Objective:To investigate the mechanism of trigger point deactivation induced by pressing manipulation in a rat model and to explore its potential regulation of the inflammatory response through the extracellular signal-regulated kinase(ERK)/nuclear factor-κB(NF-κB)pathway. Methods:Fifty male Sprague-Dawley rats were randomly divided into a blank group,a model group,a pressing manipulation group,an ERK agonist group,and a pressing manipulation+ERK agonist group,with 10 rats in each group.Except for the blank group,rats in other groups were used to establish the trigger point rat model using the blunt blow combined with the eccentric exercise method.The pressing manipulation group underwent pressing manipulation intervention at the trigger points.The ERK agonist group received an injection of recombinant human epidermal growth factor via the tail vein.The pressing manipulation+ERK agonist group received interventions from both the pressing manipulation and ERK agonist groups.The pressure pain threshold(PPT)was measured by a mechanical pain threshold detector before and after the intervention.The histological changes were evaluated by hematoxylin-eosin staining after the intervention;the expression levels of ERK,phosphorylated ERK(p-ERK),NF-κB p65(p65),phosphorylated NF-κB p65(p-p65),and phosphorylated NF-κB inhibitor(p-IκB)were detected by Western blotting;the levels of interleukin(IL)-1β,IL-6,and tumor necrosis factor(TNF)-α were detected by enzyme-linked immunosorbent assay. Results:The PPT increased(P<0.05);the inflammatory cells disappeared;the ratios of p-ERK/ERK,p-p65/p65,and p-IκB/β-actin,also the levels of IL-1β,IL-6,and TNF-α all decreased in the pressing manipulation group after the intervention compared with the model group(P<0.05).The PPT decreased significantly(P<0.05),the inflammatory cell presence increased,and the ratios of p-ERK/ERK and p-p65/p65 were elevated(P<0.05);additionally,the levels of IL-6 and TNF-α were significantly higher in the pressing manipulation+ERK agonist group compared with the pressing manipulation group(P<0.05).The PPT was significantly lower(P<0.05),the inflammatory cell count was higher,the ratios of p-ERK/ERK and p-IκB/β-actin and the levels of IL-1β and TNF-α were significantly higher in the ERK agonist group compared with the pressing manipulation+ERK agonist group(P<0.05). Conclusion:Pressing manipulation can effectively alleviate inflammation and pain in trigger point model rats,potentially by inhibiting the ERK/NF-κB signaling pathway.

Objective:To evaluate the effect of perioperative transcutaneous electrical acupoint stimulation (TEAS) on postoperative cellular immune function in the patients undergoing posterior spinal internal fixation.Methods:Ninety patients, aged 40-70 yr, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, undergoing elective posterior spinal internal fixation in our hospital, were divided into 2 groups ( n=45 each) using a computer-generated table of random numbers: routine group and experiment group.Total intravenous anesthesia was used in routine group, while total intravenous anesthesia combined with TEAS was applied in experiment group.In experiment group, bilateral Zusanli and Sanyinjiao acupoints were stimulated with 2/15 Hz disperse-dense waves at the intensity that could be tolerated by patients at 30 min before induction of anesthesia, maintaining with 2/100 Hz disperse-dense waves from the end of induction until the end of operation at the same stimulation intensity before induction.Bilateral Neiguan and Taichong acupoints were stimulated for 30 min each time with 2/15Hz disperse-dense waves once a day at 1st-4th days after operation.In routine group, the electrodes were connected at the same time period, but no stimulation was given.Venous blood samples were collected before induction of anesthesia, at 1 h after surgery, and on 1st, 3rd, and 5th days after surgery, and the percentage of CD3 + , CD4 + , CD8 + T lymphocytes, CD4 + /CD8 + ratio, WBC count and percentage of neutrophils (NE%) were determined by flow cytometry, and the consumption of intraoperative anesthetics, use of postoperative analgesics, nausea and vomiting, dizziness, infection and length of hospital stay were recorded. Results:Compared with routine group, the total consumption and consumption index of remifentanil were significantly decreased, the percentage of CD3 + T lymphocytes was increased on 3rd and 5th postoperative days, the NE% was decreased on 1st postoperative day, and the incidence of dizziness was decreased ( P<0.05), and no significant change was found in the other indicators in experiment group ( P>0.05). Conclusions:Perioperative TEAS can improve postoperative cellular immune function and has a certain potential value in preventing postoperative infection in the patients undergoing posterior spinal internal fixation.

Objective:To summarize and analyze the clinical data of Chinese patients with colony-stimulating factor 1 receptor (CSF1R)-related leukoencephalopathy, and clarify the phenotypic and genetic characteristics of Chinese patients.Methods:Medical history of patients with CSF1R-related leukoencephalopathy diagnosed from April 1, 2018 to January 31, 2021 in the department of neurology of 22 hospitals in China was collected, and scores of Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment Scale (MoCA), magnetic resonance severity scale were evaluated. Group comparison was performed between male and female patients.Results:A total of 62 patients were included, and the male-female ratio was 1∶1.95. The age of onset was (40.35±8.42) years. Cognitive impairment (82.3%, 51/62) and motor symptoms (77.4%,48/62) were the most common symptoms. The MMSE and MoCA scores were 18.79±7.16 and 13.96±7.23, respectively, and the scores of two scales in male patients (22.06±5.31 and 18.08±5.60) were significantly higher than those in females (15.53±7.41 , t=2.954, P=0.006; 10.15±6.26, t=3.328 , P=0.003). The most common radiographic feature was bilateral asymmetric white matter changes (100.0%), and the magnetic resonance imaging severity scale score was 27.42±11.40, while the white matter lesion score of females (22.94±8.39) was significantly higher than that of males (17.62±8.74 , t=-2.221, P<0.05). A total of 36 CSF1R gene mutations were found in this study, among which c.2381T>C/p.I794T was the hotspot mutation that carried by 17.9% (10/56) of the probands. Conclusions:The core phenotypic characteristics of CSF1R-related leukoencephalopathy in China are progressive motor and cognitive impairment, with bilateral asymmetrical white matter changes. In addition, there exist gender differences clinically, with severer cognitive impairment and imaging changes in female patients. Thirty-six CSF1R gene mutations were found in this study, and c.2381T>C/p. I794T was the hotspot mutation.

Objective To evaluate the efficacy of haploidentical allogeneic hematopoietic stem cell transplantation (Haplo-HSCT) combined with third-party umbilical cord blood (UCB) infusion in treatment of high-risk lymphoblastic malignancies. Methods The clinical data of 20 patients with high-risk lymphoblastic malignancies who received Haplo-HSCT from April 2012 to April 2015 in Shanghai General Hospital were retrospectively analyzed, which were compared with the data from 15 patients who underwent matched unrelated donor HSCT (MUD-HSCT) or 14 matched sibling donor HSCT (MSD-HSCT) during the same period. The efficacy of Haplo-HSCT combined with UCB infusion in treatment of high-risk lymphoblastic malignancies was evaluated. The preparative regimen mainly consisted of teniposide, cyclophosphamide and total body irradiation (TBI). Graft versus host disease (GVHD) preparative regimen included cyclosporine and a short term of methotrexate. The patients who received Haplo-HSCT combined with UCB infusion and MUD-HSCT were treated with antithymocyte globulin (ATG). Results After the transplantation, one patient in MUD-HSCT group and one in MSD-HSCT group died within 21 days, and other patients were engrafted successfully. The median time of neutrophil engraftment was 13 days (10-18 d), 12 days (9-16 d) and 12 days (9-14 d) in Haplo-HSCT + UCB group, MUD-HSCT group and MSD-HSCT group, respectively; the median time of platelets engraftment was 11 days (9-18 d), 12 days (10-23 d) and 12 days (9-14 d), respectively. There were 10, 3, 3 cases of grade Ⅱ-Ⅳacute GVHD at day 100 in the three groups, respectively, and there were 6, 4, 3 cases of chronic GVHD in the three groups, respectively. The 2-year cumulative incidence of relapse was 40.6%, 66.2% and 26.7%, respectively. The predicted 2-year overall survival rate was 37.9%, 42.9% and 55.4%, respectively. All these data had no significant difference (all P＞ 0.05). Conclusion The efficacy of Haplo-HSCT combined with UCB infusion is similar to that of MUD-HSCT or MSD-HSCT in treatment of high-risk lymphoblastic malignancies, which should be recommended to the patients with high-risk lymphoblastic malignancies and without matched donors.

Objective: To evaluate the diagnostic value of bronchoalveolar lavage (BAL) for pulmonary complications in patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and its safety. Methods: Patients with pulmonary complications after allo-HSCT underwent BAL. Microbiological smears, culture, PCR of CMV-DNA, EBV-DNA and TB-DNA, macro genomes new generation sequencing (mNGS) techniques were performed to detect pathogens in BAL fluid (BALF) . Results: A total of 73 allo-HSCT patients with 86 times of pulmonary complications enrolled this prospective study. They underwent 132 times of BAL procedures. The clinical diagnoses of 88.4% cases were made based on BALF analysis. Of them, 67 cases (77.9%) had infectious pulmonary complications, including 29 cases (33.7%) of fungal infection, 18 cases (20.9%) of mixed infection, 11 cases (12.8%) of viral infection and 9 cases (10.5%) of bacterial infection. The other 9 cases (10.5%) of non-infectious pulmonary complications included 8 cases (9.3%) of idiopathic pneumonia syndrome (IPS) and 1 case (1.2%) of pulmonary infiltration of lymphoma. The diagnoses of the remaining 10 cases (11.6%) were not determined. The platelet counts of 33 patients were less than 50×10⁹/L before BAL. None of them developed severe bleeding complications during or after BAL. Transient fever occurred in 10 patients after BAL. Blood cultures showed staphylococcal bacteremia in them and anti-infection therapies were effective. No life-threatening complications occurred in all of the patients during or after BAL. Conclusion BALF analysis was informative for the diagnosis of pulmonary complication and safe for patients with pulmonary complications after allo-HSCT.

Objective To evaluate the efficacy of peripheral blood combined with cord blood model for haplo-hematopoietic stem cell transplantation and the occurrence,survival of complications in patients of different ages.Methods From January 2014 to December 31,2017,there were 50 patients undergoing haploid allogeneic hematopoietic stem cell transplantation in our department.There were 39 males and 11 females.The median age was 35 (9-67) years.The stratification was divided into 3 groups.In group A,17 patients were younger than 30 years old;in group B,19 patients were between 30 and 49 years old,and in group C,14 patients were not less than 50 years.No remission was assessed before transplantation in this group.On the morning of the reinfusion,the selection of a third-party umbilical cord blood for transfusion reduced the occurrence of GVHD.Peripheral blood was infusion in the afternoor.All patients were treated with ATG + CSA + shortterm MTX to prevent GVHD.Results Two patients died of infection prior to graft,4 (8.0％) patients were graft failure.The median time of ANC≥0.5 × 109/L (range) and platelet ≥20 × 109/L (range) in the other patients were 14d(10-22 d) and 20(11-186) d,individually.The median time of full donor chimerism(range)was 28d(14-42 d).Graft failure was occurred in one case (5.9％),two cases (10.5％) and one case (7.1％) in each group,with no statistically significant difference (x2 =0.282,P =0.868).With a median follow-up of 7.2 months (0.4-27.2 months),12 (24％) had aGVHD of Ⅱ-Ⅳ degrees,among them,6 cases (35.3％) in group A,5 cases (26.3％) in group B,1 case (7.1％) in group C had aGVHD of Ⅱ-Ⅳ degrees.There was no significant in the incidence of aGVHD in three groups (x2 =3.624,P =0.180).Twenty-nine (58％) patients had viral infections after transplantation.One patient in both group A and B relapsed,and there was no recurrence in group C.21 (42％) patients died and 29 (58％) patients survived.The predicted 2-year overall survival (OS) was 60.2％.In group C,the 2-year overall survival (OS) was 77.1％.Conclusion The haploidentical hematopoietic cell transplantation model of peripheral blood combined with third-party umbilical cord blood transfusion has a good outcome and prolonged survival time in high-risk elder patients.The use of suitable conditioning regimens did not increase the incidence of aGVHD and virus infection.

Objective: To evaluate the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for elderly patients with advanced myeloid neoplasm. Methods: From September 2014 to September 2017, 30 consecutive hospitalized 50-plus-year-old myeloid neoplasm patients were retrospectively analyzed. At the time of transplantation, 6 patients reached complete remission and the others remained no remission after treatment. The donors were identical sibling (12), matched unrelated (6) and haploidentical family member (12), respectively. 18 patients received RIC while 12 patients received MAC conditioning regiments consisted of Busulfan, cytarabine, fludarabine or clarithromycin±TBI, respectively. Results: Five patients died early in the conditioning stage, 24 patients successfully engrafted. The median time of neutrophil engraftment was 14(10-18) d, whereas platelet engraftment was 15(10-19) d. Six cases (25%) experienced aGVHD grades Ⅱ, 8 cases (32%) cGVHD, including moderate to severe cGVHD in 2 cases (8%). Seven, 7 and 5 cases developed CMV viremia, pneumonia and herpeszoster, respectively after transplantation, but no patients died of infections. The median follow-up time of the patients was 7(0.5-38) months. Twenty-one patients were still alive. The estimated 2 years OS and LFS were 62.5% (95% CI 39.2%-85.8%) and 59.2% (95% CI 26.9%-91.5%), respectively. Univariate analysis showed that HCT-CI was the only factor influencing OS. Conclusion Allogeneic hematopoietic stem cell transplantation could improve the survival of elderly patients with myeloid neoplasm.

Objective: To evaluate the efficacy of reduced-intensity conditioning allogeneic hematopoietic stem cell transplantation (RIC-allo-HSCT) for patients with myelofibrosis (MF). Methods: The clinical data of 10 patients with myelofibrosis (MF) who underwent RIC-allo-HSCT. Results: Of all 10 patients, 6 were male and 4 women, with a median age of 28.5 (22-54). Using fludarabine/busulfan plus total body irradiation (FB+TBI) pretreatment scheme based. Hematopoiesis reconstitution was achieved in 9 patients (90%). The median time of neutrophil and platelet engraftment was 13.5 (10-22) day and 16.5 (13-40) day, respectively. Acute GVHD occurred in 4 cases while chronic GVHD in 5 cases. The prospective OS for 3 years was (90.0±8.5)% after a median follow-up time of 17 months. Transplant related mortality was 1 case. Conclusion RIC-HSCT with FB+TBI is a feasible and effective alternative for MF patients.

Objective To investigate the role of micro-RNAs (miR-101 and miR-125a-5p) in autophagy of lipopolysaccharide (LPS) derived human THP-1 macrophages.Methods Human monocytic leukemia cell line THP-1 was cultured in vitro,and it was differentiated into macrophages after being induced with phorbol (50 μg/L) for 48 hours.THP-1 macrophages were stimulated with LPS in 0,250,500,1 000 μg/L respectively for 12 hours,miR-mimic was transfected into THP-1 macrophages as induced by Lipofectamine RNAiMAX,and the transfection efficiency of miRNA was determined with fluorescence microscopy.Enzyme linked immunosorbent assay (ELISA) was used to determine the levels of tumor necrosis factor-α (TNF-α) and monocyte chemotaxis protein-1 (MCP-1) in the supernatants of culture.Western Blot was used to detect the protein expressions of autophagy proteins ATG4D,Beclin1,and LC3 Ⅱ.The expression levels of miR-101 and miR-125a-5p were determined by quantitative reverse transcription-quantitative polymerase chain reaction (RT-qPCR).Results ① The releasing levels of TNF-α and MCP-1 induced by LPS with 250,500,1 000 μg/L were significantly increased as compared the cells without LPS stimulation [TNF-α (ng/L):1 336.1 ± 18.5,1 340.6±24.8,1 364.5± 14.9 vs.47.6±4.4;MCP-1 (ng/L):996.3 ±51.3,934.6±84.3,974.2±69.5 vs.21.3±6.5,all P ＜ 0.01],but no significant differences were found among the three LPS stimulation groups.The protein expressions of ATG4D,Beclin1 and LC3 1Ⅱ were up-regulated in the presence of different LPS concentrations (0,250,500,1 000 μg/L) for 12 hours in THP-1 macrophages (when compared with the cells without LPS stimulation,t value of ATG4D was 8.103,38.410,52.020,P value was 0.015,0.001,＜ 0.001;t value of Beclin1 was 3.026,5.328,3.482,P value was 0.047,0.034,0.037;t value of LC3 Ⅱ was 3.836,6.200,4.665,P value was 0.018,0.003,0.010),and the optimal concentration was 500 tg/L LPS.When THP-1 macrophages were stimulated with 500 μg/L LPS for 12 hours,the expression levels of miR-101 and miR-125a-5p were down-regulated significantly as compared with the cells without LPS stimulation [miR-101 (2-△ △Ct):0.68 ± 0.08 vs.1.95 ±0.26,t =8.047,P =0.001;miR-125a-5p (2-△ △Ct):0.23 ± 0.06 vs.1.69± 0.42,t =5.975,P =0.004].② The higher transfection efficiency was showed under fluorescence microscope.Westem Blot results showed the protein expressions of ATG4D,Beclin1 and LC3 Ⅱ were down-regulated as induced by an over-expression of miR-101 or miR-125a-5p in THP-1 macrophages,and more obviously down regulated by co-transfected with miR-101 and miR-125a-5p (compared with negative control group,t value was 14.550,5.855,14.180,P value was 0.005,0.014,＜ 0.001).Conclusion miR-101 and miR-125a-5p can inhibit the autophagy in LPS challenged THP-1 macrophages,and the potential mechanism might be related to target regulation of ATG4D.

Objective To examine the distribution of bacterial species and antimicrobial susceptibility profile of pathogens in febrile neutropenic patients.Methods A total of 355 bacterial strains were isolated from febrile neutropenic patients in Shanghai General Hospital from January 2005 to December 2012. Antimicrobial susceptibility testing was done by Kirby-Bauer method. The susceptibility testing results were analyzed according to CLSI 2014 breakpoints.Results Gram-negative bacteria accounted for 70.4% of the 355 isolates, while gram-positive organisms accounted for 29.6%. The most common bacterial species werePseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, Stenotrophomonas maltophiliaand Staphylococcus haemolyticus. Non-fermentative bacteria accounted for 53.2% of all the gram-negative bacterial isolates. All theEnterococcus and
Staphylococcus isolates were susceptible to linezolid, vancomycin and teicoplanin. All theStaphylococcus strains were resistant to methicillin.P. aeruginosa isolates were relatively more susceptible to cefoperazone-sulbactam, piperacillin-tazobactam and cefepime (>70%) than imipenem (40.8%) and meropenem (59.2%). All theK. pneumoniae isolates were susceptible to imipenem and meropenem and more than 70% of the isolates were susceptible to cefoperazone-sulbactam, amikacin. More than 80% of theA. baumannii isolates were susceptible to carbapenems, cefoperazone-sulbactam, amikacin, ciprolfoxacin and aminoglycosides. All the E. coli isolates were susceptible to carbapenems and more than 70% were susceptible to cefoperazone-sulbactam and ceftazidime. More than 90% of theS. maltophilia strains were sensitive to levolfoxacin, minocycline, cefoperazone-sulbactam and trimethoprim-sulfamethoxazole.Conclusions Our data suggest that gram-negative bacteria, especiallyEnterobacteriaceae and non-fermentative bacteria, are still the primary pathogens in febrile neutropenic patients. Antimicrobial resistant strains are prevalent. Such data of bacterial species and antimicrobial susceptibility proifle of pathogens in febrile neutropenic patients are useful for empirical antimicrobial therapy of such infections.