BACKGROUND:Sepsis has become the greatest threat to in-patients, with a mortality of over 25％.The dysfunction of gut barrier, especially the immunological barrier, plays an important role in the development of sepsis. This dysfunction occurs after surgery, but the magnitude of change does not differentiate patients with sepsis from those without sepsis. Increased intestinal permeability before surgery is of no value in predicating sepsis. The present study aimed to observe the changes of intestinal mucosal immunologic barrier in rat models of sepsis induced by cecal ligation and puncture. METHODS:Sixty Sprague-Dawley rats were randomly divided into a sepsis group (n=45) and a control group (n=15). The rats in the sepsis group were subjected to cecal ligation and puncture (CLP), whereas the rats in the control group underwent a sham operation. The ileac mucosa and segments were harvested 3, 6 and 12 hours after CLP, and blood samples were collected. Pathological changes, protein levels of defensin-5 (RD-5) and trefoil factor-3 (TFF3) mRNA, and lymphocytes apoptosis in the intestinal mucosa were determined. In an additional experiment, the gut-origin bacterial DNA in blood was detected. RESULTS:The intestinal mucosa showed marked injury with loss of ileal villi, desquamation of epithelium, detachment of lamina propria, hemorrhage and ulceration in the sepsis group. The expression of TFF3 mRNA and level of RD-5 protein were decreased and the apoptosis of mucosal lymphocyte increased (P<0.05) in the sepsis group compared with the control group. Significant differences were observed in RD-5 and TFF3 mRNA 3 hours after CLP and they were progressively increased 6 and 12 hours after CLP in the sepsis group compared with the control group (P<0.05, RD-5 F=11.76, TFF3 F=16.86 and apoptosis F=122.52). In addition, the gut-origin bacterial DNA detected in plasma was positive in the sepsis group. CONCLUSION:The immunological function of the intestinal mucosa was impaired in septic rats and further deteriorated in the course of sepsis.

OBJECTIVETo understand the relationship between the HIV-1 viral sequence variation and host factors associated with HIV-1 disease progression.

METHODSEnv and gag fragments of HIV-1 were amplified with PCR, cloned and sequenced. Bioinformatics was employed to find the genetic variation, N-linked glycosylation, hypermutation etc. Host gene polymorphism was analysed by using restricted fragment length polymorphism (RFLP).

RESULTSSignificant difference was found in genetic divergence between Env PCR dominant and clonal sequences (0.1 and 0.06, respectively) in non-treated group, but no significant difference was found in the HAART treated group. V3 GPGQ accounted for the most part in both treated and nontreated groups, rare V3 loop such as GPGH, GQGR and GLGR was found in treated group, V3 substitutions of I/V (position 12) and Y/H (position 21) was associated with the relatively rapid progression (RRP). Glycosylation was significantly higher in RRP than in TP for Env region, GA substitution in RRP was also significantly higher than that in TP group. SDF1-3primeA and CCR2 V64I gene frequency was higher in TP than in RRP, but the difference was not significant.

CONCLUSIONDisease progression was associated with V3 AA change, glycosylation and GA substitution in env gene. SDF1-3primeA, CCR2 V64I and CX3CR1 V249I/M280T was not associated with disease progression significantly.

Adult ; Disease Progression ; Female ; Genetic Variation ; Glycosylation ; HIV Infections ; pathology ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; metabolism ; Humans ; Male ; Phylogeny ; Polymorphism, Genetic ; Receptors, Chemokine ; genetics ; env Gene Products, Human Immunodeficiency Virus ; genetics ; metabolism ; gag Gene Products, Human Immunodeficiency Virus ; genetics ; metabolism

Objective To observe the effects of estradiol benzoate (E_2),human cberionic gonadotropin (hCG) and luteinizing hormone-releasing hormone (LHRH) on testicular descent and their relation to the expression of insulin-like factor-3 (INSL3) mBNA in rats.Methods At birth,40 SD male rats were assigned to 4 groups,and each group consisted of 10 rats.Each group received injection of one of the following drugs:normal saline(NS),E_3,hCG+E_2,and LHRH+E_2 for 29 clays after birth.Total RNA was extracted from the testis on day 30 after birth.INSL3 was synthesized and further amplified by RT-PCR.Results (1) Testicular descent rate on clay 30 after birth was 100% (20/20) in the group of NS,0% (0/20) in the group of E_2,85% (17/20) of hCG + E_2,and 70% (14/20) of LHBH + E_2.There was no significant difference between the last two groups ( P = 0.45).(2) Weight of testis differed significantly among the groups ( F = 56.67,P＜0.01 ),being lightest in the group of E_2.(3) E_2 could damage the germ cells of testis,however hCG and LHRH ameliorated this situation. (4) The expression of INSL3 mBNA was absent in the group of E_2,while the other groups showed normal expression.Conclusion E_2 inhibits testicular descent in rats,while hCG and LHRH promote the progress.The mechanism seems to be related to the expression of INSL3 mRNA.

OBJECTIVETo identify the factors that affect the safety and efficacy of peroral endoscopic myotomy (POEM) for treatment of achalasia.

METHODSData of consecutive patients undergoing POEM for confirmed achalasia between December, 2010 and December, 2015 were collected, including the procedure time, approach of tunnel entry incision, approach of myotomy, complications and follow-up data.

RESULTSAmong the total of 439 patients enrolled, the overall complication rate was 28.7% (126/439). Treatment success (Eckardt score≤3) was achieved in 94.5% of 364 patients followed up for a median of 6 months (1-48 months), and the mean score was reduced significantly from 6.7∓1.5 before treatment to 1.2∓1.1 after the treatment (P<0.05). Logistic regression revealed that the year when POEM was performed and the approach of entry incision were two significant factors contributing to complications: with the year 2015 as the reference, the odds ratio (OR) was 9.454 (95% CI: 2.499-35.76) for the years before 2011, 2.177 (95% CI: 0.794-5.974) for 2012, 3.975 (95% CI: 1.904-8.298) for 2013, and 1.079 (95% CI: 0.601-1.940) for 2014; with the longitudinal entry incision as the reference, the OR was 0.369 (95% CI: 0.165-0.824) for inverted T entry incision and 0.456 (95% CI: 0.242-0.859) for transverse entry incision. The approach of myotomy was the significantly associated with symptomatic relapse: with full-thickness myotomy combined with indwelling an anti-reflux belt as the reference, the OR was 0.363 (95% CI: 0.059-2.250) for gradual full-thickness myotomy, 2.137 (95% CI: 0.440-10.378) for circular muscle myotomy, and 4.385 (95% CI: 0.820-23.438) for circular muscle myotomy in combination with balloon shaping; the recurrence rate was 0 with a full-thickness myotomy.

CONCLUSIONThe complication rates of POEM appears to decrease over time, and an inverted T entry incision is the best choice for controlling the complications. Gradual full-thickness myotomy is an excellent approach for treatment of achalasia in terms of the relapse rate, procedure time and the incidence of reflux esophagitis.

Endoscopy ; Esophageal Achalasia ; surgery ; Esophagitis, Peptic ; surgery ; Gastroesophageal Reflux ; Humans ; Muscles ; surgery ; Recurrence ; Treatment Outcome

Objective:To investigate the curative efficacy of modified Qilang prescription on drug-dependent constipation with Qi and Yin deficiency and the effects on serum vasoactive intestinal peptide （VIP）， motilin （MTL）， 5-hydroxytryptamine （5-HT）， and 5-hydroxytryptamine 4 receptor （5-HT4R）. Method:A total of 160 patients diagnosed with drug-dependent constipation were randomly divided into a treatment group （n=80， Qilang prescription） and a control group （n=80， lactulose oral solution）. The treatment lasted for eight weeks. Changes in clinical symptoms， traditional Chinese medicine （TCM） syndrome， and serum VIP， MTL， 5-HT， and 5-HT4R before and after treatment were observed. The clinical efficacies of the two groups were compared. An eight-week follow-up was carried out for the observation of recurrent rate and TCM syndrome. Result:The overall response rate of the treatment group （90.91%） was higher than that （75.00%） of the control group （Z=-6.514，P<0.05）. There was no significant difference in serum VIP， MTL， 5-HT， and 5-HT4R between the two groups before treatment. After treatment for eight weeks， both groups showed reduced serum VIP level as compared with those before treatment， and the treatment group was inferior to the control group （P<0.05）. The serum MTL levels of the two groups were both higher than those before treatment （P<0.05）， and the treatment group was superior to the control group （P<0.05）. After treatment， the level of 5-HT in the treatment group was higher than that in the control group （P<0.05）. The post-treatment 5-HT4R level in the treatment group slightly increased （P<0.05）， but no significant difference in 5-HT4R levels between the two groups after treatment was observed. During the eight-week follow-up， the recurrence rate in the treatment group was significantly lower than that in the control group at the 2nd and 4th weeks （P<0.05）. There was no significant difference in the recurrence rate between the treatment group ［57.14% （40/70）］ and the control group ［64.81% （35/54）］ after eight weeks. Conclusion:Modified Qilang prescription was superior to lactulose in the short- and mid-term efficacy on drug-dependent constipation with Qi and Yin deficiency. No significant difference in the long-term efficacy was observed. The underlying therapeutic mechanism might be related to the regulation of serum VIP， MTL， 5-HT， and 5-HT4R levels.

Objective: To investigate the effect and molecular mechanism of ultra-conservative long non-coding RNA uc.77 in lung cancer. Methods: Lung cancer tissues and adjacent normal tissues were obtained from 61 patients with lung cancer who were diagnosed with lung cancer and underwent surgery from 2014 to 2016 in the General Hospital of the Southern Theater Command of the People's Liberation Army. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the uc.77 relative expressions in normal human bronchial epithelial cells 16HBE, lung cancer cell lines, and 61 pair lung cancer tissues. Uc.77 siRNA was transfected into lung cancer cells to interfere with the expression of uc.77, qRT-PCR was used to verify the interference effect, CCK8 method and clone formation experiment were used to detect cell proliferation ability, flow cytometry was used to detect apoptosis and cell cycle changes. H1299 cells transfected with uc.77 siRNA were injected into the subcutaneous right side of BALB/c nude mice to construct a tumor-bearing model for exploring the role of uc.77 on tumor growth. Western blot and qRT-PCR methods were used to detect the protein and mRNA expressions of p21. Results: The relative expression levels of uc.77 in lung cancer cell lines 95D, H1299, A549, H460, H446 and 16HBE-T were significantly higher than that of 16HBE cells (P<0.05). The uc.77 RNA expression levels of lung cancer tissues was significantly higher than that of the adjacent normal tissues (P<0.001). In addition, increased lncRNA uc.77 expression was significantly associated with big tumor size, lymph node metastasis and advanced TNM stage (P<0.05). After transfection with uc.77 siRNA, the expressions of uc.77 in H1299, 95-D and 16HBE-T cells were reduced (P<0.05), and the cell proliferation capacities were reduced at 48 hours and 72 hours (P<0.05). After transfection with uc.77 siRNA-1, the G(0)/G(1) phase cell ratio of H1299 siRNA-1 group [(71.86±3.46)%] was higher than those of H1299-control group [(47.62±5.48)%] and H1299 siRNA-NC group [(61.38±5.62)%, P<0.05], S phase cell ratio of H1299 siRNA-1 group [(14.99±3.61)%] was lower than those of H1299-control group [(34.95±7.05)%] and H1299 siRNA-NC group [(23.75±5.87)%, P<0.05], the apoptosis rate of H1299 siRNA-1 group [(4.90±1.80)%] was higher than those of H1299-control group [(3.30±0.80)%] and H1299 siRNA-NC group [(2.80±1.20)%, P<0.05], the colony formation rate of H1299 siRNA-1 group [(19.20±2.00)%] was lower than those of H1299 control group [(32.60±2.00)%] and H1299 siRNA-NC group [(34.40±1.00)%, P<0.05]. The results of the nude mice tumor formation experiment showed that the tumor volume of the H1299 siRNA-1 group was significantly lower than those of the H1299-control group and the H1299-negative control group (P<0.05), the average tumor weight of H1299 siRNA-1 group was significantly lower than those of H1299-control group and H1299-negative control group (P<0.05), tumor cell growth marker Ki-67 in the H1299 siRNA-1 group showed weak positive, and Ki-67 in the H1299-control group and H1299-negative control group showed positive. The result of qRT-PCR analysis showed that the mRNA expression level of p21 in H1299 siRNA-1 group (2.57±0.45) was higher than those in H1299 control group (1.00±0.00, P=0.001) and H1299 siRNA-NC group (1.52±0.37, P=0.009). The result of western blotting analysis also showed that the expression of p21 protein level in H1299 siRNA-1 group increased. Conclusions: The expression of ultraconserved long non-coding RNA uc.77 is elevated in lung cancer cell lines and lung cancer tissues. Silencing the expression of ultraconservative long noncoding RNA uc.77 can inhibit tumor growth, and blocking uc.77 expression may be a potential therapeutic target for lung cancer.
Mice ; Animals ; Humans ; RNA, Long Noncoding/metabolism* ; Mice, Nude ; RNA, Small Interfering/metabolism* ; Ki-67 Antigen/metabolism* ; Cell Line, Tumor ; Lung Neoplasms/pathology* ; Cell Proliferation ; Apoptosis/genetics* ; RNA, Messenger ; Gene Expression Regulation, Neoplastic

Objective: To investigate the effectiveness of nucleos(t)ide analogues in the treatment of HBeAg-positive chronic hepatitis B with normal alanine aminotransferase and high level of HBV DNA. Methods: Treatment-naïve chronic hepatitis B patients who were followed up at the Center of Infectious Diseases, West China Hospital of Sichuan University from January 2019 to January 2020 were selected as subjects. Demographic characteristics, the results of laboratory examination before treatment and one year after treatment were retrospectively collected. Patients were divided into tenofovir dipivoxil (TDF) and propofol fumurate tenofovir (TAF) treatment group according to different types of medication. The changes of serum HBV DNA level, HBeAg serological conversion and HBsAg quantitative level were analyzed and compared between the two groups. Results: A total of 38 cases were enrolled. Among them, there were 16 and 22 cases in the TDF and TAF group, respectively. There was no statistically significant difference in demographic characteristics, baseline HBV DNA levels and HBsAg quantitative levels between the two groups. Virological response was achieved in 60.5% (23/38) of patients after one year of antiviral therapy. Serum HBV DNA levels below the lower limit of detection [68.2% (15/22) vs. 50.0% (8/16), P=0.258] and higher HBeAg seroconversion rate [18.2%] (4/22) vs. 6.3% (1/16), P=0.374] was obtained in TAF than TDF group; however, there was no statistically significant differences between the two. Serum HBsAg quantitative level was significantly reduced with TDF and TAF treatment. In addition, alanine aminotransferase elevation was reduced in TAF than TDF treated group. Multivariate logistic regression analysis showed that patient age was an independent predictor of a virological response to antiviral therapy. Conclusion: HBeAg-positive CHB patients with normal alanine aminotransferase, and high HBV DNA level can obtain better curative effect after TDF and TAF treatment.
Alanine Transaminase ; Antiviral Agents/therapeutic use* ; DNA, Viral ; Hepatitis B Surface Antigens ; Hepatitis B e Antigens ; Hepatitis B virus/genetics* ; Hepatitis B, Chronic ; Humans ; Retrospective Studies ; Tenofovir/therapeutic use* ; Treatment Outcome

Tumor is one of the diseases that seriously endanger human health， and how to treat tumor effectively is still one of the important problems in the field of medicine. At present， most of the radiotherapies and chemical drugs for cancer have serious side effect despite of an obvious efficacy. With a unique syndrome differentiation treatment system and overall concept， traditional Chinese medicine has become the key research and development object of antitumor drugs due to many advantages， such as multiple channels， multiple levels， multiple links， multiple targets and less toxicity， and could can fully mobilize the immune and epidemic prevention mechanism of the body. A large number of studies have shown that Xiao Xianxiongtang and its effective ingredients have obvious antitumor effect. Many doctors have applied Xiao Xianxiongtang and modified formulas in clinical treatment of tumors， and relevant pharmacological studies have also confirmed the effectiveness of this formula， but with a lack of systematic summary of its effective ingredients and its mechanism of action. Now， with alkaloids， ketones， sterols and phenols in Xiao Xianxiongtang as the starting point， this study mainly focuses on inhibition of tumor cell proliferation， invasion and migration， induction of tumor cell apoptosis and autophagy， inhibition of tumor cell cycle， enhancement of tumor cell sensitivity， inhibition of tumor angiogenesis and regulation of immunosuppressive tumor microenvironment from two ways to sort out composition， function and mechanism of drugs. In this paper， effective components， main targets and mechanism of intervention in the tumor development of Xiao Xianxiongtang were reviewed， in order to provide a new idea for subsequent antitumor research and development of this prescription.

OBJECTIVE: To analyze the cement flow in the abutment margin-crown platform switching structure by using the three-dimensional finite element analysis, in order to prove that whether the abutment margin-crown platform switching structure can reduce the inflow depth of cement in the implantation adhesive retention. METHODS: By using ANSYS 19.0 software, two models were created, including the one with regular margin and crown (Model one, the traditional group), and the other one with abutment margin-crown platform switching structure (Model two, the platform switching group). Both abutments of the two models were wrapped by gingiva, and the depth of the abutment margins was 1.5 mm submucosal. Two-way fluid structure coupling calculations were produced in two models by using ANSYS 19.0 software. In the two models, the same amount of cement were put between the inner side of the crowns and the abutments. The process of cementing the crown to the abutment was simulated when the crown was 0.6 mm above the abutment. The crown was falling at a constant speed in the whole process spending 0.1 s. Then we observed the cement flow outside the crowns at the time of 0.025 s, 0.05 s, 0.075 s, 0.1 s, and measured the depth of cement over the margins at the time of 0.1 s. RESULTS: At the time of 0 s, 0.025 s, 0.05 s, the cements in the two models were all above the abutment margins. At the time of 0.075 s, in Model one, the gingiva was squeezed by the cement and became deformed, and then a gap was formed between the gingiva and the abutment into which the cement started to flow. In Model two, because of the narrow neck of the crown, the cement flowed out from the gingival as it was pressed by the upward counterforce from the gingival and the abutment margin. At the time of 0.1 s, in Model one, the cement continued to flow deep inside with the gravity force and pressure, and the depth of the cement over the margin was 1 mm. In Model two, the cement continued to flow out from the gingival at the time of 0.075 s, and the depth of the cement over the margin was 0 mm. CONCLUSION When the abutment was wrapped by the gingiva, the inflow depth of cement in the implantation adhesive retention can be reduced in the abutment margin-crown platform switching structure.
Finite Element Analysis ; Cementation/methods* ; Gingiva ; Crowns ; Dental Abutments ; Dental Cements ; Dental Stress Analysis

Next-generation sequencing has revolutionized family-based association tests for rare variants. As the lower power of genome wide association study for detecting casual rare variants, methods aggregating effects of multiple variants have been proposed, such as burden tests and variance component tests. This paper summarizes the methods of rare variants association test that can be applied for family data, introduces their principles, characteristics and applicable conditions and discusses the shortcomings and the improvement of the present methods.
Computer Simulation ; Family Relations ; Genetic Association Studies ; Genetic Variation ; Genome-Wide Association Study/methods* ; Humans