Objective To investigate the clinical effect and immunologic function of Moxifloxacin combined with small dose of hormone in the treatment of Ventilator pneumonia in elder.Methods 64 cases of Ventilator pneumonia in our hospital were collected and randomly divided into experiment group and control group, 32 cases each.Two groups were given conventional treatment, the control group received Methylprednisolone Sodium Succinate 1 mg/kg qd, the experiment group was given Methylprednisolone Sodium Succinate 1 mg/kg qd, and Moxifloxacin Hydrochloride and Sodium Chloride Injection 400 mg qd.Two groups of patients were continuous treated for 10 days.After treatment,T lymphocyte subsets, NK cells, white blood cell count, C reactive protein, clinical symptoms disappeared time, mechanical ventilation time, ICU length of stay and mortality rate were compared. Results After treatment, the total effective rate in the experiment group 75% was higher than the control group 50%( P <0.05 ).The levels CD3 +, CD4 +, CD4 +/CD8 +and NK cell in two groups increased(P<0.05), levels of CD8 +decreased(P<0.05),levels of WBC, CPR and PCT decreased in the two groups(P<0.05), and compared with the control group, the levels CD3 +, CD4 +, CD4 +/CD8 +and NK cell in the experiment group were higher(P<0.05), levels of CD8 + were lower(P<0.05),levels of WBC,CPR and PCT were lower(P <0.05), the rales disappeared time, cough disappeared time, fever disappeared time were significantly shorter than the control group(P <0.05), the duration of mechanical ventilation and the length of hospital stay were significantly shorter(P<0.05).Conclusion Moxifloxacin combined with small dose of hormone in the treatment of Ventilator pneumonia in elder was significantly effective, and it can relieve inflammation, prevention of infection control, enhance immune function.

Objective To prepare the Env-pseudotyped subtype B HIV-1 with enhanced green fluorescent protein ( EG-FP) gene,explore HIV-1 infection mechanisms and develop feasible methods of identification .Methods The Env-pseudo-typed viruses were packaged in HEK293T cells by cotransfection, and the reporter gene and P24 protein were detected by PCR, Western blot and ELISA .Reporter gene amplification , viral titration assay and a single round of infection assay were performed after the env-pseudotyped viruses infected HIV-1 permissive cell .Results and Conclusion A generation and identification method of the pseudotyped HIV-1 was established . The Env-pseudotyped subtype B HIV-1 has been prepared, which is able to infect SupT1 and TZM-bl cells through infection assay .

Objective To investigate the expression and significance of transforming growth factor-β1 (TGF-β1) gene in bone marrow mesenchymal stem cell (BMMSC) derived from patients with multiple myeloma (MM).Methods BMMSC of 7 MM patients and 10 patients with iron deficiency anemia were cultured in vitro.The morphology of BMMSC was observed and the growth curve was portrayed according to the daily results of BMMSC proliferation.Total RNA was extracted from BMMSCs and transcription of TGF-β1 gene in BMMSC was measured by reverse transcription-PCR.Results The proliferative activity of BMMSC was not significantly different between the two groups,but expression of TGF-β1 gene of BMMSC was higher in MM patients (0.01241±0.00419) than the control group (0.00122±0.00030) (t =3.218,P ＜ 0.05).Conclusion The abnormally high expression of TGF-β31 gene in BMMSCs could contribute to the pathogenesis of MM.

Objective:To explore the molecular mechanism for bone mass loss caused by staphylococcus aureus infection.Methods:Thirty 8-week-old male C57BL/6 mice were randomly divided into 3 groups ( n=10): control, infection and infection+JAK inhibitor (JAKi) ones. The mice were killed 2 weeks later for sampling from the femur and tibia. Micro-CT reconstruction was performed for analyses of BV/TV, Tb.N, Tb.Th and Tb.Sp to detect changes in bone mass; OCN immunohistochemistry and Goldner's trichrome staining were used to quantify osteoblasts; TRAP staining was used to quantify osteoclasts; the GSE166522 data set was downloaded and analyzed to explore the relationships between staphylococcus aureus infection and bone cell senescence and JAK/STAT pathway. Senescence β-Galactosidase staining, Osterix and P16 immunofluorescence colocalization were used to observe the changes in number of senescent cells. Results:MicroCT results showed a statistically significant difference in the loss of cancellous bone in the target area in the infection group compared with the control group ( P<0.05). The results of osteocalcin immunohistochemistry and Goldner's trichrome staining indicated that the number of osteoblasts in the infection group was significantly reduced ( P<0.05). TRAP staining indicated no significant difference in the number of osteoclasts between the infection and control groups ( P>0.05). Bioinformatics analysis found that staphylococcus aureus infection caused bone cell senescence and the JAK/STAT pathway was activated after the infection. Senescence β-Galactosidase staining suggested that senescent cells increased in the infection group compared with the control group. The number of Osterix and P16 positive senescent osteoprogenitor cells in the infection group was increased significantly compared with the control group. The number of senescent osteoprogenitor cells in the infection+JAKi group was significantly reduced and the bone loss was partially reversed after treatment of JAK inhibitor, compared with the infection group. Conclusion:Staphylococcus aureus may induce osteoprogenitor cell senescence through the JAK/STAT pathway and eventually lead to bone mass loss.

Objective:To explore the effect of hydrogen sulfide (H 2S) on modulating the subunit Kir6.2 of adenosine triphosphate sensitive potassium channels via the cyclic guanosine monophosphate-dependent protein kinase (cGMP/PKG) signaling pathway in epileptic rat models. Methods:Sixty adult male SD rats were randomly divided into the following six groups (10 rats in each group) by random number table method: control, epileptic, H 2S donor, H 2S donor+epileptic, KT5823 (one inhibitor of the cyclic guanosine monophosphate-dependent protein kinase)+H 2S donor+epileptic, and glibenclamide (one inhibitor of the adenosine triphosphate sensitive potassium channels)+H 2S donor+epileptic groups. Except the control group, SD rats were intraperitoneally injected with plentylenetetrazole to make the kindling models and their behaviours were recorded including the latency period, the grade, and the duration of the first epileptic seizure according to the Racine′s standard. The waveforms of electroencephalogram (EEG) in hippocampus were also recorded during the seizure. The mRNA and protein levels of PKG and Kir6.2 in hippocampus were evaluated by Western blotting and quantitative real-time polymerase chain reaction, and the hippocampal concentrations of cGMP and phosphorylation of cyclic guanosine monophosphate-dependent protein kinase (p-PKG) were detected by enzyme linked immunosorbent assay. Results:Rats in the epileptic group showed Ⅳ-Ⅴ grade of epileptic seizure [4.500 (4.000, 4.875)], short latency period [(10.37±8.21) min] but long duration [(69.50±24.37) s] of seizure. Compared to the epileptic group, rats in the H 2S donor group showed Ⅱ-Ⅲ grade of epileptic seizure ( P=0.004), significantly longer latency period ( P<0.001), and shorter duration of seizure ( P<0.001). Compared to the H 2S donor+epileptic group, rats in the KT5823+H 2S donor+epileptic group showed Ⅲ-Ⅳ grade of epileptic seizures, significantly shorter latency period ( P<0.001), and longer duration of seizure ( P<0.001). The results of EEG showed that the wave patterns in the epileptic group were spike or sharp waves and the amplitudes were largest [(190.570±23.590) μV]. Compared with the epileptic group, amplitudes were reduced ( P<0.001) in the H 2S donor+epileptic group. PKG mRNA and PKG protein were expressed differently among all groups (PKG mRNA: n=5, H=26.714, P<0.001; PKG protein: n=5, F=30.597, P<0.001). Compared with the control group, the expression of both PKG mRNA and PKG protein was decreased (PKG mRNA: 1.000±0.001 vs 0.782±0.064, P=0.023; PKG protein: 0.550±0.037 vs 0.145±0.020, P=0.042) in the epileptic group. Besides, Kir6.2 mRNA and Kir6.2 protein were expressed differently among all groups (Kir6.2 mRNA: n=5, H=27.761, P<0.001; Kir6.2 protein: n=5, F=60.659, P<0.001). Compared with the control group, the expression of both Kir6.2 mRNA and Kir6.2 protein was decreased (Kir6.2 mRNA: 1.000±0.001 vs 0.897±0.033, P=0.004; Kir6.2 protein: 0.384±0.035 vs 0.215±0.016, P=0.024) in the epileptic group. And the concentrations of cGMP and p-PKG were decreased (cGMP: P<0.001; p-PKG: P<0.001) in the epileptic group. The results in the H 2S donor+epileptic group were up-regulated (PKG mRNA: P=0.047; PKG protein: P<0.001; Kir6.2 mRNA: P=0.011; Kir6.2 protein: P<0.001; cGMP: P<0.001; p-PKG: P<0.001) compared with the epileptic group. However, the results in the KT5823+H 2S donor+epileptic group were down-regulated (PKG mRNA: P=0.015; PKG protein: P=0.027; Kir6.2 mRNA: P=0.013; Kir6.2 protein: P=0.017; cGMP: P=0.005; p-PKG: P<0.001) compared with the H 2S donor+epileptic group. Conclusion:A possible mechanism is that H 2S prevents the epileptic seizure from modulating the subunit Kir6.2 of ATP sensitive potassium channels via the cGMP/PKG signaling pathway.

Objective To observe and analyze the clinical treatment effect of chronic heart failure patients with renal insufficiency:Methods Retrospective analysis our hospital inter-treated 80 patients with chronic heart failure patients complicated renal insufficiency in January 2004 ～ June 2008 cardiac function of patients with grade,the use of inotropie agent drugs,angiotensin converting enzyme inhibitors and adrenergic receptor binder chugs, 15-blocker treatment and other methods. Results Cardiac function grade Ⅳ renal dysfunction in the patient's prognosis than obvious cardiac function grade Ⅲ renal dysfunction patients worse,it is significant difference (P< 0.05). Conclusion Renal insufficiency and prognosis of heart failure is closely related to renal dysfunction in heart failure patients to a reasonable selection of appropriate drugs,the drug application process can foresee possible adverse reactious,to closely monitor patient and make efforts to preserve renal function.

Objective To investigate the protective effect of high-pressure carbon monoxide for preservation of ex vivo rabbit heart graft in comparison with the conventional HTK cardioplegic solution preservation. Methods Heart grafts isolated from 85 New Zealand rabbits were randomly divided into Naive group (n=5), HTK group (n=40) and CO group (n=40). The grafts underwent no preservation procedures in Naive group, preserved at 4 ℃ in HTK cardioplegic solution in HTK group, and preserved at 4℃in a high-pressure tank (PO2:PCO=3200 hPa:800 hPa) in CO group with Krebs-Henseleit solution perfusion but without cardioplegic solution. After preservation for 2, 4, 6, 8, 10, 14, 18, and 24 h, 5 grafts from the two preservation groups were perfused for 30 min with a modified Langendorff apparatus and examined for left ventricular systolic pressure (LVSP), left ventricular diastolic pressure (LVDP), arrhythmia score (AS), myocardial ultrestructure, and cardiac enzyme profiles. Results After preservation for 6 to 24 h, the cardiac enzyme profiles and systolic and diastolic functions were significantly better in CO group than in HTK group, but these differences were not obvious between the two groups after graft preservation for 2 to 4 h. Significant changes in the myocardial ultrastructures occurred in the isolated hearts after a 24-h preservation in both CO and HTK groups, but the myocardial damages were milder in CO group. Conclusion Preservation using high-pressure carbon monoxide can better protect isolated rabbit heart graft than the conventional HTK preservation approach especially for prolonged graft preservation.

The circadian clock is a generator of self-sustaining physiological and behavioral rhythms, which can be guided by external environmental factors, so as to synchronize biological behaviors with external environmental changes. The modern lifestyles make the human body incapable of synchronization to the external time with the circadian rhythm, and thus form a social jet lag. Non-alcoholic fatty liver disease (NAFLD) is a disorder closely related to metabolic abnormalities. The circadian clock is closely related to metabolic abnormalities and NAFLD and changes among them may be involved with feeding mode and ingredients, sleeping time, and intestinal flora. Molecules associated with the circadian clock are expected to become potential drugs for the treatment of NAFLD. This article mainly reviews the latest research progress of circadian clock and NAFLD.

Objective To investigate the protective effect of high-pressure carbon monoxide for preservation of ex vivo rabbit heart graft in comparison with the conventional HTK cardioplegic solution preservation. Methods Heart grafts isolated from 85 New Zealand rabbits were randomly divided into Naive group (n=5), HTK group (n=40) and CO group (n=40). The grafts underwent no preservation procedures in Naive group, preserved at 4 ℃ in HTK cardioplegic solution in HTK group, and preserved at 4℃in a high-pressure tank (PO2:PCO=3200 hPa:800 hPa) in CO group with Krebs-Henseleit solution perfusion but without cardioplegic solution. After preservation for 2, 4, 6, 8, 10, 14, 18, and 24 h, 5 grafts from the two preservation groups were perfused for 30 min with a modified Langendorff apparatus and examined for left ventricular systolic pressure (LVSP), left ventricular diastolic pressure (LVDP), arrhythmia score (AS), myocardial ultrestructure, and cardiac enzyme profiles. Results After preservation for 6 to 24 h, the cardiac enzyme profiles and systolic and diastolic functions were significantly better in CO group than in HTK group, but these differences were not obvious between the two groups after graft preservation for 2 to 4 h. Significant changes in the myocardial ultrastructures occurred in the isolated hearts after a 24-h preservation in both CO and HTK groups, but the myocardial damages were milder in CO group. Conclusion Preservation using high-pressure carbon monoxide can better protect isolated rabbit heart graft than the conventional HTK preservation approach especially for prolonged graft preservation.

Objective To investigate the effect of human bone marrow mesenchymal stem cells (BM-MSCs) stimulated by platelets in vitro on the metastasis of cancer cells.Methods The BM-MSCs were isolated and cultured in vitro and platelets from the peripheral blood of healthy persons were purified.The MSCs (control),platelets + MSCs,and platelets treated with culture media (CM) of SGC-7901 tumor cells + MSCs (T-platelets + MSCs) were cultured,respectively,and the MSCs and supernatants (MSCs-CM and SGC-7901-CM) were collected,respectively,after 24 hours.The expressions of markers of cancer-associated fibroblasts (CAF),such as α-SMA and Vimentin,were determined by Western-blotting.The immigration ability of BM-MSCs were analyzed by Transwell assay.The levels of P-selectin in platelets stimulated by MSCs-CM or SGC-7901-CM were detected with flow cytometry.The metastasis model of gastric cancer SGC-7901 cells was established in BALB/c nude mice by the injection of tail vein,and the tumor metastasis in vivo was observed.Results The expression levels of P-selectin in platelets stimulated by MSCs-CM ([21.37 ± 1.00] ％) or SGC-7901-CM ([31.4 ± 1.71] ％ were significantly higher than that in the control ([3.17 ± 0.40] ％,t =27.85 and 29.18,P ＜ 0.01).The expression levels of α-SMA and Vimentin in platelets + MSCs group (0.79 ± 0.08 and 0.88 ± 0.01) and T-platelets + MSCs group (0.90 ±0.06 and 0.96 ±0.04) were significantly higher than that in the control (0.64 ±0.02 and 0.75 ±0.05,t =2.96 and 6.45 forα-SMA,t =4.73 and 5.73 for Vimentin,P ＜0.01).The amounts of immigration cells in platelets + MSCs group (340.3 ±27.7) and T-platelets ± MSCs group (424.3 ± 17.6) were significantly higher than that in the control (220.7 ± 19.4,t =6.14 and 13.48,P ＜ 0.01).The in vivo experimental results showed that the metastatic foci in platelets ± MSCs group (4 ± 2) and T-platelets ± MSCs group (21 ± 4) were significantly higher than that in the control (0.33 ± 0.06,t =3.051 and 8.857,P ＜ 0.01).Conclusion Platelets promote the immigration and the enhanced tumor metastasis in vivo of BM-MSCs.