ObjectiveThis study aims to investigate the effect of Dictamni Cortex on intestinal barrier damage in rats and its mechanism by untargeted metabolomics and targeted metabolomics for short-chain fatty acids （SCFAs）. MethodsRats were randomly divided into a control group， a high-dose group of Dictamni Cortex （8.1 g·kg^-1）， a medium-dose group （2.7 g·kg^-1）， and a low-dose group （0.9 g·kg^-1）. Except for the control group， the other groups were administered different doses of Dictamni Cortex by gavage for eight consecutive weeks. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in the ileal tissue. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the level of cytokines， including tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β）， in the ileal tissue of rats. Quantitative real-time fluorescence polymerase chain reaction （Real-time PCR） technology was used to detect the expression level of tight junction proteins， including zonula occludens-1 （ZO-1）， Occludin， and Claudin-1 mRNAs， in the ileal tissue of rats to preliminarily explore the effects of Dictamni Cortex on intestinal damage. The dose with the most significant toxic phenotype was selected to further reveal the effects of Dictamni Cortex on the metabolic profile of ileal tissue in rats by non-targeted metabolomics combined with targeted metabolomics for SCFAs. ResultsCompared with the control group， all doses of Dictamni Cortex induced varying degrees of pathological damage in the ileum， increased TNF-α （P<0.01）， IL-6 （P<0.01）， and IL-1β （P<0.01） levels in the ileal tissue， and decreased the expression level of ZO-1 （P<0.05， P<0.01）， Occludin （P<0.01）， and Claudin-1 （P<0.05） in the ileal tissue， with the high-dose group showing the most significant toxic phenotypes. The damage mechanisms of the high-dose group of Dictamni Cortex on the ileal tissue were further explored by integrating non-targeted metabolomics and targeted metabolomics for SCFAs. The non-targeted metabolomics results showed that 21 differential metabolites were identified in the control group and the high-dose group. Compared with that in the control group， after Dictamni Cortex intervention， the level of 14 metabolites was significantly increased （P<0.05， P<0.01）， and the level of seven metabolites was significantly decreased （P<0.05， P<0.01） in the ileal contents. These metabolites collectively acted on 10 related metabolic pathways， including glycerophospholipids and primary bile acid biosynthesis. The quantitative data of targeted metabolomics for SCFAs showed that Dictamni Cortex intervention disrupted the level of propionic acid， butyric acid， acetic acid， caproic acid， isobutyric acid， isovaleric acid， valeric acid， and isocaproic acid in the ileal contents of rats. Compared with those in the control group， the level of isobutyric acid， isovaleric acid， and valeric acid were significantly increased， while the level of propionic acid， butyric acid， and acetic acid were significantly decreased in the ileal contents of rats after Dictamni Cortex intervention （P<0.05， P<0.01）. ConclusionDictamni Cortex can induce intestinal damage by regulating glycerophospholipid metabolism， primary bile acid biosynthesis， and metabolic pathways for SCFAs.

Kisspeptin, the neuropeptide produced by Kiss1 neurons in the hypothalamus, is involved in the neuroendocrine regulation of puberty initiation, reproductive system maturation, ovulation and other processes by influencing the secretion of gonadotropin-releasing hormone. Kiss1 gene expression is regulated by multiple trans-regulatory factors and epigenetics. Prediction and preliminary experiments have shown that the seed sequences of miR-92a-3p and miR-25-3p can directly bind to the 3′-UTR of Kiss1 and inhibit the expression of Kiss1. In order to further study the role of miR-92a-3p and miR-25-3p in the regulation of Kiss1, specific absorptive sponge vectors (sponge-miR-92a and sponge-miR-25) with inhibitory effects on miR-92a-3p and miR-25-3p were constructed to realize the functional loss of miRNA. Flow cytometry and dual luciferase reporter assays both confirmed that both sponge vectors could adsorb exogenous or endogenous target miRNAs very effectively. The sponge-miR-92a and sponge-miR-25 vectors are further packaged into the lentivirus LV-sponge-miR-92a and LV-sponge-miR-25. The results of real-time fluorescence quantitative PCR showed that the expression level of Kiss1 in the hypothalamic primary neurons infected by LV-sponge-miR-92a and LV-sponge-miR-25 was significantly up-regulated (P < 0. 05). After injecting LV-sponge-miR-92a into the hypothalamus, the time of female mouse vulva opening was significantly earlier (P<0. 05). The normal oestrus cycle of female mice with was disrupted by injections of LV-sponge-miR-92a and LV-sponge-miR-25 in the hypothalamus. In conclusion, we successfully constructed sponge vectors capable of effectively adsorbing miR-92a-3p and miR-25-3p, and demonstrated their role in removing the inhibition of miR-92a-3p and miR-25-3p on Kiss1. Hypothalamic sponge injection had a certain effect on both the time of vulva opening and the estrus cycle of female mice, suggesting that miR-92a-3p and miR-25-3p may play an important role in the initiation of puberty and reproductive maturity.

Objective: To investigate the prevalence, mutation characteristics and clinical outcomes of inherited metabolic diseases(IMD) by using tandem mass spectrometry screening. Methods: In Hunan province, 565 182 newborns who underwent tandem mass spectrometry (MS/MS) screening for IMDs were studied, including fatty acid oxidation disorders (FAODs), amino acid disorders (AAs), and organic acidemias (OAs) between March 2013 and September 2017.For the patients with positive results, a recall screening test was performed, and the results were further confirmed by specific biochemical and genetic analysis.For all the patients with IMD, guideline-directed medical treatment was administrated, and the follow-up outcomes was evaluated. Results: A total of 107 newborns were diagnosed with IMDs, with an overall prevalence of 1∶5 282, including 65 newborns with FAODs (1∶ 8 695), 29 newborns with AAs (1∶19 489), and 13 newborns with OAs (1∶43 476). The primary carnitine deficiency(PCD)(44 cases), hyperphenylalaninemia (HPA)(17 cases), short-chain acyl-CoA dehydrogenase deficiency(SCADD)(12 cases), citrine deficiency(NICCD)(6 cases) were the 4 most common IMDs in Hunan province.The hotspot mutations in SLC22A5 gene of PCD were c. 51C>G(25.3%), c.1400C>G(23.0%), and c. 760C>T(13.8%); in PAH gene of HPA were c. 728G>A (22.2%) and c. 721C>T(14.8%); in ACADS gene of SCADD was c. 1031A>G(38.9%); and in SLC25A13 gene of NICCD was c. 851_854delGTAT (50.0%), respectively.The remaining IMDs were rare, and the hotspot mutations were unclear right now.During a mean follow-up of (26.1±5.6) months, 7 patients died, 4 patients suffered an intelligent disability, whereas the remaining 96 subjects had normal physical and intelligent development. Conclusions The overall prevalence of IMDs is not fairly low in Hunan province.Newborn screening and early appropriate management can significantly improve the outcomes of these patients.

Objective To investigate the effects of thyroid hormone on extremely severe traumatic brain injury (TBI).Methods A retrospective case-control study was conducted to analyze the treatment of 105 patients with extremely severe TBI admitted from July 2010 to April 2014.There were 79 males and 26 females,with an average age of 32.9 years.The patients were divided into conventional treatment group (Group A,35 cases),conventional treatment ± thyroxine treatment group (Group B,35 cases) and thyroxine treatment group after the condition that thyroxine level was low (Group C,35 cases) according to the random number table method.The incidence of low T3 and T4,incidence of hypotension,the dosage of vasoactive drugs,function evaluation of liver and kidney damage,Glasgow outcome scale (GOS),and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) within 20 days after admission,and mortality rate within 30 days after admission were compared and analyzed.Results Within 20 days after admission,the rates of low thyroxine levels and hypotension of the Group B (22.9％,77.1％) were significantly lower than those in the other two groups (Group A:40％,100％;Group C:37％,100％) (all P ＜ 0.05).The doses of dopamine and norepinephrine in Group B was significantly lower than the other two groups and the combination rate of vasopressors in Group B was significantly lower than the other two groups (P ＜ 0.05),while there was no significant difference between Group A and Group C (P ＞ 0.05).The corresponding data in Group A and Group C had no statistically significant difference (P ＞ 0.05).The liver and renal dysfunction rates of Group B (29％,31％) were significantly lower than those of the other two groups (Group A:49％,51％;Group C:43％,51％) (all P ＜ 0.05).The corresponding data in Group A and Group C had no statistically significant difference (P ＞ 0.05).GOS in Group B [(4.8 ± 1.9) points] was significantly higher than that in Group A [(3.3 ± 0.2) points] (all P ＜ 0.05) within 30 days after admission and significantly higher than that of itself at the beginning [(3.6 ± 1.1) points] (P ＜ 0.05).The APACHE Ⅱ in Group A was significantly higher than those in other two groups as well as that in Group A at admission (P ＜ 0.05).Mortality rates in Group B (31％) and Group C (29％) were significantly lower than that in Group A (69％) within 30 days after admission (P ＜ 0.05).Conclusions Thyroxine can reduce the incidence of hypotension,liver and kidney injury rate in extremely severe TBI.Prevention is better than the supplementary treatment after severe TBI.Thyroxine can also reduce the mortality of extremely severe TBI within 30 days after admission.

Objective To discuss the clinical characteristics and prognosis of lipoprotein glomerulopathy (LPG) in chil-dren. Method Clinical data of one pediatric LPG patient were retrospectively analyzed. The clinical features and prognosis of childhood LPG were summarized based on literature review. Results A nine years old girl presented with frequent urination. The ifrst urine test revealed hematuria and proteinuria. After one week anti-infection treatment, the hematuria and proteinuria were continued. The serum albumin was slightly reduced. The hyperlipidemia and mild anemia were emerged. Kidney biopsy showed that enlarged glomeruli, with dilated capillary loops and weak eosinophilic lipoprotein thrombi in the capillary lumina under the light microscope;layered or tuftedemboluscontaining particulated lipid vacuoles under electron microscope. Gene sequencing identified APOE Tokyo (Leu141-Lys143→0). The diagnosis of LPG was confirmed. The lipid-lowering therapy was administrated and the disease was alleviated. Conclusion LPG is a rare disease in children. The level of blood lipid was signiifcantly increased, and the hormone therapy was ineffective. Kidney biopsy is the main basis for diagnosis. The genetic testing can prompt the genetic background. Lipid lowering therapy can relieve the progress of the disease.

Objective To evaluate the occupational competency of clinical post-graduate students by the 360-degree feedback system.Methods The occupational competencies of the 102 clinical post-graduate students were evaluated by department directors, teachers, the superior band taught physicians, peers, post-graduate themselves using a self-designed questionnaire.The evaluation content of professional competence included 12 core elements, such as medical knowledge, clinical skills, and professional ethics and so on.EpiData 3.02 was used to establish the database, SPSS 19.0 software was used to carry on the statistical analysis, and the statistical analysis method was mainly for the descriptive statistics analysis, the single factor analysis of variance.Results 5 kinds of evaluation subjects were statistically significant (P＜0.05) in medical knowledge, clinical skills, clinical thinking ability, lifelong learning ability, professional ethics, team cooperation ability, self cognition and management ability and total score.Students themselves and their peers scored high in various aspects, teaching superior doctors rated middle and the director and mentor gave low grade in all aspects and the scores rated by the department directors and teachers were lower than students themselves(P=0.003, P=0.047).Conclusion The 360-degree evaluation system can comprehensively and objectively evaluate the clinical post-graduate students' occupational competency and offer beneficial assistant to clinic and practice.

Objectives It was constructed that the replication defective adenoviral vectors carried the short hairpin sequences of mouse SCP2.And we will make a further study of mechanism between SCP2 gene and cholesterol stone in gallbladder.Methods The short hairpin sequences of mouse SCP2 were cloned by two-step PCR,and connected together with the plasmid pDC312.The Admax Adenoviral Vector System was used to generate the replication defective adenoviral vectors,which were purified by CsCl method.The processes of TCID50 were applied to detect the titers of the adenoviral vectors.Furthermore,Protein levels of SCP2 were determined by Western blot analysis,and the levels of SCP2、CYP7A1、HMGCR mRNA from the hepa1-6 cell of mouse were measured by the usage of RT-PCR.Results SCP2mRNA and SCP2 protein were down-regulated by the replication defective adenoviral vectors carried the SCP2-shRNA.With the decreasing SCP2mRNA,the levels of HMGCRmRNA were down-regulated at same the time,while CYP7A1mRNA were up-regulated.Conclusions The replication defective adenoviral vectors carried SCP2-shRNA were constructed successfully.The lower levels of SCP2 could affect the activities of HMG-CoA reductase and CYP7-a enzyme,which caused the variations of cholesterol metabolism and then decreased the formation of cholesterol stone.

Comprehensively improving the quality of postgraduate education is the main theme of reform and development of postgraduate education. A series of effective measures was taken in the First Affiliated Hospital of Harbin Medical University according to the various aspects of postgraduate education, implementing the whole quality control, enhancing the quality of postgraduate education, in order to accommodate the new situation of postgraduate education.

Scientific and rigorous management system is the prerequisite for specialists to standardize training.In order to cultivate and promote the actual clinical ability,the First Clinical College of Harbin Medical University establish a scientific,reasonable,standard and strict evaluation system in specialist training - the Mini-CEX.This system was employed to test on completion of a course and meet the current medical development of specialist training and evaluation mode.