Objective ToanalyzethedistributionanddrugresistancesituationofKlebsiellapneumoniae(KPN)isolatedfromour hospital during 2014 .Methods The drug susceptibility test was carried out by the Siemens fully automatic bacterial identification /drug susceptibility test analyzer .The drug susceptibility test results were judged according to the (CLSI) M100‐S22 document (2012) .Results A total of 291 strains of KPN were detected from 8 486 submitted specimens ,in which 20 strains were multi‐drug resistant Klebsiella pneumoniae(MDR‐KPN)isolates were 213 strains ,48 strains were extended‐spectrum beta‐lactamases producing Klebsiella pneumoniae (ESBLs‐KPN) and 10 strains were carbapenem‐resistant Klebsiella pneumoniae(CR‐KPN) .The KPN detec‐tion rate had statistical difference among different specimens (P= 0 .000) .The constituent ratio of KPN isolation in the sputum specimen was highest(P=0 .000) .The KPN detection rate had statistical difference among different departments (P=0 .000) ,the constituent ratio of KPN isolation in the respiration department was highest (P=0 .000) .The resistance and sensitivity of KPN to different antibacterial drugs had statistical difference(P=0 .000) ,carbapenems had the lowest resistance rate .Conclusion KPN has relatively low resistance rate to commonly used antibacterial drugs ,its multi‐drug resistant strains are in the higher level ,which should arouse clinic and hospital infection management department must to pay attention to and adopt corresponding supervision measures .

Objective To compare the clinical efficacy of different margins in nephron-sparing operation for patients with small renal cell carcinoma. Methods From September 2008 to April 2013, a total of 64 patients with local renal cell carcinoma (T1a period) and treated with nephron-sparing operation were selected, and the clinic data were analyzed. According to cutting edge size gotten from the surgery, the patients were divided in to A group (cutting edge 1-5 mm group, 30 cases) and B group (cutting edge 6-10 mm group, 34 cases). The operation condition and recurrence rate and survival rate of two groups were compared. Results The operative time in A group was significantly shorter than that in B group:(130.1 ± 24.0) min vs. (152.3 ± 28.0) min, P<0.05. The drainage time and the level of serum creatinine in two groups had no significant differences (P>0.05). The 3-year recurrence rate and 3-year survival rate in two groups had no significant differences (P>0.05). Conclusions The clinical efficacy of different margins in nephron-sparing operation for small renal cell carcinoma is similar. But 1- 5 mm cutting edge size nephron-sparing operation has less complications, and the recurrence rate and survival rate does not increase. It is worthy of spread .

Objective To investigate the changes in cerebral hemodynamics in different body positions in healthy volunteers.Methods Sixty right-handed healthy male volunteers,aged 22-26,height 167-178 cm,weighing 51-67 kg,were randomly divided into 4 groups ( n =15 each): 15 degrees head-down tilt group (group Ⅰ ),30 degrees head-down tilt group (group Ⅱ ),15 degrees head-up tilt group (group Ⅲ) and 30 degrees head-up tilt group (group Ⅳ ).Blood flow signals of right middle cerebral artery (MCA) and extracranial internal carotid artery (EICA) were detected by transcranial Doppler and systolic blood flow velocity (Vs),diastolic blood flow velocity (Vd ),mean blood flow velocity (Vm),pulsatility index (PI) and resistance index (RI) were recorded at supine position (baseline),immediately,10 and 30 min after body position change(T1-3 ).Lindegaard index was calculated.Results Compared with the baseline,in group [Ⅱ Lindegaard index was increased at T2,while Vs and Vm of MCA were decreased at T2,3,in group Ⅳ Vs of MCA and PI of EICA at T2,Vd and Vm of MCA at T2,3 were decreased ( P ＜ 0.05),while there was no significant difference in the variables mentioned above in the other two groups ( P ＞ 0.05 ).Conclusion In healthy adults,cerebral blood flow velocity decreases in 30 degrees headdown and head-up tilt positions,however there is no change in cerebral blood flow velocity in 15 degrees headdown and head-up tilt positions.

Objective To investigate the expression of pancreatic thioredoxin-1 (TRX-1) in rats with acute necrotizing pancreatitis (ANP) and the effect of pretreatment of melatonin on its expression. Methods Male Spraque-Dawley rats (n = 12) were randomly divided to ANP group, melatonin group, control group with 24 rats in each group. The rats in ANP group received three intraperitoneal injections of 25 ml/kg body weight 6% L-arginine at an interval of 1 h to induce ANP. The rats in melatonin group received intraperitoneal injections of 25 ml/kg body weight 6% melatonin 30 min before ANP induction; rats in ANP group and control group received intraperitoneal injections of same amount of saline. Rats were sacrificed at 6 h, 12 h and 24 h after ANP induction. The serum level of amylase was measured and the pathological evaluation of pancreatic tissues was performed. The concentrations of malondialdehyde (MDA) and myeloperoxidase (MPO) in pancreatic tissues were measured. The expressions of TRX-1 protein were detected by immunohistochemistry and the expressions of TRX-1 mRNA in pancreatic tissues were determined by RT-PCR.Results In ANP group, serum level of amylase, MDA, MPO, TRX-1 mRNA and TRX-1 protein in pancreatic tissues were (3 012 ±1 425) U/L, (4.13 ± 1. 85)nmol/mg prot,(7.45 ± 1.26)nmol/mg prot, 0.68 ±0. 18, 66.8 ±8. 1, while they were (1 835±499)U/L, (3.03 ±2.12) nmol/mg prot, (5. 32 ± 1.06) nmol/mg prot, 0.50±0.09, 80. 29 ±8. 14, respectively in melatonin group, the values in melatonin group were significantly lower thanthose in ANP group (P < 0.05). The peak value of TRX-1 mRNA and TRX-1 protwein expressions shifted from 12 h after ANP induction in ANP group to 6 h after ANP induction in melatonin group. Conclusions The expression of pancreatic TRX-1 protein and TRX-1 mRNA in rats with ANP was significantly increased. Melatonin pretreatment could promote pancreatic tissues to express TRX-1 protein and TRX-1 mRNA, and may be protective for pancreatic tissues damages.

BACKGROUND:Laser therapy is a non-invasive and painless treatment that is considered to be an effective method suitable for the treatment of osteoarthritis due to its simplicity and non-invasive nature.Currently,the mechanism of action of laser therapy is unclear and the results of studies on its clinical application are controversial. OBJECTIVE:To review and summarize the latest research progress of laser therapy on chondrocytes,animal experiments and clinical efficacy,and to explore the possible mechanism of laser-mediated multi-pathway biological effects,so as to provide a theoretical basis for further research on the laser treatment of osteoarthritis of the knee joint. METHODS:A literature search was performed in CNKI,WanFang Data,VIP and PubMed databases for relevant literature published from 2018 to 2023,with"laser therapy,low level laser therapy,high level laser therapy,photobiomodulation,knee osteoarthritis,chondrocytes"as the search terms in Chinese and English,respectively.Together with 14 articles searched manually,70 articles were finally included for review. RESULTS AND CONCLUSION:Laser therapy in the treatment of knee osteoarthritis is mainly categorized into two types:low-level laser therapy and high-level laser therapy.Differences in laser parameters and treatment protocols have a direct impact on laser efficacy.When appropriate parameters are used,low-level lasers show positive effects in cellular experiments,animal models,and clinical efficacy.High-level lasers have been less studied in the treatment of knee osteoarthritis,but some preliminary clinical studies have shown positive results.Cell experiments have shown that low-level laser promotes chondrocyte proliferation and cartilage matrix synthesis,thereby reducing inflammatory response.Animal experiments have shown that low-level laser can reduce the release of pro-inflammatory factors,promote cartilage matrix synthesis,inhibit matrix degradation,and effectively improve the repair process of cartilage tissue.Low-level laser is also able to reduce oxidative stress damage and relieve pain in knee osteoarthritis.In clinical trials,both low-and high-level laser can reduce patients'pain and improve functional activities.The combination of laser therapy and exercise therapy modalities may improve the therapeutic effect.Lasers may affect intracellular signaling and cellular functions through photobiological or thermodynamic effects.This provides direct evidence that laser promotes articular cartilage regeneration.

Objective:To study the risk factors of necrotizing enterocolitis (NEC) after surgery for intestinal atresia.Method:From August 2013 to June 2020, children with intestinal atresia receiving surgery in our hospital were retrospectively reviewed. The patients were assigned into NEC group and non-NEC group according to the occurrence of postoperative NEC. Demographic data and clinical characteristics were summarized and the risk factors for postoperative NEC were analyzed using Logistic regression analysis method.Result:A total of 96 infants were enrolled and NEC occurred in 13 patients (13.5%) after surgery for intestinal atresia. Compared with the non-NEC group, the NEC group were diagnosed of intestinal atresia [4.0(1.5,6.0)d vs. 1.4(0,2.0)d, P<0.001] and received surgery [4.8(2.0,7.0)d vs. 3.1(1.0,4.0)d, P=0.034] at later ages. The incidences of complex intestinal atresia [76.9%(10/13) vs. 44.6%(37/83), P=0.030] and blood transfusion [46.2%(6/13) vs. 13.3%(11/83), P=0.007] in the NEC group were higher than the non-NEC group. Logistic regression analysis showed that the age of initial diagnosis of intestinal atresia ( OR=3.346, 95% CI 1.493~7.500, P=0.003), complex intestinal atresia ( OR=9.052, 95% CI 1.119~73.209, P=0.039) and blood transfusion ( OR=6.835, 95% CI 1.399~33.380, P=0.018) were independent risk factors for postoperative NEC. Conclusion:Patients with delayed diagnosis of intestinal atresia, complex intestinal atresia and blood transfusion within 48 hours after surgery should be monitored for the occurrence of postoperative NEC.

Objective: To investigate the effects of macrophage colony-stimulating factor (M-CSF) on the polarization and infiltration of M2 macrophages and the invasion and metastasis of tumor cells in ovarian cancer microenvironment. Methods: A co-culture system consisting of ovarian cancer cells (A2780 and SKOV3) and THP-1 derived macrophages was established in vitro. The M-CSF levels in culture medium and M-CSF mRNA levels in cancer cells and macrophages were detected by ELISA and qRT-PCR, respectively. The proportion of CD68+CD163+ M2 macrophages (polarization cells) was determined by flow cytometry. The invasive and metastatic ability of A2780 and SKOV3 cells after co-culturing with M2 macrophages were analyzed using Transwell assay. The expression levels of M-CSF, CD68+, CD163+ and E-cad in paraffin sections of 52 patients with ovarian cancer and 18 patients with benign ovarian tumor were detected by the immunohistochemistry staining, and their correlations and the relationship between M-CSF and clinicopathological features of ovarian cancer patients were analyzed. Results: The M-CSF levels in culture medium of the co-culture group (A2780 and SKOV3 cells co-cultured with M2 macrophages) were significantly higher than that of A2780 and SKOV3 cells alone (t=14.315 and 12.338, P＜0.01). Fluorescence quantitative PCR results showed that the increased M-CSF originated from the secretion of co-cultured ovarian cancer cells (t=29.915 and 36.826, P＜0.01). The proportions of CD68+CD163+ M2 macrophages in the A2780 cells co-cultured with M2 macrophages group and SKOV3 cells co-cultured with M2 macrophages group were (6.14±0.50)% and (7.32±0.67)%, respectively, which were significantly higher than that in the M2 macrophages alone group ([1.82±0.34]%, t=12.289 and 12.711, P＜0.01). Transwell assay showed that the co-culture environment enhanced the invasion of A2780 and SKOV3 cells (24.00±4.81 vs 75.20±6.42, t=11.058; 18.40±2.31 vs 61.60±9.66, t=7.537, P＜0.01). The expression levels of M-CSF in ovarian cancer tissues were positively correlated with the number of CD68+ cells and CD163+ cells (r=0.690 and 0.596, P＜0.01), and negatively with the expression levels of E-cad (r=-0.566, P＜0.01). Moreover, the expression levels of M-CSF and the number of CD68+ cells and CD163+ cells in ovarian cancer tissues were significantly higher than that in benign ovarian tumor tissues, however, the expression levels of E-cad were on the contrary. The expression levels of M-CSF in ovarian cancer tissues were significantly correlated with tumor stage, differentiation and lymphatic node metastasis (χ2=6.240, 6.612 and 4.544, respectively, P＜0.05). Conclusion The increased expression of M-CSF in ovarian cancer microenvironment may induce the polarization and infiltration of CD68+CD163+ M2 macrophages, and then promote the invasion and metastasis of ovarian cancer cells.

Objective: To evaluate the clinical efficacy of percutaneous tibial nerve electrical stimulation in the treatment of chronic pelvic pain syndrome. Methods: The clinical data of 28 patients with chronic pelvic pain syndrome from January to November 2018 in Dalian Third People′s Hospital were retrospectively analyzed. The patients were treated with percutaneous tibial nerve stimulation. The number of urination in 24 h, number of nocturnal urination, urine volume per urination, quality of life (QOL) score and digital pain intensity score before treatment and after treatment were compared; the patients were followed up for 3 months, and the recurrence and adverse reaction were observed. Results: Compared with those before treatment, the number of urination in 24 h and number of nocturnal urination after treatment were significantly lower: (11.6 ± 6.4) times vs. (20.6 ± 7.8) times and (2.5 ± 1.2) times vs. (5.2 ± 2.6) times, and the urine volume per urination increased significantly: (181.2 ± 65.6) ml vs. (125.4 ± 58.2) ml, the QOL score and digital pain intensity score decreased significantly: (2.6 ± 1.4) scores vs. (5.1 ± 0.8) scores and (2.9 ± 1.3) scores vs. (6.9 ± 1.4) scores, and there were statistical differences (P<0.05). During the follow-up period, none of the patients had recurrent symptoms and obvious adverse reaction. Conclusions Percutaneous tibial nerve stimulation is an effective way to treat chronic pelvic pain syndrome.

OBJECTIVETo establish a novel method to determine specific type of amyloidosis through laser microdissection and mass spectrometry (LMD/MS) based proteomic analysis.

METHODSThere were 138 formalin-fixed and paraffin-embedded (FFPE) biopsy samples of patients who were diagnosed as systemic amyloidosis used in this study. For each case, a 10 μm section stained with congo-red and positive amyloid deposits were identified under fluorescent light, followed by micro-dissection and mass spectrometry analysis. The amyloidosis subtype was confirmed based on the most abundant amyloid protein.

RESULTSThe tissue types of 138 specimens were as following: subcutaneous abdominal fat accounted for 26%, tongue for 19%, gingiva for 11%, kidney for 9%, intestine for 9%, heart for 6% and others for 20%. Specific types of amyloid were accurately detected in 121 cases, including 106 (87.6%) amyloid light chain (AL) type, 7 (5.8%) amyloid trans-thy-retin (ATTR), 2 (1.7%) amyloidogenic protein A (AA), 2 (1.7%) amyloid heavy chain (AH)/AL+AH, 2 (1.7%) fibrinogen alpha chain (AFib), 1(0.8%) amyloid apolipoprotein A-type II (AApoA-II) and one (0.8%) amyloid lysozyme (ALys). Diagnosis of amyloidosis was excluded in 5 cases. The types of twelve cases were indeterminate by LMD/MS. On the whole, LMD/MS reached 91.3% accuracy rate in amyloid typing. Commonly involved organs (for example, heart, kidney and liver) turned out to be suitable sources of FFPE samples with typing success rate of almost 100%. In contrast, MS analysis was successful in only 83.3% of subcutaneous abdominal fat samples.

CONCLUSIONLMD/MS method provided a more direct technique for accurate typing of amyloidosis in a single procedure.