OBJECTIVETo study the effects of sacral nerve root electrostimulation (SNS) on the colon function and its mechanisms in rats with spinal cord injury (SCI).

METHODSOne hundred and four Wistar rats were divided into three groups: A, B and C. A group ( n = 24) was divided into three subgroups (n = 8) for studying the bioelectricity: Normal group (NG), SCI group (SCI) and SCI group with SNS(SNS); B group( n = 24) was divided into three subgroups( n = 8) for studying the colon motility: NG, SCI and SNS. C group( n = 56) were divided into three groups for studying the change of morphology and neurotransmitters(SP and VIP): NG (n = 8), SCI (n = 24), and SNS (n = 24) . In SCI and SNS, included of three subgroups: 24, 48, 72 h after spinal cord injury (n = 8).

RESULTSIn SCI group, the activity of bioelectricity in proximal and distal colon was reduced; the colon motility was lessened, and colon mucosa appeared different degree of damage; cell-cell connections between intestinal epithelial cells were destroyed. The expressions of substance P(SP) and vasoactive intestinal peptide (VIP) in colon were decreased obviously. SNS was found to activate the bioelectricity, promote the colon motility, improve the intestinal mucosal, and increase the expressions of SP and VIP. Conclusion: SNS can activate the peristalsis, rehabilitate the motility of denervated colon, protection of the intestinal mechanical barrier between intestinal epithelial cells and tight junction, rebuild the colon function through activating the bioelectricity and increase the expressions of SP and VIP.

Animals ; Colon ; physiopathology ; Electric Stimulation Therapy ; Epithelial Cells ; drug effects ; Intestinal Mucosa ; drug effects ; Lumbosacral Region ; innervation ; Neurotransmitter Agents ; metabolism ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; therapy ; Substance P ; metabolism ; Vasoactive Intestinal Peptide ; metabolism

OBJECTIVETo observe the ultrastructural change of the route of gut bacterial translocation in a rat with spinal cord injury (SCI).

METHODSForty Wistar rats were divided into the following groups: control group and 3 SCI groups (10 in each group). The rats in the SCI groups were established SCI model at 24 h, 48 h, and 72 h after SCI. Small intestine mucous membrane tissue was identified and assayed by transmission electron microscope, scanning electron microscope and immunofluorescence microscopy.

RESULTSSmall intestine mucous membrane tissue in control group was not damaged significantly, but those in SCI groups were damaged significantly. Proliferation bacteria in gut lumen attached on microvilli. The extracellular bacteria torn the intestinal barrier and perforated into the small intestinal mucosal epithelial cell. The bacteria and a lot of particles of the seriously damaged region penetrated into the lymphatic system and the blood system directly. Some bacteria were internalized into the goblet cell through the apical granule. Some bacteria and particles perforated into the submucosa of the M cell running the long axis of M cells through the tight junctions. In the microcirculation of mucosa, the bacteria that had already broken through the microvilli into blood circulation swim accompanying with erythrocytes.

CONCLUSIONThe routes of bacterial translocation interact and format a vicious circle. At early step, the transcellular pathway of bacterial translocation is major. Following with the destroyed small intestine mucous, the routes of bacterial translocation through the lymphatic system and the blood system become direct pathways. The goblet cell-dendritic cell and M cell pathway also play an important role in the bacterial translocation.

Animals ; Bacteria ; Bacterial Translocation ; Epithelial Cells ; microbiology ; Goblet Cells ; microbiology ; Intestinal Mucosa ; microbiology ; pathology ; ultrastructure ; Intestine, Small ; microbiology ; pathology ; ultrastructure ; Microvilli ; microbiology ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; microbiology

Abstract: Objective To analyze a case of bloodstream infection caused by Ureaplasma urealyticum after abortion in Anxi County Hospital, so as to provide basis for the clinical diagnosis and treatment. Methods　The diagnosis of Ureaplasma urealyticum in this patient with bloodstream infection was retrospectively analyzed. The basic clinical data and laboratory diagnosis data were collected, including the characteristics of blood culture curve, Wright staining of culture medium, drug sensitivity of Mycoplasma liquid identification, colony characteristics of solid medium, and the conclusion of targeted DNA sequencing. Through the comprehensive analysis of the above data, the rapid diagnosis of this case can be realized by optimizing the detection and diagnosis process. Results　The clinical manifestations of this patient were fever of 38.5 ℃, CRP:14.85 mg/L, WBC:14.33×109/L, NET: 85.40%, PCT: 0.12 ng/mL, IL-6: 665.6 pg/mL, positive after 3 days of blood culture, no bacteria were found in Gram stain, and sand-like purple bacteria were observed after adding Wright's stain. After inoculation in blood agar, Mycoplasma solid and liquid medium, no colonies were grown in blood agar, after 48 h and 5 d. On Mycoplasma A7 agar, the edge of brown fried egg colony was striature, and it could be identified as Ureaplasma urealyticum with the Mycoplasma ID & AST panel, which was resistant to quinolones and spectinomycin, but sensitive to macrolides, tetracyclines and lincomycin. Subsequent targeted DNA sequencing results were also confirmed for Ureaplasma urealyticum. Before receiving the report, clinical experience treatment with ceftriaxone metronidazole was used to fight infection with negative bacilli and anaerobic bacteria. Mycoplasma was not treated with targeted treatment. After 3 days, the patient's body temperature returned to normal, inflammation index decreased, and the patient asked to be discharged. Conclusions　At present, there are few reports of bloodstream infection caused by Ureaplasma urealyticum, and the lack of clinical understanding can easily lead to misdiagnosis and missed diagnosis. In order to improve the detection rate of Mycoplasma in blood culture, it is necessary to optimize the detection procedure of blood culture and provide accurate diagnosis and treatment basis for clinical practice. However, it is clear from this case that Mycoplasma bloodstream infection cases are self-limited infection and can recover by themselves without targeted treatment in patients with normal immunity. Therefore, it is very important to protect the immunity of patients.

Objective To investigate the regulatory role of programmed death ligand-1 (PD-L1) on acute lung injury (ALI), and its molecular mechanism. Methods Twenty C57BL/6 male mice and 20 PD-L1 knock out male mice were collected, and they were divided into two groups by random number table, respectively: namely sham group and ALI group, 10 mice in each group. The model of ALI was reproduced by two-hit of hemorrhagic shock and sepsis, and the mice in sham group were only got bilateral femoral artery exposure and ligation without bleeding, cecal separation without ligation and perforation. The mice were sacrificed 24 hours after model reproduction, and the lung tissue and bronchoalveolar lavage fluid (BALF) were collected. The mRNA and protein expression levels of PD-L1 in the lungs were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and Western Blot. The pathological changes were observed with microscopy. The protein levels in BALF were determined. The granulocyte differentiation antigen 1 (Gr1) positive cells was determined by cytometry, and myeloperoxidase (MPO) activity in lung tissue was determined. The levels of proinflammatory factors interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and chemotatic factors keratinocyte chemoattractant (KC), macrophage inflammatory protein-2 (MIP-2) in lung homogenates and BALF were determined by enzyme-linked immunosorbent assay (ELISA). Results Compared with sham group, the mRNA and protein levels of PD-L1 in lung tissue of C57BL/6 mice in ALI group were significantly elevated [PD-L1 mRNA (2-ΔΔCt): 3.20±0.76 vs. 1.01±0.03, PD-L1 protein (A value): 0.98±0.16 vs. 0.15±0.04, both P < 0.05]. It was shown by light microscopy that the alveolar wall was thickened, congestive, edema and spot bleeding with a large number of inflammatory cell infiltration in the lung tissue of C57BL/6 mice in ALI group, and an obvious protein leakage was found in BALF (ng/L: 0.18±0.06 vs. 0.05±0.01, P < 0.05). The lung injury degree of PD-L1 knockout ALI mice was significantly less than that of C57BL/6 ALI mice, and the protein leakage was significantly reduced in BALF (ng/L: 0.11±0.02 vs. 0.18±0.06, P < 0.05). Compared with corresponding sham group, the number of Gr1 positive cells, MPO activity in lung tissue as well as the levels proinflammatory factors and chemotatic factors in lung tissue and BALF in ALI group were significantly increased. However, when compared with C57BL/6 ALI mice, above parameters in lung homogenates and BALF were significantly decreased in PD-L1 knockout ALI mice [number of Gr1 positive cells: (39.0±4.0)% vs. (45.0±8.0)%, MPO activity (U·μg-1·min-1): 2.85±0.62 vs. 4.52±1.16; lung IL-6 (ng/g): 461±111 vs. 728±28, TNF-α (ng/g): 1 123±175 vs. 1 500±327, KC (ng/g): 150±34 vs. 250±28, MIP-2 (ng/g): 1 263±468 vs. 1 763±323; BALF IL-6 (ng/L): 134±22 vs. 258±38, TNF-α (ng/L): 598±102 vs. 889±139, KC (ng/L): 934±286 vs. 1 258±336, MIP-2 (ng/L): 650±130 vs. 950±256; all P < 0.05]. Conclusion PD-L1 may play an important protective role in the immunological mechanism of ALI, which may be mediated by decreasing chemotactic factor KC and MIP-2 and mitigating neutrophil chemotaxis in lung tissue.

The medicinal plant resource reserve refers to the natural resources of medicinal plants in a certain time and a certain region within the scope of the volume. In recent years, with the demand of medicinal plant resources surging and the change of the environment and human intervention factors, the medicinal plant resources reserve had accelerated pace of change. It is the prerequisite and basis for the development and utilization of medical plants that how to quickly and accurately attain reserve of some medicinal plants resources, the selection of suitable and accurate estimating method is reliable basis and can guarantee medicinal plant reserve survey, and also is one of the key reserve investigation of success. This paper systematically summarized the estimation method of medicinal plants in recent 30 years, and discussed the basic principle, the estimation model of development and evolution, advantages and disadvantages and applicability, and it aimed to improve the accuracy about reserves survey of medicinal plant resources, and provide scientific and reliable support data to medicinal plants resources for sustainable development and utilization of resources.
China ; Conservation of Natural Resources ; Models, Statistical ; Plants, Medicinal ; growth & development

ObjectiveTo evaluate the functional changes of stunned myocardium before and after coronary artery bypass graft(CABG) treatment,and clear the meaning of revascularization which CABG has brought to patients with diffused vascular changes.MethodsA total of 36 patients with 99％ diffused coronary artery stenosis in left anterior descending branch underwent non-pump CABG treatment in the Department of Cardiothoracic Surgery the First affiliated hospital of Harbin Medical University.Real-time three-dimensional echocardiography (RT3DE) was repeatedly performed 1 week before operation and 10 days,1 month,and 3 months after CABG.Regional diastolic volumes,systolic volumes,ejection fractions,regional stroke volume to global diastolic volume and the values of abnormal segments before and after CABG were studied.ResultsOne week before operation and 10 days,1 month and 3 months after CABG,the differences of volumes between groups in the last phases of diastole and systolic were statistically significant in anterior wall basement segment,anterior septal basement segment,anterior wall intercalary segment,anterior septal intercalary segment,anterior wall of apex cordis and septation of apex cordis(F =3.51,3.55,4.08,4.05,2.98,3.01,all P ＜ 0.05; F =4.51,4.55,4.08,3.00,2.96,2.99,all P ＜ 0.05).The values of the six segments mentioned above,3 months after operation[(6.74 ± 1.23),(6.64 ± 1.21),(6.02 ± 1.10),(5.95 ± 1.09),(5.82 ± 1.06),(5.10 ± 0.93)ml; (2.74 ± 0.50),(2.69 ± 0.49),(2.51 ± 0.46),(2.32 ± 0.42),(2.36 ± 0.43),(2.03 ± 0.37)ml] were compared with those of 1 week before operation[(8.33 ± 1.52),(8.20 ± 1.50),(7.43 ± 1.36),(7.36 ± 1.34),(7.19 ± 1.31),(6.29 ± 1.15)ml; (4.94 ± 0.90),(4.85 ± 0.88),(4.53 ± 0.83),(4.18 ± 0.76),(4.25 ± 0.78 ),(3.65 ± 0.67)ml],the differences were statistically significant (all P ＜ 0.05); the differences between groups in regional ejection fractions,regional-global ejection fractions were statistically significant(F =4.56,4.88,4.28,3.15,2.93,2.88,P ＜ 0.01 or ＜ 0.05; F =5.56,5.28,4.98,5.15,3.03,2.78,P ＜ 0.01 or ＜ 0.05).Compared with 1 week before the operation, 1 month after the operation in regional ejection fractions,10 days,1 month in regionalglobal ejection fractions after the operation,4 segments of them were significantly improved(all P ＜ 0.05) and 3 months after operation,all the 6 segments had been improved significantly(all P ＜ 0.05).The maximum volume of the sum of group difference of the 6 segments and the 4 segments in the last phase of diastole was statistically significant(F =2.58,5.81,P ＜ 0.05 or ＜ 0.01 ),and the summation began to decrease 10 days after the operation.The values of 3 months after operation[ (36.27 ± 1.10),(25.35 ± 1.16)ml] were compared with that of 1 week before operation[ (44.80 ± 1.36),(31.32 ± 1.43)ml ] the difference was statistically significant (all P＜ 0.05).The maximum volume summafion comparisons of 6 segments and 4 segments in the last phase of systolic had statistical significance(F =5.77,5.57,all P ＜ 0.01 ),and 10 days after the operation,the summation began to decrease.The values of 1 month[(16.4 0 ± 0.48),(11.58 ±0.51 )ml],and 3 months after operation[ (14.65 ± 0.45),(10.26 ± 0.46)ml],were compared with those of 1 week before operation[ (26.40 ± 0.80),(18.50 ± 0.84)ml],the differences were statistically significant (all P ＜ 0.05).ConclusionsStunned myocardium can be improved through CABG in myocardium systolic,diastole function and ejection fractions of the relevant segments and all of this have proved that patients undergoing CABG revascularization can improve the heart function of the ischemic area.

Objective The immunotoxins generated by conjugating monoclonal antibody (mAb) and a certain toxin play an important and promising role in treating hematopoietic malignancies. However, most of the toxins used for the conjugation are toxic proteins, which are immunogenic in the patients. Norcantharidin (NCTD) is a small molecule toxin without immunogenicity, and thus has become a potential new drug for hematopoietic cancers. In this study, we prepared immunotoxin 2E8-NCTD by using the ZCH-4-2E8 cells produced in the laboratory of our hospital, and then detected its targeting effect against CD+19 lymphoid malignant Nalm-6 cells in vitro.Methods 2E8 mAb was obtained from mouse ascites and purified by gel chromatography. After its purity was checked by SDS-PAGE, immunotoxin 2E8-NCTD was generated by conjugating CD19 mAb with NCTD using activated ester method. The binding activity of the immunoconjugate to CD19 antigens on cell surface, and the expression levels of the CD19 antigens on Nalm-6 and K562 cells were determined by flow cytometry. The inhibitory effects of PBS, purified 2E8 mAb, NCTD, and immunotoxin 2E8-NCTD on the cell growth of either Nalm-6 or K562 cells were then compared.Results The purity of the 2E8 mAb was higher than 99% demonstrated by SDS-PAGE assay. 2E8 mAb was detected on the surface of 99.34% of the Nalm-6 cells, while on only 0.98% of the K562. The newly generated immunotoxin had a positive rate of 99.90% on the Nalm-6 with slightly reduced binding activity. Both 2E8-NCTD and NCTD significantly inhibited the growth of CD+19 Nalm-6 cells (P < 0. 001 ), while the purified 2E8 mAb did not show any significant influences on the growth of the same cell line ( P > 0.05 ). Meanwhile, no significant inhibitory effects on the CD-19 K562 cells were identified in the 2E8-NCTD, 2E8 mAb, or control groups, indicating a significant targeting effect of 2E8-NCTD against Nalm-6 cells.Conclusions The immunotoxin 2E8-NCTD can be synthesized by activated ester method. It has target killing effects on CD+19 Nalm-6 leukemia cells in vitro.

Objective To discuss the effect of psychological nursing on HIV/AIDS patients with anxiety and depression.Methods 40 HIV/AIDS patients with anxiety and depression were randomly divided into intervention group (20 cases) and control group (20 cases).Hamilton anxiety rating scale (HAMA) and Hamilton depression rating scale (HAMD) were used to evaluate the effect at 4 weeks and 3 months of the research.Results HAMA and HAMD scores of the intervention group at 4 weeks and 3 months were significantly lower than the control group [(15.30±4.12) vs (19.70±5.89),(13.00±2.51) vs (18.20±3.27),(5.70±2.57) vs (14.35 ±4.20),(7.50 ±3.19) vs (14.55 ±3.68),t =5.64,8.817,2.737,5.808;P ＜0.01].Conclusions Psychological nursing could greatly relieve the negative emotions of HIV/AIDS patients and promote treatment and nursing effect.

OBJECTIVETo investigate the role of Rictor/mTORC2 in the formation of blood testis barrier (BTB), testicular development, and spermatogenesis.

METHODSAmh Cre positive mice homozygous for rictor loxP with Sertoli cell specific deletion of rictor were obtained by cross breeding Amh Cre mice with rictor loxP mice. The histology of the reproductive organs, seminiferous tubules and epididymis of the transgenic mice was observed with HE staining. The cell subgroups of the germ cells in the seminiferous tubule were detected by flow cytometry with propidium iodide labeling. The expression levels of Ki 67 and separase were detected with immunofluorescence assay, and the expression levels of BTB associated proteins were detected with immunofluorescence and Western blotting.

RESULTSCompared with the control (Amh Cre, rictoror rictor) mice, the mice with Sertoli cell specific rictor deletion showed significantly decreased testicular weight and epididymis weight (P<0.05), significantly increased diploid cells (P<0.01), and decreased haploid cells (P<0.01) but comparable tetraploid cells and similar expression levels of Ki 67 and separase. The mice with rictor knockout also showed aberrant localization of BTB associated proteins, which were scattered over the whole seminiferous epithelium, but the expression levels of the protein remained stable.

CONCLUSIONRictor in testicular Sertoli cells is essential for maintaining BTB integrity and function and ensuring normal spermatogenesis in mice.

Two plasmid constructs, pcE2 and pcE3, containing 3' fragment of open reading frame 2 (ORF2,1163 bp) of hepatitis E virus (HEV) and full-length ORF3 (369 bp), were injected into bilateral tibialis of Swiss mice respectively，for three times (0, 2nd and 4th weeks) and observed the HEV IgG by ELISA. HEV IgG was induced after the injection of pcE2 or pcE3 or both, and the percentage of seraconversion was 100% after two weeks of the third injection. Compared with injection of either construct, the antibody titers were higher in the group with combined injection of two constructs.