OBJECTIVE To evaluate the in vitro inhibitory effects of antibacterials in combination carbapenem against multiresistant Pseudomonas aeruginosa.METHODS The protocol was designed by checkboard method and the MIC of the two antibiotics in combination was determined by broth dilution method.FIC was calculated according to MIC results.RESULTS The MIC50 and MIC90 of eight kinds of antibiotics were lower to 1-16 times and 1-64 times,respectively.The synergistic and additive antibacterial activity was seen in ceftazidime with amikacin,piperacillin with tobramycin and azlocillin with sulbactam.The lomefloxacine with ampicillin and lomefloxacine with ceftriaxone were not correlated.CONCLUSIONS Antibacterial activity of ceftazidime combined with amikacin against carbapenem-multiresistant P.aeruginosa is the best.

Objective To investigate the effect of small interfering RNA (siRNA) targeting cytoplasmic domain of tissue factor on apoptosis of vascular endothelial cells. Methods Specific siRNA targeting cytoplasmic domain of tissue factor were designed, and synthetic oligos were inserted into plasmid DNA. The siRNA constructs were transfected into human umbilical vascular endothelial cells (HUVEC) with liposome. The HUVEC were transfected with the constructs encoding siRNA Ⅰ, siRNA Ⅱ and pcDNA~(TM)6.2 GW/-miR plasmid separately. The transfected HUVEC were mixed with CD8~+ T lymphocytes. The apoptotic rate of tranfected HUVEC mixed with lymphocytes was analyzed by flow cytometry. Magnetic beads were used to measure PT of the supematant in the mixed lymphocytes culture. Results The siRNA constructs were confirmed by DNA sequence analysis. The apoptotic rate of HUVEC transfected with siRNA Ⅰ and Ⅱ plasmids was decreased significantly as compared with the empty control group (P<0.01). The apoptosis rate of HUVEC transfected with siRNA Ⅰ plasmid was lower than that of HUVEC transfected with siRNA Ⅱ plasmid (P<0.05). APTT of the culture supernatants in the three transfection groups was lower in the control groups (P <0.05), but there was significant difference among the three transfection groups. Conclusion The siRNA targeting cytoplasmic domain of tissue factor were successfully constructed, siRNA can protect HUVEC, and reduce the apoptotic rate of endothelial cells in mixed lymphocyte reaction without influencing the coagulation function.

The expression of paired immunoglobin-like receptors A (PIR-A) and B (PIR-B) and their relationship with tolerogenic dendritic cells (T-DC) in mice were investigated. The mouse DCs line, DC2.4 cells were cultured with the recombinant murine interleukin-10 (IL-10) and recombinant human transforming growth factor beta1 (TGF-beta1) respectively to develop the T-DC and stimulated with lipopolysaccharide (LPS) for 48 h to induce the mature dendritic cells (LPS-DC). Special small interfering RNAs (siRNA) molecule for PIR-B was chemically synthesized and transfected into DC2.4 cells (Si-DC) by lip2000. The expression of PIRs on DC2.4 cells were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), flow cytometry (FCM) and Western blot. Realtime reverse transcriptase-polymerase chain reaction (Realtime-PCR) was applied for measurement of PIR-A and CD80, CD86, MHC-II mRNA expression. The allogeneic stimulating capacity of DCs was measured by mixed lymphocyte reaction (MLR) using (3)H-thymidine incorporation test. The concentration of IFN-gamma in supernatants of MLR from distinct groups was analyzed by ELISA. The results showed that PIR positive rate was (28.65+/-8.12)% examined by FCM on DC2.4 cells. PIR positive rate was increased dramatically to (54.21+/-6.34)%, (58.78+/-4.70)%, (48.24+/-6.75)% respectively for IL-10, TGF-beta1 and LPS induction (P<0.01), but there was no significantly different among the three groups (P>0.01). The semi-quantitative RT-PCR and Western blot revealed that IL-10 and TGF-beta1 induced the higher PIR-B level and lower PIR-A level. On the contrary, the LPS down-regulated the PIR-B expression and up-regulated the PIR-A expression. Realtime PCR examination demonstrated that PIR-A and co-stimulating molecules such as CD80, CD86 and MHC-II were increased significantly after stimulation with LPS. Compared with the DC2.4 cells and the LPS-DC, the T-DCs inhibited alloactivated T cell proliferation and down-regulated the IFN-gamma secretion in MLR supernatant. Si-DC promoted the T cell proliferation (P<0.01) and enhanced the IFN-gamma secretion (P<0.01). It was concluded that up-regulating the PIR-B and down-regulating the PIR-A expression were the general feature of phenotype and constructed the new targets for dendritic cells to acquire immune tolerance in mice. Overexpression of PIR-B can inhibit the up-regulation of the PIR-A, CD80, CD86 and MHC-II expression, which might be the molecular mechanism for the T-DC.

Objective To investigate the clinical value of percutaneous gallbladder drainage in the treatment of severe acute pancreatitis(SAP). Methods A total of 65 patients treated for SAP in our hospital between January 2014 and April 2017 were analyzed retrospectively. The patients were divided into a gallbladder puncture group and a control group. Follow-up was performed for at least 6 months to monitor mortality and the incidence of complications, including pancreatic abscess, pseudocyst, renal failure, respiratory failure, heart failure, gastrointestinal bleeding, sepsis, and disseminated intravascular coagulation (DIC), The differences in mortality and complication rates between the two groups were statistically analyzed. Results Mortality in the gallbladder puncture group was significantly lower than in the control group (P ＜ 0. 05); the incidence of renal failure, respiratory failure, heart failure, gastrointestinal bleeding, and sepsis in the gallbladder puncture group was lower than in the control group (P ＜ 0. 05); the incidence of pancreatic abscess and pseudocyst in the gallbladder puncture group was similar to that in the control group, showing no significant difference (P ＞ 0. 05); the incidence of DIC in the gallbladder puncture group was lower than in the control group, but the difference was not statistically significant (P ＞ 0. 05). Conclusion Percutaneous gallbladder drainage can effectively reduce the incidence of renal failure, respiratory failure, heart failure, gastrointestinal bleeding, and sepsis in SAP, thereby reducing mortality. However, the incidence of DIC, pancreatic abscess, and pseudocyst is not reduced.

The expression of paired immunoglobin-like receptors A (PIR-A) and B (PIR-B) and their relationship with tolerogenic dendritic cells (T-DC) in mice were investigated. The mouse DCs line, DC2.4 cells were cultured with the recombinant murine interleukin-10 (IL-10) and recombinant human transforming growth factor β1 (TGF-β1) respectively to develop the T-DC and stimulated with lipopolysaccharide (LPS) for 48 h to induce the mature dendritic cells (LPS-DC). Special small interfering RNAs (siRNA) molecule for PIR-B was chemically synthesized and transfected into DC2.4 cells (Si-DC) by lip2000. The expression of PIRs on DC2.4 cells were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), flow cytometry (FCM) and Western blot. Realtime reverse transcriptase-polymerase chain reaction (Realtime-PCR) was applied for measurement of PIR-A and CD80, CD86, MHC-Ⅱ mRNA expression. The allogeneic stimulating capacity of DCs was measured by mixed lymphocyte reaction (MLR) using 3H-thymidine incorporation test. The concentration of IFN-γ in supernatants of MLR from distinct groups was analyzed by ELISA. The results showed that PIR positive rate was (28.65±8.12)% examined by FCM on DC2.4 cells. PIR positive rate was increased dramatically to (54.21±6.34)%, (58.78±4.70)%,(48.24±6.75)% respectively for IL-10, TGF-β1 and LPS induction (P＜0.01), but there was no significantly different among the three groups (P＞0.01). The semi-quantitative RT-PCR and Western blot revealed that IL-10 and TGF-β1 induced the higher PIR-B level and lower PIR-A level. On the contrary, the LPS down-regulated the PIR-B expression and up-regulated the PIR-A expression.Realtime PCR examination demonstrated that PIR-A and co-stimulating molecules such as CD80,CD86 and MHC-Ⅱ were increased significantly after stimulation with LPS. Compared with the DC2.4 cells and the LPS-DC, the T-DCs inhibited alloactived T cell proliferation and down-regulated the IFN-γ secretion in MLR supernatant. Si-DC promoted the T cell proliferation (P＜0.01) and enhanced the IFN-γ secretion (P＜0.01). It was concluded that up-regulating the PIR-B and down-regulating the PIR-A expression were the general feature of phenotype and constructed the new targets for dendritic cells to acquire immune tolerance in mice. Overexpression of PIR-B can inhibit the up-regulation of the PIR-A, CD80, CD86 and MHC- Ⅱ expression, which might be the molecular mechanism for the T-DC.

Animals ; Brain ; physiology ; Consciousness ; physiology ; Humans ; Mice ; Neurotransmitter Agents ; physiology ; Optics and Photonics ; Synaptic Transmission ; physiology

Periarticular fracture of the shoulder is a common type of fractures in the elderly. Postoperative adverse events such as internal fixation failure, humeral head ischemic necrosis and upper limb dysfunction occur frequently, which seriously endangers the exercise and health of the elderly. Compared with the fracture with normal bone mass, the osteoporotic periarticular fracture of the shoulder is complicated with slow healing and poor rehabilitation, so the clinical management becomes more difficult. At present, there is no targeted guideline or consensus for this type of fracture in China. In such context, experts from Youth Osteoporosis Group of Chinese Orthopedic Association, Orthopedic Expert Committee of Geriatrics Branch of Chinese Association of Gerontology and Geriatrics, Osteoporosis Group of Youth Committee of Chinese Association of Orthopedic Surgeons and Osteoporosis Committee of Shanghai Association of Chinese Integrative Medicine developed the Chinese expert consensus on the diagnosis and treatment of osteoporotic periarticular fracture of the shoulder in the elderly ( version 2023). Nine recommendations were put forward from the aspects of diagnosis, treatment strategies and rehabilitation of osteoporotic periarticular fracture of the shoulder, hoping to promote the standardized, systematic and personalized diagnosis and treatment concept and improve functional outcomes and quality of life in elderly patients with osteoporotic periarticular fracture of the shoulder.