Objective To explore the application value of laparoscopic surgery in type 2 diabetes patients with acute cholecystitis.Methods 50 type 2 diabetic patients with acute cholecystitis were chosen as the research objects,the patients were divided into the observation group and control group according to the method of treatment, 25 cases in each group.Strict control of blood glucose levels in patients before operation,the observation group treated by laparoscopy,the control group were treated with conventional open surgery.Recorded the operation time and hemor-rhage,postoperative abdominal drainage,anal exhaust time,patient ambulation time and hospitalization time and other indicators.The patients were followed up to record the postoperative infection,occurrence of complications such as postoperative inflammation.Results There was no statistically significant differences between the two groups in oper-ation time(t =1.02,P >0.05).In the observation group,the intraoperative blood loss was (41.2 ±8.3)mL,postop-erative drainage volume was (107.5 ±16.4)mL,which were significantly lower than those in the control group,and the anal exhaust time was (1.7 ±0.3)d,total hospitalization time was (7.1 ±1.7)d,which were significantly shorter than those of the control group,the differences were statistically significant(t =3.43,3.78,3.43,4.21,all P <0.05).The observation group the complications included wound infection 1 cases,the complication rate was 4.0%, while in the control group,the complications included bile leakage,reflux esophagitis,incision infection,fat liquefac-tion,pulmonary infection and incisional hernia,the overall complication rate was 24.0%,there was statistically signifi-cant difference in the complication rate between the two groups(χ2 =4.54,P <0.05).Conclusion The laparoscopic surgery in the treatment of type 2 diabetic patients with acute cholecystitis has exact effect,and the patients recovery time is short,the complication rate is low.

Objective To explore the relationship between K-ras gene and cholangiocarcinoma by detecting the K-ras gene mutation of bile duct tissues.Method We studied all the patients who presented to our hospital from June 2010 to June 2012 with stenosis of the bile duct.There were 17 cases of cholangiocarcinoma and 17 cases of benign stenosis.From the DNA extracted from the paraffin tissues,we used the HRM assay for K-ras gene mutation.Result We found that the HRM method and the DNA typing had exactly the same result for the DNA content which confirmed the effectiveness of the HRM assay.Of note,the K-ras mutation rate was found to be significantly higher in the cholangiocarcinoma cases (11/17) when compared with the benign cases (3/17).Conclusion The mutation of the K-ras gene was closely related to cholangiocarcinoma.Our results suggest a new way to diagnosis cholangiocarcinoma.

This study was aimed to screen main active site to reduce blood lipid from Xin-Mai (XM) capsule and establish HPLC fingerprint of the site, in order to study the correlativity between active site and relevant fractions of its herbs. Solvent extraction was used to separate XM capsule into different polar fractions. Intraperitoneal injection of 75% egg-yolk emulsion was used to establish mice hyperlipidemia models. And the active site was screened. Chromatographic fingerprints of the site and relevant fractions of its herbs were configured by HPLC analysis. The
retention time of peaks was utilized as index to evaluate the correlativity. The results showed that lipid-lowering effect of ethyl acetate extract and garlic essential oil was significant (P<0.01). Fingerprint of the active site in XM capsule was established with 28 fingerprint peaks and the assignment results of 27 peaks were indicated. It was concluded that the active sites to reduce blood lipid of XM capsule were ethyl acetate extract and garlic essential oil. The established fingerprint method can effectively determine the correlativity between the active site and its relevant fractions, which contributed to pharmacodynamic material foundation and quality standard.

This study was aimed to develop an HPLC method for the determination of hesperidin,magnolol,honoki-oland liquiritin in Soft Capsule Jia-Wei Huo-Xiang Zheng-Qi (JWHXZQ). AKromasil C18 column (250 mmí4.6 mm, 5 μm) was used with water-methanol as mobile phasegradient elution. The flow rate was 1.0 mL·min-1, and the de-tecting wavelength was at 287 nm. The results showed that the linearity ranges ofhesperidin,magnolol,honokioland liquiritinwere 4.47-178.70 μg·mL-1, 3.42-136.96 μg·mL-1, 3.49-139.48 μg·mL-1, 3.92-157.20 μg·mL-1, respec-tively (r>0.999). The average recoveries of them were 99.48%, 99.05%, 99.57% and 99.79%, respectively. It was concluded that the method was accurate, sensitive and specific for quality control of Soft Capsule JWHXZQ.

Objective To investigate the effect of chronotherapy (chemotherapy plus timely medication) on neutrophils in breast cancer patients with neoadjuvant chemotherapy.Methods Fifty breast cancer patients with neoadjuvant chemotherapy were selected from 2011 to 2014 and divided into chronotherapy group and conventional treatment group (conventional group) by random number table method with 25 cases each.Conventional group received conventional neoadjuvant chemotherapy and conventional drug treatment.Chronotherapy group received neoadjuvant chemotherapy combined with chronochemotherapy and timely medication treatment,and applied clinical nursing care pathway.Both groups were conducted three cycles of chemotherapy,three weeks after the first and the second chemotherapy,the results of routine blood and liver function were compared in patients between two groups.The number of readmission and fever because of agranulocytosis were calculated.Results The two groups of patients were successfully completed three cycles of chemotherapy,after the first cycle of chemotherapy,the neutrophil was (4.40 ± 2.20)x109/L in chronotherapy group,and (3.18 ± 1.35) × 109/L in conventional group;after the second cycle of chemotherapy,the neutrophil was (3.95 ± 1.58) × 109/L,and (2.83 ± 1.49) x 109/L in conventional group,the two groups were statistically significant,t=2.375,2.563,P ＜0.05.Two cases needed readmission in chronotherapy group during chemotherapy,accounting for 8％(2/25),much lower than conventional group of 8 cases accounting for 32％(8/25).Conclusions The use of chronotherapy in breast cancer chemotherapy has less neutropenia,mild side effects of bone marrow suppression.It is an effective and safe viable option.

Abstract: To report the diagnosis, treatment and outcome of 4 patients with hematological diseases complicated with Aeromonas hydrophila bloodstream infection in the Second Affiliated Hospital of Kunming Medical University, further clarify the importance of blood culture and deepen the clinical understanding of the disease. Four patients with hematological diseases complicated with Aeromonas hydrophila bloodstream infection treated in the Second Affiliated Hospital of Kunming Medical University from 2017 to 2021 were recruited as the study objects. The clinical manifestations, blood culture collection, detection time of Aeromonas hydrophila, laboratory examination, treatment and prognosis of the patients were retrospectively analyzed. In this study, 4 cases were male patients with hematological diseases, who were in myelosuppression after chemotherapy. After fever, blood culture was collected and Aeromonas hydrophila was detected. The positive time of blood culture in 4 cases ranged from 4 to 11 hours. The results of antibiotic sensitivity showed that it was highly sensitive to the second, third and fourth generation cephalosporins, quinolones and carbapenems. Four patients were treated with imipenem cilastatin sodium in the early stage, and one patient recovered after active anti infection and leukocyte raising treatment. One patient did not complete chemotherapy due to a request for discharged, and the follow-up was unknown. Two patients developed rapidly into necrotizing fasciitis and died later. Hematological diseases complicated with Aeromonas hydrophila bloodstream infection are rare, but the mortality rate is high. For patients with repeated fever and considering infection, blood culture should be carried out as soon as possible to confirm the pathogen and drug sensitivity test. During clinical treatment, the treatment should be adjusted in time in combination with the patient's situation. In addition to anti-infection treatment, the patient's immunity should be improved and the development of necrotizing fasciitis should be vigilant. Keywords: Aeromonas hydrophila; hematologic diseases; leukemia; bloodstream infection; blood culture; necrotizing fasciitis

Abstract: Objective　To prepare a microparticle delivery system that regulates the release rate of extracellular vesicles (EVs), and to exert long-term enhancement of liver cell proliferation after only one intervention. Methods EVs was extracted by differential centrifugation. The structure of the EVs was observed by transmission electron microscopy and the membrane marker protein of EVs was detected by Western blotting. EVs-PLA microspheres with "core-shell" structure were prepared by emulsion-solvent evaporation method. Scanning and transmission electron microscopy were used to detect the morphology of EVs-PLA microspheres and EVs. The release test detected the release behavior of EVs in EVs-PLA microspheres. Scanning electron microscopy was used to detect the morphological changes of EVs-PLA microspheres at 8 weeks of release. EVs-PLA microspheres were co-cultured with hepatocytes, and Phalloidin/DAPI staining was used to observe the cell morphology and evaluate the cytotoxicity of the microspheres. CCK8-test was used to evaluate the cell proliferation activity. Western blot analysis was used to detect extracellular vesicles membrane marker protein expression. Results Comparing the ability of hepatocyte proliferation in the group treated with EVs-PLA microspheres and the control group, it was found that EVs-PLA microspheres did not cause cell apoptosis and mutation in cell structure, had biocompatibility and no cytotoxicity. The EVs-PLA microspheres with "core-shell" structure regulated the release behavior of EVs, which can continuously release EVs, exerting a continuous biological role in promoting hepatocyte proliferation after a single intervention. Conclusions The EVs-PLA microspheres can control-release EVs and promote hepatocyte proliferation continuously after a single intervention, providing a reference for further exploration of EVs-loaded delivery systems in promoting liver regeneration.

Objective:To study the differentiation capacity of the fibroblast-like cells isolated from human skin dermis into mesenchymal stem cells, and to explore the feasibility to use these cells as alternative cell source of autologus bone marrow mesenchymal stem cells (BMSCs ) for regeneration of tissue inj uries and defects. Methods:Full thickness skin samples were obtained from the abdomen of surgical patients, then digested with dispase and collagenase Ⅰ subsequently. Thereafter, the digested cells were collected and cultured, followed by suspension with serum free medium containing N2,B27,basic fibroblast growth factor (bFGF),and epidermal growth factor (EGF).The skin dermis derived spheroids (SDDSs)were collected and monolayer cultured in serum-containing medium.Finally,the cells were characterized by immunofluorescence staining and differentiation assays.Results:The dermis derived cells proliferated and formed SDDSs in the suspension of serum-free medium. After monolayer cultivation in serum-containing medium, the cells from spheroids were successfully expanded to large number. The cells expressed mesenchymal stem cells markers CD90, CD105 and vimentin. Under osteogenic,chondrogenic and adipogenic differentiation conditions,these cells were differentiated into the alizarin red,safranin O, and oil red O staining positive cells, displayed similar differentiation traits with BMSCs. However,safranin O staining was weaker in the dermis derived cells than BMSCs. Conclusion:A kind of fibroblast-like cells exist in human skin dermis, and have osteocytic, chondrogenic and adipogenic differentiation potentials,demonstrating that these cells will be utilized as a novel cell source for repairing the tissue injury and defect in clinic.

OBJECTIVETo investigate the molecular epidemiological characteristics and antibiotic resistance profiles of Vibrio cholerae O139 in Shandong province.

METHODSA total of 13 strains of V. cholerae O139 (9 clinical strains and 4 environmental strains) isolated from cholera epidemics in Shandong province since 1997 were recovered and confirmed with serum agglutination and biochemical reaction. Pulsed-field gel electrophoresis (PFGE) was carried out for molecular subtyping. Virulence genes and drug resistance related genes were detected by PCR. Antibiotic susceptibility tests were performed using micro-broth dilution method.

RESULTSThirteen strains of V. cholerae O139 were differentiated into seven pulsetypes. One clinical strain and two environmental strains isolated from Jining in 2013 were clustered into the pulsetype namely KZGN11O139. CN0077, and an identical PFGE pattern of KZGN11O139. CN0002 was found among three clinical strains from Jinan in 2005, Jining in 2005 and Heze in 2009. Other pulsotypes were unique in China and found only in Shandong province. Because of deletion of ctxAB and tcpI, the PFGE patterns of two strains isolated from Yantai in 2000 and 2004 were different from other 11 strains which harbored ctxAB, tcpA, tcpI, rtxA, hlyA and toxR. All strains contained one or more drug resistance related genes such as intI 1, intI 4 and sxt, and were resistant to two kinds of antibiotics at least. Among the 12 kinds of antibiotics, the resistant ratioes to kamamycin, trimethoprim-sulfamethoxazole, ampicillin and gentamicin were 11/13, 9/13, 7/13 and 7/13, respectively.

CONCLUSIONMolecular subtyping indicates possible epidemiological links among V.cholerae O139 in Shandong province, and almost all strains were toxigenic and drug resistant.

China ; Cholera ; Cholera Toxin ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Epidemics ; Humans ; Molecular Epidemiology ; Polymerase Chain Reaction ; Vibrio cholerae O139 ; Virulence

Chronic kidney disease (CKD) is becoming an alarming health burden worldwide, however, there is still lack of early biomarkers and effective treatment options. Thus, in the upcoming era of precision medicine, searching for the sensitive, non-invasive biomarkers has been the cornerstone and major challenge in the management of CKD. Urine contains rich biological information which could be an ideal source for non-invasive biomarkers of CKD. This review will discuss the recent advances in biomarker study from urine sediment, urine supernatant and urinary extracellular vesicles with special interest in gene transcript (miRNA, mRNA) biomarkers. Besides, the challenges and future directions for urinary gene transcript biomarker study will be discussed.
Biomarkers ; urine ; Humans ; MicroRNAs ; urine ; RNA, Messenger ; urine ; Renal Insufficiency, Chronic ; diagnosis ; urine