Objective: To investigate the expression of histone deacetyase4 (HDAC4) in human liver carcinoma cell line Bel-7402 and to explore the regulatory effects of HDAC4 on the proliferation and differentiation of Bel-7402. Methods: Carcinoma cells Bel-7402 was treated with different concentrations of sodium phenylbutyrate (SPB), an inhibitor of HDAC4. Expression of HDAC4 mRNA in Bel-7402 cells was analyzed by RT-PCR before and after SPB treatment. Reverse microscope was used to observe the morphological changes of Bel-7402 cells. MTT assay and flow cytometry were adopted to describe the proliferation and cell cycle of Bel-7402 cells. Expression of P27 protein was determined by immunohistochemical method. The statistical analysis was performed using one-way ANOVA and student t test. Results: SPB significantly decreased the expression of HDAC4 in Bel-7402(0.88?0.13) vs (0.12?0.04), P

Objective To identify the incidence and risk factors of gastrointestinal bleeding in patients supported with mechanical ventilation over 48 hours. Method A total of 127 ICU patients supported with mechanical ventilation for over 48 hours were enrolled from January 1, 2007 to December 31, 2008 for the retrospective study. Exclusion criteria included the history of gastrointestinal bleeding and ulcer, recent gastrointestinal surgery, brain death and active bleeding from nose or throat. Demographics of patients including age, diagnosis at admission, duration of ICU stay, duration of mechanical ventilation, pattern and parameters of ventilation, ICU mortality, A-PACHE II score, multiple organ dysfunction score, and the results of biochemical assays including renal, hepatic and coagulation functions were recorded. Risk factors of gastrointestinal bleeding were analyzed by using univariate analysis And multiple logistic-regression analysis. Results Of the 127 patients, the incidence of gastrointestinal bleeding was 41.7% . and among them 3.9% patients suffered from clinically significant bleeding. However, the independent risk factors of gastrointestinal bleeding were the peak inspiratory pressure > 30 cmH20 (RR = 3.73, 95% CI = 1.59-9.46), renal failure (RR = 1.16,95% CI = 1.02 - 2.32), PLT count <50× 109 L-1(RR = 2.67, 95% CI = 1.32 - 15.78) and prolonged APTT (RR = 4.58, 95%CI = 2.32 - 12.96). The good entetal nutrition had a beneficial effect to the avoidance of gastrointestinal bleeding ( RR = 0.30, 95% CI = 0. 13 - 0.67). Conclusions The incidence of gastrointestinal bleeding is high in patients supported with mechanical ventilation, and the bleeding usually occurs within the first 48 hours. High pressure ventilator setting, renal failure, decreased PLT count and prolonged APTT are significant risk factors of gastrointestinal bleeding. However, the good enteral nutrition is the independent protective factors.

Objective To evaluate the effects of fluid resuscitation and large-volume hemofiltration (HVHF) on the Alveolar-arterial oxygen exchange in patients with refractory septic shock. Method A total of 89 intensive care patients with refractory septic shock treated with fluid resuscitation and/or HVHF were enrolled between August 2006 and December 2009. All the patients were randomly divided into two groups. In group A, patients were treated with fluid resuscitation, n = 41 cases) and in group B, patients were treated with large-volume hemofiltration and fluid resuscitation, n =48). The O2 content of central venous blood(CcvO2), arterial oxygen content (CaO2), Alveolar-arterial oxygen pressure difference (P(A-a)DO2), the ratio of arterial oxygen pressure/alveolar oxygen pressure (PaO2/PAO2), respiratory index (RI) and oxygenation index (OI) were checked. The levels of oxygen exchange in two groups were detected by arterial blood gas analysis before treatnent, 24 hour, 72 hour and 7 days after treatment. The APACHE Ⅱ scores in patients with refractory septic shock were measured before and the 7th day after treatment with HVHF and/or fluid resuscitation respectively. Data were analyzed by using t -test and chi-square test to compare the differences and ratio between two groups and were expressed in mean ± standard deviation, and the analysis of variance was done with SPSS version 12.0 software. Results ① The differences in CcvO2 and CaO2 between two groups were[(0.60±0.24) vs. (0.72±-0.28), P ＜0.05 and (0.84±0.43) vs. (0.94±0.46), P ＜0.05]; and the oxygen extraction rates (O2ER) were significantly different between two groups [(28.7±2.4) vs. (21.7±3.4), P<0.01];② The levels of P(A-a)DO2、ratio of PaO2/PAO2、RI and OI in group B were reduced more significantly than in group A (P<0.05 or P<0.01);③The APACHE Ⅱ scores in both groups were gradually reduced after treatment for 7 days, and the APACHE Ⅱscore in group B on the 7th day of treatment were lower than that in group A[(17.2 ± 6.8) vs. (8.2 ± 3.8), P ＜ 0.01]. Conclusions Fluid resuscitation and HVHF could improve alveolar-arterial-oxygen exchange in patients with refractory septic shock, and at the same time decreased the APACHE Ⅱ scores, improving the survival rate of patients.

Objective To observe the clinical efficacy of ulinastatin(UT) conjoined to high flow continuous blood purification( CBP) in the critical patients with multiple organ dysfunction syndrome(MODS). To evaluate the therapeutic potential of UT and CBP in systemic inflammatory response syndrome (SIRS) , severe sepsis( SS) , acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). Method A total of 122 cases of emergency and critical patients with a score of more than 15 counted up from APACHE H (acute physiology and chronic health evaluation 11 ) were randomly divided into Ulinastatin treatment group (UT group, n = 35) .continuous blood pu-rification(CBP group, n = 31),UT plus CBP (combine group, n = 30) and routine treatment group (control group, n =26). Routine treatment was given to patients of all groups, and patients of UT group had Ulinastatin 0.4 MIU given intravenously every 8 hours for 7 days in addition. Patients of CBP group were managed with continuous blood purification round the clock for 7 days and those of combine group were treated with UT plus CBP for 7 days.The efficacy of the treatment in four groups was assessed,and serum high sensivity reactive protein(hs-CRP) and IL-6 levels were measured on admission and comparison was made between values of biomarkers taken before and 1 d,3 d,and 7 d after treatment in four groups. The changes in WBCs,arterial gas analysis and the oxygena-tion index PaO2/FiO2 were checked, and at the same time, the APACHE II values and the incidence of MODS were compared within four groups. Results (1)One, three and seven days after treatment the plasma hs-CRP and IL-6 levels in UT and CBP groups were reduced significantly more than those in control group ( P < 0. 05), and in combine groups those were more dramatically lowered ( P < 0.05, P < 0.01). Before treatment there was no significance diffience in those values between groups, and there was on diffience in those values between 3 rd day and 7 th day after treatment ( P > 0.05). (2) The 1 st,3 rd and 7 th day after treatment the arterial gas PaO2/FiO2 index in UT and CBP groups was improved more than that in control group ( P < 0.05) , and it in combine group was most significant improved (P < 0.05,P < 0.01). The ALT and creatinine were lower than those in control group ( P < 0.05), and there were no significant differences in ALT and creatinine between groups before treatment (P > 0.05). (3) The 1 st,3 rd and 7th day afer treatment,the APACHE II values in UT and CBP groups were decreased more than those in control group ( P < 0. 05) , and therefore, the incidence of MODS was lower ( P < 0.05). Conclusions Ulinastatin could significantly inhibit the production of inflammatory cytokines and CBP could effectively eliminate inflammatory factors from blood, and the combination of these two approaches produce a more effective therapeutic potential for preventing MODS development.

In order to meet the needs of national clinical medical education strategy, to train medical students’ability of analyzing and solving problems, it is imminent to carry out the reform on comprehensive experiment in basic medi-cal teaching.During the practice, the basic medical college of Hebei North University tried to integrate functional ex-periment and morphological experiment, and had some experience to share with medical education colleagues.

Objective To study the therapeutic effects of omeprazoie in high-dose given by continuous intravenous infusion in the treatment of stress-related mucosal injury of G-I tract in intensive care patients.Method Totally 98 intensive care patients with stress-related mucosal injury(SRMI)were enrolled from August 2006 to October 2008 Department in Intensive Care Unit(ICU)of the Provincial Hospital Affiliated to Shandong University.All the patients were randomly divided into high-dose omeprazoie group(group A)and control group(group B).In group A,omeprazoie was administrated in loading dose of 80 mg Ⅰ.Ⅴ.in 5 minutes followed by maintenance dose of 8.0 mg/h intravenous infusion for 72 hours,while in group B,omeprazoie was given in dose of 40mg/8h intravenous infusion for 72 hours.The pH value of gastric juice was determined by German Roche pH test paper every 2 to 8 hours in the patients of both groups.The coffee like or red juice in the gastrointestine decompressor was observed.At the same time,hemoglobin(HB)was detected by Automatic blood cell analyzer Sysmex XE-2100,blood urea nitrogen(BUN)was determined by Automatic Analyzer Au5400,and buffer excess(BE)was checked by GEM Premie arterial blood gas analyzer in all patients.Data were expressed as mean ± standard deviation(x-± s)and the analysis of variance was done with SPSS 12.0 software.Comparison of mean value between two groups was conducted with t-test and the ratio was calculated by using chi-square test(X2 test).The change was considered as statistically significant if P value was less than 0.05.Results Four,eight,and 24 hours after treatment with omeprazole,the pH values in patients of group A were higher than those in patients of group B(four hous:6.63 ±0.62 vs.3.14 ±0.26,P＜0.01;eight hours and 24 hours:P＜0.05 or P＜0.01).At 8 hours and 24 hours after treatment,the HB was higher,BUN and BE were lower in group A than those in group B(P＜0.03 or P＜0.01).The total rate of hemostasis of upper G-I tract bleeding in group A was higher than that in group B(95.35%vs.78.19%,P＜0.05).Conclusions For treating the intensive care patients with SRMI,the continues intravenous infusion of omeprazole inhigh dose is superior to conventional dosage.

Objective To analyze oncological outcomes of 125I permanent implant brachytherapy (125I-PIB) in clinically localized prostate cancer. Methods Between June 2008 and June 2015, 121 patients with clinically localized prostate cancer were treated with 125I-PIB, and their average age was 72.37 years old. Before treatment, the average prostate-specific antigen (PSA) level was 17.7 ng/ml, prostate volume was (51.4 ±15.0) cm3, the International Prognostic Scoring System (IPSS) score was (21.6 ±2.4) scores. The intermediate-risk and high-risk patients were treated with adjuvant endocrine or supplemental external beam radiotherapy. All patients were followed-up. Biochemical recurrence was defined as the PSA nadir plus 2.0 ng/ml. Results Average follow-up time of the 121 patients was 41.81 months. The prostate volume was (23.1 ± 10.2) cm3, and the IPSS score was (9.7±3.3) scores. Rates of 5-year overall survival, biochemical recurrence-free survival and cancer-specific survival were 86.7 %, 76.7 % and 96.5 %, respectively. 5-year biochemical recurrence-free survival rates were 88.5 %, 67.5 % and 65.2 % in the low-, intermediate-, and high-risk groups, respectively. Biochemical recurrence-free survival did not differ significantly by three risk groups (P=0.103), but the difference between high-risk and low-risk groups was statistically significant (P=0.028). According to multivariate analysis, higher prostate-specific antigen (P=0.021), higher Gleason score (P=0.023) and higher clinical T stage (P=0.037) were the significant covariates associated with biochemical recurrence-free survival. The addition of hormonal therapy or external beam radiation therapy was associated with significantly better outcomes than brachytherapy monotherapy (P=0.036, P=0.027). As for complications, there were 4 cases of surgery (trans-urethral resection of the prostate) and 4 cases of cardiovascular complications. Conclusions The 125I-PIB can bring excellent oncological outcomes and acceptable complications in patients. Adjuvant endocrine or external beam radiotherapy for the intermediate-risk and high-risk patients may improve the outcome. Factors influencing efficacy include the high PSA, Gleason score and clinical T stage.

OBJECTIVETo develop an effective method for isolating and culturing single cancer stem cells.

METHODSThe capillary glass tube was stretched on fire and connected to a sterile plastic tube to prepare the single cell separation apparatus. Single SW480 cell clone spheres in serum-free culture were marked with CD133 and CK7, and the single cancer stem cells were separated and cultivated in 96-well plates or microdrop covered by paraffin.

RESULTSSW480 cell clone formation rate was about 1.04%, and the cell clone spheres highly expressed CD133 with low CK7 expression. The isolation of the single cancer stem cells showed a success rate of 98.99% using the separation device. The cell division profile was comparable between the cell cultures in microdrop and 96-well plates in the initial 2 cell divisions (P>0.05), whereas prolonged cell division occurred afterwards in the microdrop culture as compared to 96-well plate culture. The cell population expansion of the single cancer stem cells was similar between microdrop culture (11.5%, 22/192) and 96-well plate culture (9.2%, 17/184) (P>0.05).

CONCLUSIONSSingle SW480 cells can develop into cancer stem cell spheres. Microdrop culture is convenient and stable, and can be the primary choice for single cancer stem cell culture.

Background: Granular cell tumor (GCT) is an uncommon tumor, and gastrointestinal tract GCT is even more rare. Aims: To investigate the clinicopathological characteristics, treatment and prognosis of gastrointestinal tract GCT. Methods: Nine cases of gastrointestinal tract GCT from January 2017 to June 2021 at the 903rd Hospital of Joint Logistics Support Force, PLA and Jinhua Municipal Central Hospital Medical Group were retrieved. The clinical data, histopathological characteristics, treatment, and prognosis were retrospectively analyzed. Results: In the 9 patients with gastrointestinal tract GCT, ratio of male to female was 2:1, age at diagnosis was 19-60 years, with a median age of 52 years. Six GCT were found in esophagus, 2 in colorectum and 1 in anus. Endoscopic results showed submucosal protrusion or sessile polyps ranging in size from 2-12 mm with a median of 5 mm. Histology results showed that tumors were located in mucosa and/or submucosa, arranged in solid sheets or nests, with an infiltrative margin and inflammatory infiltrates. Tumor cells were mainly plump and polygonal with abundant cytoplasm and eosinophilic granules. Nuclei were small, the nuclear-cytoplasmic ratio was very low. Mitotic figure was rare. Immunohistochemistry results showed that S100 and CD68 proteins were positive in all patients, SOX10, CD56, Calretinin and Syn were positive in some patients, and CKp, Desmin, SMA, CD117, CD34, Dog1, and α-inhibin were negative in all patients. Esophageal and colorectal GCT patients received endoscopic mucosal resection (EMR) or endoscopic submucosal dissection (ESD). The anal GCT patient underwent local resection. Recurrence or metastasis were not observed during 9-53 months of follow-up. Conclusions: Gastrointestinal tract GCT is rare with non-specific clinical symptoms and submucosal protrusion or sessile polyps under endoscopy. Gastrointestinal tract GCT has special pathomorphology and immunophenotype. EMR or ESD is recommended for small and superficial lesions. Long-term follow-up should be performed.

The rise of nanotechnology has opened new horizons for cancer immunotherapy. However, most nanovaccines fabricated with nanomaterials suffer from carrier-related concerns, including low drug loading capacity, unpredictable metabolism, and potential systemic toxicity, which bring obstacles for their clinical translation. Herein, we developed an antigen self-assembled nanovaccine, which was resulted from a simple acryloyl modification of the antigen to induce self-assembly. Furthermore, a dendritic cell targeting head mannose monomer and a mevalonate pathway inhibitor zoledronic acid (Zol) were integrated or absorbed onto the nanoparticles (denoted as MEAO-Z) to intensify the immune response. The synthesized nanovaccine with a diameter of around 70 nm showed successful lymph node transportation, high dendritic cell internalization, promoted costimulatory molecule expression, and preferable antigen cross-presentation. In virtue of the above superiorities, MEAO-Z induced remarkably higher titers of serum antibody, stronger cytotoxic T lymphocyte immune responses and IFN-γ secretion than free antigen and adjuvants. In vivo, MEAO-Z significantly suppressed EG7-OVA tumor growth and prolonged the survival time of tumor-bearing mice. These results indicated the translation promise of our self-assembled nanovaccine for immune potentiation and cancer immunotherapy.