Objective To analyze and compare the clinical application values of procalcitonin (PCT),leukocyte count (WBC) and C-reactive protein(CRP) in elder patients with infection.Methods In patients(age≥ 65 yrs,axillary temperature ＞38.0℃)with infection or suspected infection,PCT,WBC,CRP and other bacteriological examination were performed.The electronic medical records from the HIS system of our hospital were analyzed retrospectively in combination with medical history.Results Of the enrolled 219 patients,65 ones were in control group,48 ones SIRS,51 ones sepsis and 55 ones MODS.There was a positive correlation between the level of serum PCT and the infection degree.The Spearman correlation coefficient was 0.706 (95％CI:0.616-0.797,P=0.000).Based on the highest Youden index (sensitivity+specificity-1),the best cutoff point of diagnosis for PCT was ＞0.341 μg/L (sensitivity 84.5％,specificity 55.8％),a analysis of receiver operating characteristic(ROC) curve about PCT,WBC and CRP was carried.Area under the curve (AUC) of PCT to controlled infection was 0.916 (95％CI:0.864-0.967,P=0.000).Based on the highest Youden index (sensitivity+ specificity-1),the judging threshold of PCT to infection controlled or not was 0.73 μg/L (sensitivity 84.6％,specificity 88.0％).PCT level after treatment ＞0.73 μg/L showed the uncontrolled infection,＜ 0.73 μg/L controlled.Conclusions PCT has a higher specificity for elder patients with infection.The variation of PCT level can guide the application of antibiotics,avoid abuse and decrease the occurrence of drug-resistant bacteria.

Objective To explore the best ratio of platelet-rich plasma (PRP) to activator and the synergistic action of thrombin and the growth factors in PRP gel on proliferation of human marrow-derived mesenchymal stem cells (hBMSCs). Methods The activator was made up of 1000 U bovine thrombin and 1 mL 10% calcium chloride solution. PRP was mixed with the activator at the ratios of 1:1, 5:1, 10:1, 20:1, 40:1 respectively in groups A, B, C, D and E. A quantitative sandwich enzyme-linked immunosorbent assay (ELISA) was used for examining the amounts of transforming growth factor-β1 (TGF-β1) and platelet derived growth factnr-AB (PDGF-AB) in PRP gel in each group after incubation for 0,1,8,24, and 120 hours. MTT assay was used to evaluate the effect of PRP gel in each group on hBMSCs proliferation. Results When PRP was mixed with thrombin, concentrations of PDGF-AB and TGF-β1 in PRP gel increased immediately and reached the peak value in 1 hour. The PRP gel in groups B, C, and D contained the highest amounts of PDGF-AB and TGF-β1, and accelerated hBMSCs growth re-markably. The cells proliferation in group A was inhibited. Conclusions The effect of PRP on the hBMSCs proliferation depends on the concentrations of PDGF-AB and TGF-β1 in PRP. Thrombin may influ-ence the hBMSCs proliferation by regulating concentrations of growth factors in PRP to certain extent, but too high a level of thrombin may inhibit hBMSCs growth.

Objective To explore the feasibility of labeling and tracing in vitro goat bone marrow mesenchymal stem cells (BMSCs) by bromodeoxyuridine (BrdU) on the basis of investigation of its optimal concentration, incubating time and cytotoxicity. Methods A healthy goat, aged 10 months old, male, weighing 32 kg, was used in this study. Bone marrow was aspirated. BMSCs were isolated and cultured using the adherence method in vitro. The fourth passage of BMSCs (P4) were incubated with BrdU at 5, 10, 15, 20 μmol/L as 5, 10, 15, 20 μmol/L BMSC groups. Cells were not labeled by BrdU as negative control. The following parameters were measured: induction, differentiation and determination of goat BMSCs; the optimal mass concentration and incubation time of 5-BrdU labeling; cell positive rate at 12, 24, 48 and 72 hours in each group using immunofluoreseenee; the cell survival rate after various concentrations of BrdU ladling by trypan-blue exclusion. Results The morphology of the primary and passage goat BMSCs was fusiform in shape. Goat BMSCs could differentiate into osteoblasts and chondrocytes following induction. BMSC nucleus showed green fluorescence under fluorescence microscope after being labeled by BrdU. The mean labeling rate increased with the increase in the concentration and incubation time of BrdU, and reached to (93.32± 3.25)% after incubation in 15 μmol/L, BrdU for 48 hours. There were no significant differences between 15 μmol/L BrdU for 72 hours, 20 μmol/L BrdU for 48 hours and 72 hours (P > 0.05), or between the other groups or time points (P < 0.05). The labeling rate of the blank control group was 0. The cell survival rate was all above 90% (P > 0.05). Conclusions BrdU can be used as a labeling marker for goat BMSCs. When the concentration is 15 μmol/L and the incubation time is 48 hours, the optimal labeling effect can be achieved. Goat BMSCs labeled with BrdU is of high efficiency and safety.

Objective To explore the feasibility of labeling in vitro rabbit osteoblasts with Qtracker and the features of Qtracker-labeled rabbit osteoblasts. Methods A healthy male rabbit, 3 months old, weighing 2 kg, was used in this study. After bone marrow was aspirated, bone marrow stromal cells (BMSCs) were isolated and cultured using the adherence method in vitro. The third passage of BMSCs was induced into osteablasts before incubation with Qtracker at concentrations of 1, 2, 4, 8, 16, 32 nmol/10~6 cells (Groups A, B, C, D, E, F respectively). Cells not labeled by Qtracker served as negative control (Group G). The following parameters were measured: induction, differentiation and determination of rabbit osteoblasts; the optimal mass concentration of Qtracker labeling by fluorescence microscopy and flow cytometry; the cell sur-vival rates at various concentrations of Qtraeker labeling by trypan-blue exclusion; Qtracker-labeled cell pro-liferation by MTr. Results The primary and the passage rabbit BMSCs were chiefly of fusiform shape. Rabbit BMSCs differentiated into osteoblasts following induction. The osteoblasts cytoplasm showed green fluorescence under fluorescence microscopy after being labeled by Qtracker. The mean labeling rate increased with the increased concentration of Qtracker, reaching up to (93.58±2.08) % after incubation at 8 nmol/ 10~6 cells by fluorescence microscopy, and (95.24±1.31) % by flow cytometry. There were no significant differences between Groups D, E, F(P>0.05), but significant differences were found between Groups A, B, C and Groups D, E, F (P<0.05). The labeling rate for Group G was 0. The cell survival rates were all above 96% (P>0.05) . No significant differences were found in the cell proliferation among various con-centrations (P>0.05). Conclusions Qtraeker can be used as a labeling marker for rabbit osteoblasts. When the concentration is at 8 nmol/10~6 cells, optimal labeling effect can be achieved. Rabbit osteoblasts labeled with Qtracker are of high efficiency and safety.

OBJECTIVETo observe the effect of traditional Chinese medicine (TCM) treatment according to syndrome differentiation on acute radio-reaction (ARR) in nasopharyngeal carcinoma (NPC) patients.

METHODSOne hundred and ninety-five NPC patients who received radiotherapy (RT) for the first time were randomly assigned to two groups: the control group (89 cases) was treated by RT alone for 7 weeks and the treatment group (106 cases) was treated by RT combined with oral taking TCM from starting of RT till 5 weeks after RT. The overall changes in total ARR score and ARR in different locations were observed weekly and compared.

RESULTSThe total ARR score in the treatment group was significantly lower than that in the control group (P<0.05). And the ARR scores of different organs, including skin, oropharyngeal mucosa, salivary glands, larynx, car, upper digestive tract, and central nervous system, in the treatment group were all lower than those of the corresponding organs in the control group. In addition, the ARR scores in both groups showed an ascending trend in the first 7 weeks and a descending trend from the 8th to the 10th week after beginning RT.

CONCLUSIONTCM treatment could relieve the ARR in the NPC patients without any affection on the efficacy of RT.

Adult ; Carcinoma, Squamous Cell ; drug therapy ; radiotherapy ; Combined Modality Therapy ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Nasopharyngeal Neoplasms ; drug therapy ; radiotherapy ; Phytotherapy ; Radiation Injuries ; diagnosis ; prevention & control ; Syndrome ; Treatment Outcome

AIM: To evaluate the anatomic and visual outcomes of 25-gauge vitrectomies combined with air tamponade for the treatment of idiopathic macular hole (IMH).METHODS: Thirty eyes of 27 patients with IMH were included in this prospective interventional study.All patients underwent 25-gauge pars plana vitrectomy (PPV) combined with phacoemulsification and air tamponade.Best corrected visual acuity (BCVA, logMAR), perimetry and multifocal electroretinography (mfERG) were conducted before and after the operation.Anatomical changes were evaluated with optical coherence tomography (OCT).RESULTS: The macular holes closed successfully in 28 eyes after the primary vitrectomy.The mean BCVA improved from 0.72±0.22 logMAR preoperatively to 0.29±0.18 logMAR postoperatively (P<0.001).In the visual field of central 10°, the average mean deviation (MD) decreased from-3.59±1.83 dB preoperatively to-2.51±1.36 dB postoperatively (P<0.001) and the average pattern standard deviation (PSD) decreased from 1.86±0.68 dB preoperatively to 1.33±0.32 dB postoperatively (P=0.001).The retinal response densities of mfERG in the foveal and perifoveal area increased significantly, and implicit times of rings 4-6 prolonged significantly (P<0.05).The symptom duration and baseline N1 amplitude densities at ring 1 had a significant impact on postoperative BCVA (P<0.001, P=0.001, respectively).CONCLUSION: The 25-gauge PPV and air tamponade with 1 day prone positioning produce favorable anatomic and functional outcomes.

OBJECTIVETo study the effect of H2O2 on the morphological pattern,vitality,proliferation,cycle period of rabbit intervertebral disc nucleus pulposus cells.

METHODSTen New Zealand white rabbits (2 to 3 kg, female) were used for isolating nucleus pulposus cells under sterilized condition. The culture solution with 15% FBS and DMEM/F12 (1:1) was applied for cell cultivation. After 90% cell fusion, the first generation was obtain and stimulated by H2O2 with different concentrations of 0 micromol/L (control group), 130 micromol/L,216 p.mol/L,360 Ipmol/L, 600 micromol/L,and 1000 micromol/L.

RESULTSCompared with the control group, there was little difference of the biological property (P>0.05) in 130 micromol/L and 216 micromol/L H202-treated groups. When the concentration of H2O2 attained 360 micromol/L, 600 micromol/L, and 1 000 micromol/L, the cells suffered aging,with increased cell vacuoles,decreased proliferation,and aging:related increase of 13-galactosidase dyeing. The cell cycle of many nucleus pulposus cells was blocked in G1 stage other than entering S stage. With increasing H2O2 concentrations, the aging degree was increased.

CONCLUSIONA certain concentration of H202 could induce early aging of nucleus pulposus cells,resulting in biological abnormalities of these cells.

Animals ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cellular Senescence ; drug effects ; Female ; Hydrogen Peroxide ; pharmacology ; Intervertebral Disc ; cytology ; drug effects ; Oxidative Stress ; Rabbits

The initial presentation of non-alcoholic steatohepatitis (NASH) is hepatic steatosis. The dysfunction of lipid metabolism within hepatocytes caused by genetic factors, diet, and insulin resistance causes lipid accumulation. Lipotoxicity, oxidative stress, mitochondrial dysfunction, and endoplasmic reticulum stress would further contribute to hepatocyte injury and death, leading to inflammation and immune dysfunction in the liver. During the healing process, the accumulation of an excessive amount of fibrosis might occur while healing. During the development of NASH and liver fibrosis, the gut-liver axis, adipose-liver axis, and renin-angiotensin system (RAS) may be dysregulated and impaired. Translocation of bacteria or its end-products entering the liver could activate hepatocytes, Kupffer cells, and hepatic stellate cells, exacerbating hepatic steatosis, inflammation, and fibrosis. Bile acids regulate glucose and lipid metabolism through Farnesoid X receptors in the liver and intestine. Increased adipose tissue-derived non-esterified fatty acids would aggravate hepatic steatosis. Increased leptin also plays a role in hepatic fibrogenesis, and decreased adiponectin may contribute to hepatic insulin resistance. Moreover, dysregulation of peroxisome proliferator-activated receptors in the liver, adipose, and muscle tissues may impair lipid metabolism. In addition, the RAS may contribute to hepatic fatty acid metabolism, inflammation, and fibrosis. The treatment includes lifestyle modification, pharmacological therapy, and non-pharmacological therapy. Currently, weight reduction by lifestyle modification or surgery is the most effective therapy. However, vitamin E, pioglitazone, and obeticholic acid have also been suggested. In this review, we will introduce some new clinical trials and experimental therapies for the treatment of NASH and related fibrosis.

BACKGROUNDPlatelet-rich plasma (PRP) as a storage vehicle of growth factors has been successfully used in clinical applications, but in most cases the platelets were autologous. However, the large volume of blood withdrawn has detrimental effects on patients with anemia or poor general health. To overcome these limitations, this study was designed to separate the growth factors in homologous platelet-rich plasma.

METHODSThe gel chromatography with Superdex-75 column was applied to separate PRP supernatants into 4 major fractions. Then the four fractions were vacuumed freeze-dried and re-dissolved in phosphate buffered saline. Proteins concentrations in PRP and in four fractions were detected by bicinchoninic acid protein assay; platelet derived growth factor-AB (PDGF-AB) and transforming growth factor beta1 (TGF-beta1) levels were determined by sandwich enzyme-linked immunosorbent assays. The effects of fractions on the proliferation of human marrow-derived mesenchymal stem cells (MSCs) were determined by 3-(4, 5- dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay.

RESULTSPRP supernatants were separated into four major fractions by gel chromatography. The proteins recovery was 96.72%. Of the four fractions, fraction B contained the highest TGF-beta1 and PDGF-AB levels, and the highest proteins concentrations. Cell proliferation curves of MSC demonstrated that fraction B and C induced a remarkable increase of MTT values compared to the untreated culture (P < 0.05), and the effects of fraction B and C showed no significant difference compared to the PRP group (P > 0.05). Fraction A and D showed no significant difference to the negative control group (P > 0.05).

CONCLUSIONSThe growth factors in PRP supernatants could be preliminarily separated into four fractions by gel chromatography, and the freeze-drying fractions retained the biological activity of growth factors. The growth factors were mostly presented in fraction B and C, and they promoted cell proliferation effectively.

Bone Marrow Cells ; cytology ; Cell Proliferation ; drug effects ; Chromatography, Gel ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Platelet Count ; Platelet-Derived Growth Factor ; isolation & purification ; pharmacology ; Platelet-Rich Plasma ; chemistry ; Transforming Growth Factor beta1 ; isolation & purification ; pharmacology

Computer Graphics ; Computer-Assisted Instruction ; Forensic Pathology ; Humans ; Imaging, Three-Dimensional ; Pathology ; education ; Pathology, Clinical ; Specimen Handling ; methods