Objective To investigate the role of p53 in the regulation of heat shock protein(Hsp)84 and 86,and the correlation of their functional imbalances with accelerated brain aging and with suppressed tumorigenesis in SAMP8 mice(senescence accelerated mouse prone 8).Methods The mRNA and protein expressions of Hsp84 and Hsp86,and protein expressions of p53 pathway-related proteins(p21 and MDM2)in hippocampus of SAMP8 mice and their control SAMR1(senescence accelerated mouse resistant 1)mice were determined.Murine Neuro-2a cells were treated with 20 μmol/L Aβ25-35,and then mRNA expressions of p53,Hsp84 and Hsp86 in these cells were detected.Neuro-2a cells were co-transfected with p53 siRNA and pHsp84-Luc or pHsp86-Luc plasmid and treated with 20 μmol/L Aβ25-35,then promoter activity of Hsp84 and Hsp86 were detected in these cells.After co-transfection with pcDNA3.1-p53 or pcDNA3.1-p53DD and pHsp84-Luc or pHsp86-Luc plasmids,the neuro-2a cells were treated with 20 μmol/L Aβ25-35.Then promoter activity of Hsp84 and Hsp86 were detected in these cells at different concentrations of p53.Results The mRNA levels of Hsp84 and Hsp86 in the hippocampus of SAMP8 mice were significantly declined,which were 13.51% and 16.13% of SAMR1 mice,respectively(all P<0.01).Compared with the SAMR1 mice,the protein expressions of Hsp84 and Hsp86 in the hippocampus of SAMP8 mice were obviously declined(all P<0.01).Whereas,p53 pathway-related protein p21 expression was increased and MDM2 expression was decreased(all P<0.01).The mRNA expression of p53 in AD cells was significantly increased by 58%(P<0.01),whereas Hsp84 and Hsp86 mRNA levels were significantly decreased by 32% and 41%,respectively as compared with the normal cells(all P<0.05).Inhibition of p53 in AD cells could increase promoter activity of Hsp84 and Hsp86 significantly in a concentration-dependent manner(both P<0.05),whereas overexpression of p53 in the cells could lead to decreased promoter activity of them in a concentration-dependent manner(both P<0.05).Conclusions The p53 can negatively regulate the expressions of Hsp84 and Hsp86.The activity of p53/p21 pathway is increased,while Hsp84 and Hsp86 are inhibited in the brain of SAMP8 mice.Functional imbalance between p53 and Hsp84/86 might be the part of reasons causing accelerated aging and suppressed tumorigenesis in SAMP8 mice.

AIM: To investigate the biocolonization of poly 2-hydroxyethy methacrylate (PHEMA) sponge with cornea tissue and evaluate the therapeutic effects of modified porous poly 2-hydroxyethy methacrylate-Polymethyl met-hacrylate (PHEMA-PMMA) Keratoprostheses (KPro) on rabbit and monkey corneas. METHODS:The KPro were made using two-stage polymerization combined with mechanical cutting. The experiment was divided into two groups. In the control group, ten normal rabbit eyes received lamellar implanta-tion of PHEMA sponges. The sponges were obtained 2 weeks, 1,2,3 and 4 months after operation. The cell proliferation and neovascularization inside the sponges were observed using light and transmission electron microscopy (TEM) and immunohistochemistry. In the experimental group, the porous PHEMA-PMMA KPros were inserted into the lamellar pockets of eight rabbit corneas and two monkey corneas (stage I operation). The healing process was investigated by slit-lamp microscopy. The anterior lamellar cornea tissues were removed 3 months after surgery, exposing the under-neath transparent core (stage II operation). The operated eyes were then followed up for 3-6 months.light microscope, fibroblasts started to grow into the cornea 2 weeks after operation; lots of cells, accompanied with new blood vessels, invaded into the cornea 2-3 months after surgery. Invading cells of sponge, as well as keratocytes, were positive for vimentin. Under the electron microscope, the invading cells looked healthy and were surrounded by extracellular matrix and collagen. In 8 rabbit eyes which received KPro implantation, anterior lamellar cornea melting happened in two eyes after the stage II operation. The remaining 6 corneas retained their central cores during observation after the stage II operation.Two monkey operated eyes were found no complication thoughout the whole follow-up.cornea. The modified PHEMA-PMMA KPros have obtained a relatively stable results after implantation into animal corneas.

Bombyx mori nucleopolyhedrovirus (BmNPV) bm47 gene is found in all sequenced lepidopteran nucleopolyhedroviruses (NPVs). It is one of the core genes of NPVs. However, the role of bm47 in the biological cycle of NPV remains unknown. In this study, the Red recombination system was used to knock out bm47 from BmNPV to construct bm47-ko-Bacmid in E. coli BW25113 system. Then bm47 gene was introduced back to the viral genome using the Bac-to-Bac system to create the repair virus bm47-re-Bacmid. TCID50 assay and real-time PCR (qPCR) were used to evaluate the effects of bm47 deletion on viral DNA replication, gene transcription, and protein expression. qPCR results showed that bm47 knock-out had no significant effect on viral DNA replication. However, the qPCR results showed that bm47-ko-Bacmid significantly decreased the transcription levels of early gene lef-3, late gene vp39, and very late gene p10 at 48 h and 72 h after viral transfection of BmN cells (P < 0.05). This work will provide a foundation for further studies on the biological function of BmNPV bm47 in viral replication and transcription.
Animals ; Bombyx ; virology ; Gene Deletion ; Gene Expression Regulation, Viral ; Nucleopolyhedrovirus ; genetics ; physiology ; Transcription, Genetic ; Viral Proteins ; genetics ; metabolism ; Virus Replication

OBJECTIVETo assess the effect of intra-abdominal hypertension (IAH) on hemodynamics of severe acute pancreatitis (SAP) in porcine model.

METHODSFollowing baseline registrations, SAP was induced in 12 animals. The N(2) pneumoperitoneum was used to increase the intra-abdominal pressure to 30 mmHg (1 mmHg = 0.133 kPa) in 6 of 12 SAP animals thereafter and keep constant during the experiment. The investigation period was 12 h. Heart rate, cardiac output (CO), central venous pressure (CVP), mean arterial pressure and pulmonary arterial wedge pressure (PAWP) were continuously measured with the aid of balloon tipped flow-directed catheter and electrocardiography monitor. Oxygen partial pressure of artery (PaO(2)), carbon dioxide partial pressure of artery (PaCO(2)), ScvO(2), base excess (BE), and blood lactic acid (LAC) were measured by acid-base analysis.

RESULTSIn the IAH group, CO decreased significantly at 12 h, CVP and PAWP increased significantly at 3 h, 6 h and 12 h compared with SAP group (all P < 0.05). Peak inspiration pressure increased immediately after pneumoperitoneum in the IAH group, to (50.2 ± 3.1) cmH(2)O (1 cmH(2)O = 0.098 kPa) and (49.8 ± 0.9) cmH(2)O at 6 h and 12 h respectively. The pH, PaO(2), ScvO(2) and BE showed a tendency to fall in the IAH group. PaCO(2) and LAC were increased significantly in the IAH group (all P < 0.05).

CONCLUSIONSThere were remarkable and relatively irreversible effects on global hemodynamics in response to sustained IAH of 12 h with the underlying condition of SAP. Abdominal decompression is beneficial for patients of SAP with IAH.

Animals ; Disease Models, Animal ; Female ; Hemodynamics ; physiology ; Intra-Abdominal Hypertension ; Male ; Pancreatitis, Acute Necrotizing ; physiopathology ; Swine

Objective To observe the clinical efficacy of modified Shehuang Ointment(mainly composed of Cnidii Fructus,Phellodendri Chinensis Cortex,and Zanthoxyli Pericarpium)for the treatment of facial seborrheic dermatitis(SD).Methods Seventy-two patients with facial SD were randomly divided into observation group and control group,with 36 patients in each group.Both groups of patients were given oral use of Acrivastine Capsules and Vitamin B6 Tablets,and additionally,the observation group was given topical application of modified Shehuang Ointment and the control group was given topical application of 2%Ketoconazole cream.The course of treatment covered 4 weeks.The changes of clinical symptom scores and dermatology life quality index(DLQI)scores in the two groups were observed before and after treatment,and the clinical efficacy and safety of the two groups were also evaluated.Results(1)After 4 weeks of treatment,the total effective rate of the observation group was 88.89%(32/36),and that of the control group was 72.22%(26/36).The intergroup comparison showed that the efficacy of the observation group was significantly superior to that of the control group,and the difference was statistically significant(P＜0.05).(2)After treatment,the clinical symptom scores of erythema,scales,grease,rash area,itchiness and other clinical symptoms of the patients in the two groups were significantly decreased compared with those before treatment(P＜0.05),and the clinical symptom scores of the observation group were significantly lower than those of the control group,the differences being statistically significant(P＜0.05).(3)After treatment,the DLQI scores of patients in the two groups were significantly lower than those before treatment(P＜0.05),and the DLQI scores in the observation group were significantly lower than those in the control group after treatment,the difference being statistically significant(P＜0.05).(4)During the treatment period,no significant adverse reactions occurred in the two groups of patients,with high safety.Conclusion The conventional western medicine treatment combined with topical application of modified Shehuang Ointment exerts certain effect in the treatment of facial SD,which can effectively relieve the clinical symptoms and improve the quality of life of patients.

OBJECTIVETo compare the clinical effectiveness of percutaneous US- or CT-guided drainage and laparotomy for patients with infective pancreatic necrosis.

METHODSData of 90 infective pancreatic necrosis patients admitted from January 2008 to December 2009 were included. They were divided into two groups by the different treatment choices. Twenty-seven patients in the percutaneous group received percutaneous US- or CT-guided drainage as first choice. After that a sump suction apparatus was applied for controlled drainage. If no improvement was achieved after 3 days, they would received operation soon. While patients in the laparotomy group received surgical drainage at the time when the diagnosis of infected pancreatic necrosis was confirmed. Continuous drainage was also applied for these ones.

RESULTSThe percutaneous group had a significant low rate of reoperation (7.1% vs. 14.3%, P < 0.05) and postoperative residual abscesses (7.1% vs. 28.6%, P < 0.05). Furthermore, 48.1% of patients in percutaneous group successfully avoid laparotomy. In the regard of complications, the percutaneous group presented lower incidence of both single organ dysfunction (7.4% vs. 28.6%, P < 0.05), intestinal fistula (7.4% vs. 27.0%, P < 0.05) and long-term complications (3.7% vs. 22.2%, P < 0.05). In addition, the percutaneous group costed less medical resources as evidenced by shorter ICU duration (21.2 ± 9.7 vs. 28.7 ± 12.1, P < 0.01), shorter hospital duration (48.2 ± 12.5 vs. 59.6 ± 17.5, P < 0.05) and less expenditure (191 762 ± 5892 vs. 341 689 ± 10 854, P < 0.05).

CONCLUSIONSPercutaneous drainage can effectively lower the surgical rates and the rates of complications and reoperations in patients with infective pancreatic necrosis. Besides that, it could also reduce the cost of medical resources.

Adult ; Drainage ; methods ; Female ; Humans ; Laparotomy ; Male ; Middle Aged ; Pancreatitis, Acute Necrotizing ; surgery ; Retrospective Studies ; Treatment Outcome

Objective: To explore the clinical features, the serotype distribution and drug resistance of the isolates in patient with invasive pneumococcal disease (IPD). Methods: By retrieving the laboratory information system in 18 children′s hospitals from 2012 to 2017, the children with IPD were enrolled. Streptococcus pneumoniae (Spn) must be isolated from the sterile sites (blood, cerebrospinal fluid, hydrothorax and joint effusion etc.). The clinical characteristics, serotype, drug resistance, treatment and prognosis were reviewed and analyzed. According to the telephone follow up results, the patients were divided into death group and recovered group. The index as an independent risk factor of mortality was demonstrated by multivariate logistic regression analysis. Results: There were 1 138 children with IPD, including 684 male and 454 female. The proportion of male to female was 1.5∶1. The age ranged from one day to 16 years. The median age was 1 year 3 month. The majority was under 5 years of age (89.3%, n= 1 016), especially under 2 years of age (61.9%, n=704). In all cases, 88.2% (n=1 004) were community acquired infection. The infections included meningitis (n=446, 39.2%), pneumonia with bacteremia (n=339, 29.8%), and bacteremia without focus (n=232, 20.4%). Underlying diseases were found in 242 cases (21.3%). Co-infections were determined in 62 cases (5.4%) with mycoplasma, 27 cases (2.4%) with adenovirus and 34 cases with influenza virus (3.0%). The penicillin insensitivity (PNSP) rates in meningitis and non-meningitis isolates were 69.5% (276/397) and 35.9% (221/615), respectively. There were 81 strains serotyped, in which 93.8% (76/81) were covered by 13-valent protein-polysaccharide conjugate vaccine (PCV13). In the 965 patients who were followed up by phone call, 156 cases (16.2%) were confirmed dead. The independent risk factors for the death were under 2 years of age (OR=2.143, 95%CI 1.284-3.577, P=0.004), meningitis (OR=3.066, 95%CI 1.852-5.074, P<0.01), underlying disease (OR=4.801, 95%CI 2.953-7.804, P<0.01), septic shock(OR=3.542, 95%CI 1.829-6.859, P<0.01), disseminated intravascular coagulation (DIC) (OR=4.150, 95%CI 1.468-11.733, P=0.007), multiple organ failure (OR=12.693, 95%CI 6.623-24.325, P<0.01) and complications of central nervous system (OR=1.975, 95%CI 1.144-3.410, P=0.015). Conclusions Most children with IPD were under 5 years of age, having underlying diseases and acquired the infection in community. The independent risk factors for death were under two years old, meningitis, underlying diseases and multiple organ failure. The problem of drug resistance was severe. The universal immunization of PCV13 would be effective to prevent IPD in Chinese children.

OBJECTIVES: To explore the effect of acupuncture and moxibustion on intestinal flora in the rats with diarrhea-predominant irritable bowel syndrome (IBS-D) based on 16S rDNA technique. METHODS: Ten rats were randomized from 58 SPF-grade male SD rats to be the blank group. The remained 48 rats were prepared to be IBS-D models by the modified method of acetic acid enema combined with binding tail-clip stress. Forty successfully-modeled rats were randomly divided into a model group, an acupuncture group, a moxibustion group and a western medication group, with 10 rats in each one. In the acupuncture group, the needle was inserted at bilateral "Zusanli" (ST 36) and remained for 15 min in each rat. In the moxibustion group, the suspending moxibustion was delivered at bilateral "Zusanli" (ST 36) for 15 min. The rats in the western medication group were given pinaverium bromide suspension (10 mL/kg) by intragastric administration. The above interventions were performed once daily for consecutive 14 days. The body mass and the score of fecal trait were compared before and after modeling, as well as after intervention in each group. Fecal water content, diarrhea index and colon transit time (CTT) were measured after modeling and intervention in the rats of each group separately. After intervention, the colonic morphology of rats in each group was observed, and using 16S rDNA technique, the intestinal flora was detected. RESULTS: After modeling, compared with the blank group, the body mass and CTT were reduced (P<0.01); fecal trait scores, fecal water contents and diarrhea index increased (P<0.01) in the other 4 groups. After intervention, the body mass and CTT of the rats decreased (P<0.01), and fecal trait score, fecal water content and diarrhea index increased (P<0.01) in the model group compared with those in the blank group. In the acupuncture group, the moxibustion group and the western medication group, when compared with the model group, the body mass and CTT were elevated (P<0.01), while fecal trait scores, fecal water contents and diarrhea index declined (P<0.01). Compared with the western medication group, fecal water content decreased in the acupuncture group and the moxibustion group (P<0.05), while CTT increased in the acupuncture group (P<0.01), the body mass increased and fecal trait score was dropped in the moxibustion group (P<0.05). The colonic mucosa structure was clear and complete, and there was no obvious inflammatory cell infiltration in the blank group. The mild interstitial edema of intestinal mucosa was presented with the infiltration of few inflammatory cells in the model group. There was the infiltration of few inflammatory cells in the mucosa of the acupuncture group, the moxibustion group and the western medication group. Compared with the blank group, the indexes of Richness, Chao1, ACE and Shannon decreased in the model group (P<0.05). Indexes of Richness, Chao1 and ACE increased in the acupuncture group and the moxibustion group (P<0.05), and the Richness index in the western medication group increased (P<0.05) when compared with those in the model group. The relative abundance of Bacteroidetes, Proteobacteria and Prevotella increased (P<0.05), and that of Firmicutes and Muribaculaceae decreased (P<0.05) in the model group compared with those in the blank group. When compared with the model group, the relative abundance of Bacteroidetes, Proteobacteria and Prevotella was reduced (P<0.05), while that of Firmicutes and Muribaculaceae increased (P<0.05) in the acupuncture group, the moxibustion group and the western medication group; and that of Actinobacteria and Bifidobacterium increased in the acupuncture group and the moxibustion group (P<0.05). Compared with the blank group, the relative abundance of lipopolysaccharide (LPS) biosynthesis was elevated (P<0.05), and that of folate biosynthesis, lipoic acid metabolism, zeatin biosynthesis, ubiquinone and other terpenoid quinone biosynthesis decreased (P<0.05) in the model group. The relative abundance of LPS biosynthesis was dropped (P<0.05), and that of folate biosynthesis, lipoic acid metabolism, zeatin biosynthesis, ubiquinone and other terpenoid quinone biosynthesis increased (P<0.05) in the acupuncture group, the moxibustion group and the western medication group compared with those of the model group. CONCLUSIONS Either acupuncture or moxibustion can relieve the symptoms of IBS-D and protect intestinal mucosa, which may be associated with regulating the structure of intestinal flora and promoting nutrient metabolism and biosynthesis.
Rats ; Male ; Animals ; Irritable Bowel Syndrome/therapy* ; Moxibustion/methods* ; Rats, Sprague-Dawley ; Gastrointestinal Microbiome ; Lipopolysaccharides ; Thioctic Acid ; Ubiquinone ; Zeatin ; Acupuncture Therapy ; Diarrhea/therapy* ; Terpenes ; Water ; Folic Acid ; Acupuncture Points

This study was purposed to comparatively analyze the early T-lymphocyte subsets and T-cell receptor excision cycles (TREC) reconstruction in recipients with hematologic malignancies after myeloablative unrelated cord blood transplantation (UCBT) and sibling donor bone marrow and/or peripheral blood stem cell transplantation (BMT/PBSCT). The peripheral blood T lymphocyte subsets were detected using flow cytometry and TREC were detected using real-time quantitative PCR for 40 patients with hematologic malignancies in the first six months after myeloablative allogenic hematopoietic stem cell transplantation. The results showed that in the first month after transplantation, the absolute counts of CD3(+), CD3(+) CD4(+), CD3(+) CD8(+) cells were lower significantly in the UCBT group than those in the BMT/PBSCT group. And later the absolute counts of CD3(+), CD3(+)CD4(+), CD3(+)CD8(+) cells were not different between two groups. The ratio of CD3(+)T subset in the peripheral blood lymphocytes of the UCBT recipients was lower, but the difference was not statistically significant within 2 months after transplantation. The ratio of CD3(+)CD4(+) cells in the patients received the UCBT and BMT/PBSCT decreased obviously since engraftment happened. The CD3(+)CD4(+) cells on the 2 months after transplantation fell to the lowest level, then gradually increased, but did not reach to the normal level until 6 months after transplantation. CD3(+)CD8(+)cells were well reconstituted, rising to normal at the engraftment after transplantation, with a low CD4(+): [KG-*2] CD8(+) ratio over the first 6 months after transplantation. Compared with the BMT/ PBSCT group, the naive T cells (CD3(+)CD4(+)CD45RA(+)CD62L(+)) were more in the first month after transplantation and the terminally differentiated effector memory T cells (CD3(+)CD4(+)CD45RA(+)CD62L(-)) were more at the 3 month after transplantation in the UCBT group, and those were significantly more than the normal control group. TREC were lower and did not recovered until 6 months after transplantation in the recipients of the two groups. It is concluded that compared with sibling donor's BMT/PBSCT, early T cell reconstitution significantly delayed after UCBT, but the terminally differentiated effector memory T cells are higher after transplantation, and thus play a anti-infective or anti-leukemia role.
Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Fetal Blood ; transplantation ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Receptors, Antigen, T-Cell ; immunology ; T-Lymphocyte Subsets ; immunology ; Young Adult

Animals ; Biological Assay ; Nucleic Acid Amplification Techniques ; Phlebovirus/isolation & purification* ; Psychodidae/virology* ; Recombinases/metabolism* ; Reverse Transcription