， small intestinal tissues of each group were taken for hematoxylin-eosin（HE） staining， scanning electron microscopy to observe the damage， and the apparent damage of small intestine was scored. Serum from rats in the normal group， the model group， and the NTS high-dose group was taken and analyzed for metabolomics by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS）， and the data were processed by multivariate statistical analysis， the potential biomarkers were screened by variable importance in the projection（VIP） value≥1.0， fold change（FC）≥1.5 or ≤0.6 and t-test P<0.05， and pathway enrichment analysis of differential metabolites was performed in conjunction with Human Metabolome Database（HMDB） and Kyoto Encyclopedia of Genes and Genomes（KEGG）. ResultAfter 14 weeks of administration， the average body mass gain of the model group was lower than that of the normal group， and the NTS high-dose group was close to that of the normal group. Compared with the normal group， the fecal character score of rats in the model group was significantly increased（P<0.05）， and compared with the model group， the scores of the positive drug group and the NTS high-dose group were reduced， but the difference was not statistically significant. HE staining and scanning electron microscopy results showed that NTS could significantly improve ASA-induced small intestinal injury， compared with the normal group， the small bowel injury score of the model group was significantly increased（P<0.01）， compared with the model group， the small bowel injury scores of the NTS low and high dose groups were significantly reduced（P<0.05， P<0.01）. Serum metabolomics screened a total of 75 differential metabolites between the normal group and the model group， of which 55 were up-regulated and 20 were down-regulated， 76 differential metabolites between the model group and the NTS groups， of which 14 were up-regulated and 62 were down-regulated. NTS could modulate three differential metabolites（salicylic acid， 3-hydroxybenzoic acid and 4-hydroxybenzoic acid）， which were involved in 3 metabolic pathways， namely， the bile secretion， the biosynthesis of folic acid， and the biosynthesis of phenylalanine， tyrosine and tryptophan. ConclusionNTS can prevent ASA-induced small bowel injury， and the underlying mechanism may be related to the regulation of bile secretion and amino acid metabolic pathways in rats.

Objective:To evaluate the effect of exogenous interleukin-18 (IL-18) binding protein (IL-18BP) on sepsis-associated lung injury and the role of NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasomes in rats.Methods:This study was performed in two parts. Experiment Ⅰ Thirty-six SPF-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 200 g, were allocated into 3 groups ( n=12 each) using a random number table method: sham operation group (S group), sepsis-associated lung injury group (SLI group), and sepsis-associated lung injury+ IL-18BP group (SI group). The rat model of sepsis-associated lung injury was developed using cecal ligation and puncture. The rats in group S underwent laparotomy without the subsequent ligation and puncture procedures. IL-18BP 1.5 mg/kg was intraperitoneally injected immediately after surgery in group SI. Experiment Ⅱ The aforementioned 36 rats were allocated into 3 groups ( n=12 each) using a random number table method: sepsis-associated lung injury + IL-18BP + vehicle group (SIV group), sepsis-associated lung injury + IL-18BP + MCC950 group (SIM group), and sepsis-associated lung injury + IL-18BP + nigericin group (SIN group). In SIV group, SIM group and SIN group, solvent, MCC950 (NLRP3 inhibitor, 5 mg/kg) and nigerin (NLRP3 agonist, 2 mg/kg) were intraperitoneally injected at 30 min before surgery respectively, and IL-18BP 1.5 mg/kg was intraperitoneally injected after operation. HE staining was used to observe the lung injury at 24 h after operation. The left lung tissues were obtained for calculation of the wet/dry weight (W/D) ratio. The concentrations of IL-18 and IL-18BP in broncho-alveolar lavage fluid (BALF) were determined by enzyme-linked immunosorbent assay. The percentage of NLRP3 positive cells in lung tissues was detected by immunofluorescence. The expression of IL-18, IL-18BP, cl-caspase-1 and gasdermin D (GSDMD) in lung tissues was detected by Western blot. Results:Experiment Ⅰ Compared with group S, the injury score of lung tissues, W/D ratio, concentrations of IL-18 and IL-18BP in BALF, and percentage of NLRP3 positive cells were significantly increased, and the expression of IL-18, IL-18BP, cl-caspase-1 and N-GSDMD was up-regulated in group SLI ( P<0.05). Compared with group SLI, the injury score of lung tissues, W/D ratio, concentrations of IL-18 and IL-18BP in BALF, and percentage of NLRP3 positive cells were significantly decreased, the expression of IL-18, cl-caspase-1 and N-GSDMD was down-regulated, and the expression of IL-18BP was up-regulated in group SI ( P<0.05). Experiment Ⅱ Compared with group SIV, the injury score of lung tissues, W/D ratio, concentrations of IL-18 in BALF, and percentage of NLRP3 positive cells were significantly decreased, and the expression of IL-18, cl-caspase-1 and N-GSDMD was down-regulated in group SIM, and the injury score of lung tissues, W/D ratio, concentrations of IL-18 in BALF, and percentage of NLRP3 positive cells were significantly increased, and the expression of IL-18, cl-caspase-1 and N-GSDMD was up-regulated in group SIN ( P<0.05). Conclusions:Exogenous IL-18BP can reduce sepsis-associated lung injury, and the mechanism is related to inhibition of activation of NLRP3 inflammasomes in rats.

BACKGROUND: Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome is a severe congenital disorder characterized by vaginal hypoplasia caused by dysplasia of the Müllerian duct. Patients with MRKH syndrome often require nonsurgical or surgical treatment to achieve satisfactory vaginal length and sexual outcomes. The extracellular matrix has been successfully used for vaginal reconstruction. METHODS: In this study, we developed a new biological material derived from porcine vagina (acellular vaginal matrix, AVM) to reconstruct the vagina in Bama miniature pigs. The histological characteristics and efficacy of acellularization of AVM were evaluated, and AVM was subsequently transplanted into Bama miniature pigs to reconstruct the vaginas. RESULTS: Macroscopic analysis showed that the neovaginas functioned well in all Bama miniature pigs with AVM implants. Histological analysis and electrophysiological evidence indicated that morphological and functional recovery was restored in normal vaginal tissues. Scanning electron microscopy showed that the neovaginas had mucosal folds characteristics of normal vagina. No significant differences were observed in the expression of CK14, HSP47, and a-actin between the neovaginas and normal vaginal tissues. However, the expression of estrogen receptor (ER) was significantly lower in the neovaginas than in normal vaginal tissues. In addition, AVM promoted the expression of b-catenin, c-Myc, and cyclin D1. These results suggest that AVM might promotes vaginal regeneration by activating the b-catenin/cMyc/cyclin D1 pathway. CONCLUSION This study reveals that porcine-derived AVM has potential application for vaginal regeneration.

Objective:To realize prokaryotic expression, purification and identification of 2019-novel Coronavirus (2019-nCoV) nucleocapsid protein (NP), and apply it to the serological diagnosis.Methods:The synthetic 2019-nCoV NP gene was cloned into the prokaryotic expression vector pET28a to construct expression plasmid, and then purified by Ni-chelating affinity. SDS-polyacrylamide gel electrophoresis (SDS-PAGE), indirect enzyme-linked immunosorbent assay (ELISA), Western blot (WB), and immunochromatography were used to test the purified protein. Indirect ELISA reaction conditions were optimized for serum antibody detection.Results:The relative molecular mass of recombinant NP was about 50×10 3 after SDS-PAGE electrophoresis, which was consistent with the expectation. Indirect ELISA and WB results showed that it could specifically bind to the serum of patients infected with 2019-nCoV. The detection limit of NP was 0.2 ng/ml by immunochromatography. The sera from 32 patients infected with 2019-nCoV and the control sera were detected by indirect ELISA, and the results showed that they were clearly clustered. Conclusions:Prokaryotic expression of 2019-nCoV NP has good immunogenicity and can be used for the development of serological diagnostic reagents.

Objective To explore the effect of Protectin DX(PDX) on acute liver injury(ALI) induced by sepsis in mice and the underlying mechanism. Methods Mice received cecum ligation and puncture(CLP) to induce sepsis-associated acute liver injury. Male C57BL/6 mice were randomly (random number) divided into 3 groups (n=10 each group): (1) sham group (S group), (2) CLP group and (3) CLP +PDX group (PDX group ). Mice in the PDX group were received PDX 1 μg (intraperitoneal injection). One hour after CLP operation, mice in the S and CLP groups were received equal amounts of saline. The serum and liver tissues were collected at 24 h after CLP. The histological changes of the liver were observed by HE staining. The ALT and AST levels in the serum were assessed by using the automatic biochemical analyzer. The levels of cytokines (TNF-α, IL-6, IFN-γ and IL-10) in the serum were quantified by ELISA. MPO activity in the liver tissues were assessed. Western blot was used to detect the expression of pNF-kB p65 and NF-kB p65 in liver tissues. Results Compared with the S group, HE staining in the CLP group showed disordered hepatic cords, hepatocyte swelling and necrosis, infiltrations of inflammatory cells, congestion and bleeding, and the score of liver injury was increased significantly (P<0.05). Levels of ALT, AST, TNF-α, IL-6, IFN-γ, and IL-10 were increased in the CLP group (P<0.05). The activities of NF-κB and MPO in the liver tissues were obviously enhanced (P<0.05). The levels of liver injury, cytokines (TNF-α, IL-6, IFN-γ), MPO and activities of NF-κB in the CLP+PDX group were significantly decreased when compared with those in the CLP group (P<0.05),while the concentration of IL-10 was significantly increased (P<0.05). Conclusions PDX can alleviate sepsis-induced acute liver injury through inhibiting NF-KB activity in the liver tissues.

Objective To explore the safe and effective intravenous anesthetic regimen for intemperants' painless endoscopy.Methods 120 cases of intemperants,aged 25-65 years old,ASA grade Ⅰ or Ⅱ,were randomly divided into 3 groups (n =40):sulfentanyl group (group S),midazolam and sulfentanyl group(group MS),ketamine and sulfentanyl group (group KS).Group S were intravenous injected with sufentanil 0.1 μg/kg,propofol 1-2 mg/kg,etomidate 0.1 mg/kg.Group MS and group KS were additional intravenous injected with midazolam 0.01 mg/kg and ketamine 0.1 mg/kg on the basis of Group S respectively.The occurrence of hypoxia,cough,body movement and blood pressure were recorded,the use of vasoactive drugs and the recovery time were also recorded.Results There are no statistical significant difference of age,gender,and testing time among the three groups (P ＞0.05).Compared with the group S,the total dose of propofol,the incidence of hypoxemia and hypotension,the incidence of body movement and cough reaction in group MS and group KS were all lower (P ＜ 0.01).And compared with the group MS,the patients have lower incidence of hypoxemia and hypotension in group KS (P ＜ 0.05).All the patients were awake well (P ＞ 0.05).Conclusions Small doses of midazolam or ketamine combined with sulfentanyl,propofol and etomidate are safe and effective in the process of anesthesia during endoscopic diagnosis and treatment of intemperants.

Objective To evaluate the reliability of serum catecholamine concentrations in reflecting the depth of dexmedetomidine-based general anesthesia.Methods Forty patients,aged 30-45 yr,of American Society of Anesthesiologists physical status Ⅰ-Ⅱ,undergoing elective laparoscopic hysterectomy under general anesthesia,were divided into 2 groups (n =20 each) using a random number table:dexmedetomidine-based general anesthesia group (group Ⅰ) and general anesthesia group (group Ⅱ).Dexmedetomidine was intravenously infused as a loading bolus of 1 μg/kg over 10 min before induction of anesthesia,followed by an infusion of 0.5 μg · kg-1 · h-1 until 30 min before the end of operation in group Ⅰ.The equal volume of normal saline was given instead of dexmedetomidine in group Ⅱ.Anesthesia was induced with iv midazolam 0.05 mg/kg,propofol 1 mg/kg,cisatracurium 0.15 mg/kg and sufentanil 0.4 μg/kg.Anesthesia was maintained with iv infusion of remifentanil 0.1-0.3 μg · kg-1 · min 1,1％-2％ sevoflurane inhalation and intermittent iv boluses of cisatracurium.Before administration of dexmedetomidine,at the end of administration of the loading dose,at the end of intubation,at the end of skin incision,after establishment of pneumoperitoneum and at the end of operation (T0-5),venous blood samples were taken from the peripheral vein for determination of concentrations of epinephrine (E) and norepinephrine (NE) in serum (by enzyme-linked immunosorbent assay).Results Compared with group Ⅱ],the serum NE and E concentrations were significantly decreased at T1-4 in group Ⅰ (P＜0.05).Compared with the baseline at T0,the serum NE and E concentrations were significantly decreased at T1 in group Ⅰ,and increased at T2-4 in group Ⅱ (P＜0.05).Conclusion The serum catecholamine concentration produces poor reliability in reflecting the depth of dexmedetomidine-based general anesthesia,and thus it is not a suitable monitoring index.

According to Standard Protocol Items: Recommendations for Interventional Trials, Consolidated Standard of Reporting Trials 2010 Statement (CONSORT), CONSORT Extension for Non-Pharmacologic Treatment Interventions (CONSORT for NPT) and Good Clinical Practice, the detailed requirements for protocol writing, reporting, and practicing of clinical trial were classified and summarized in this article. By combining the practical situation of clinical trial of external treatment of TCM such as tuina, the requirernents for clinical trial protocol writing of external treatment of TCM were analyzed and acquired which could improve the quality of clinical trial protocol of external treatment of TCM, thus to provide references for standardized execution of TMC clinical trial and reports of research results.
Clinical Protocols ; standards ; Clinical Trials as Topic ; methods ; standards ; Humans ; Massage ; standards ; Quality Control ; Reference Standards

Objective To integrative the effect of ulinastatin on postoperative cognitive function in elderly gastric cancer surger-y.Methods Two hundred elderly patients with gastric cancer surgery were randomly divided into observed group (100 cases)and control group (100 cases).Patients in observed group received the intravenous drip of ulinastatin before and after the surgery,while others only received the intravenous drip of physiological saline before and after the surgery.Results The urine output of observed group was (441.7±78.5)mL,which was significantly lower than that in control group as the result was (613.2±81.2)mL(P <0.05).After the treatment,the score of MMSE,visual regeneration and association learning in both observed group and control group were significantly lower than that before the treatment(P <0.05).The scores of MMSE,visual regeneration and association learning in observed group were 24.4±1.5,9.7±1.7 and 12.4±1.8,which were significantly higher than that in control group as the scores were 21.1±1.0,8.7±1.5 and 11.3±1.7 (P <0.05).The level of S100βin serum of observed group at the end of sur-gery,1 day and 3 day after the surgery were (0.099±0.024)μg/L,(0.074±0.026)μg/L and (0.061±0.022)μg/L,which were significantly lower than that in control group as the results were (0.138±0.042)μg/L,(0.110±0.034)μg/L and (0.075±0.031)μg/L (P < 0.05).Conclusion Ulinastatin can not only improve the postoperative cognitive dysfunction in elderly patients with postoperative,but can also reduce the level of S100βin serum.It provides brain protection for patients.

Objective To evaluate the cerebral protection of dexmedetomidine during craniotomy under general anesthesia in the patients with craniocerebral injury.Methods Sixty patients with craniocerebral injury,aged 30-50 yr,with body mass index of 18-25 kg/m2,of ASA physical status Ⅱ or Ⅲ,with Glasgow Coma Scale score of 6-12,scheduled for elective craniotomy under general anesthesia,were randomized into 2 groups (n =30 each) using a random number table:control group (group C) and dexmedetomidine group (group Dex).Anesthesia was induced with iv midazolam,propofol,cisatracurium and sufentanil.The patients were endotracheally intubated and mechanically ventilated.In group Dex,dexmedetomidine 1 μg/kg was infused intravenously over 10 min before induction of anesthesia,followed by infusion at a rate of 0.5 μg · kg-1 · h-1 until the end of operation.The equal volume of normal saline was given in group C.Immediately before beginning of surgery (T0),at the moment when the duramater was opened (T1),at 2 h after beginning of surgery (T2),at the duramater closing (T3) and at the end of surgery (T4),blood samples were obtained from the radial artery and jugular venous bulb for blood gas analysis,arteriovenous blood O2 difference and cerebral O2 extraction rate were calculated.The serum concentrations of S-100β were measured by ELISA.Results The serum concentrations of S-100β were significantly increased at T2-4 than at T0 in both groups.The serum concentrations of S-100β were significantly decreased at T2-4 in group Dex than in group C.The parameters of cerebral oxygen metabolism were all within the normal range in both groups.Conclusion Dexmedetomidine (1 μg/kg infused intravenously before induction of anesthesia,followed by infusion at a rate of 0.5 μg · kg-1 · h-1 until the end of operation) provides cerebral protection to some extent during craniotomy under general anesthesia in the patients with craniocerebral injury.