Protein kinase C (PKC) belongs to the family of phospholipid-dependent serine/threonine kinase. PKC signaling has been implicated in the development of multiple human diseases including the central nervous system dysfunctions and cardiovascular disorders. It has been demonstrated that PKC can stimulate viral activation through multiple mechanisms and phosphorylate the HIV-1 p17 gag,Nef,Vif and Rev proteins as well. The phosphorylation of these proteins may have significant effects on HIV replication. Members of the PKC family participate at many levels in diverse signal transduction pathways involved in HIV cycles. PKC inhibition may be considered a logical mechanism which inhibits HIV-1 replication. Agents that induce HIV-1 replication could be used in conjunction with HAART to decrease or eliminate the latent reservoirs by forcing viral expression. A better understanding of the role of PKC could in form the search of specific target for new anti-HIV therapies.

ObjectiveTo evaluate the indication, the operation pattern and therapeutic effect of the ano-saving surgery in low rectal carcinoma. MethodsAccording to anorectal finger-examination, IRUS,and CT, 94 out the 161 rectal cancer patients were selected to have ano-saving surgery from August 1993 to December 1994.Excreting function, 5-year survival rate and local recurrence rate of the various operation were compared. ResultsThe perfomed rate of ano-saving operation in low rectal cancer was 58.4%. Among them, low anastomosis was done in 6 cases, ultra-low anastomosis in 48 cases,Park′s operation in 25 cases,and Bacon operation in15 . The death rate was 2.1%(2 cases).Incidence of anastomotic leakage after the surgery was 3.2%(3 cases), and only 13 cases had anastomotic narrowing(13.8%) within 1 year. The successful rates of excreting function after the surgery were respectively as follows: low anastomosis 100%, ultra-low anastomosis 97.9%,Park′s surgery 88.0%,and Bacon surgery 53.3%. The 5 year survival rates and the local recurrence rates were respectively,as follows: low anastomosis 83.3% and 0; ultra-low anastomosis 79.2% and 4.2%; Park′s 64.0% and 12.0%; and Bacon 66.7% and 13.3%,respectively. ConclusionsUltra-low colo rectum anastomosis becomes the main operative pattern to preserve anal sphincter in low rectal cancer.There is no difference in the 5-year survival rate and the excreting function among low, ultra-low anastomosis and Park′s operation, but the low and ultra-low colo-rectum anastomosis were obviously better than that of Bacon and Miles operation.The local recurrence rates of low and ultra-low colon-rectum anastomosis are lower than that of Miles′.There is no difference in the 5-year survival rate and local recurrence rate between Park′s, Bacon and Miles operation.

Objective Few prospective data are currently available on acute gastrointestinal hemorrhage (AGIH) as a complication in acute kidney injury (AKI).The aim of the present study was to find out clinical characteristics,incidence,etiology,risk factors,and outcome of AGIH in patients with AKI.Methods We performed a prospective study on an inceptione cohprt of 512 patients admitted for AKI in our hospital.Data on clinical risk factors for bleeding,frequency of occurrence of AGIH,in-hospital mortality were collected,and independent predictors of AGIH were identified.Results A total of 53 patients had AGIH as a complication of AKI,and 45 were upper AGIH.Fifteen patients had clinically severe bleeding.Independent baseline predictors of AGIH were severity of illness,cardiac failure,mechanical ventilation,low platelet count,chronic hepatic disease,liever cirrhosis,severe AKI.Inhospital mortality was 52.8％ in patients with AGIH,and 22.2％ in the other patients.AGIH was significantly associated with an increase in hospital mortality.Conclusions AGIH are frequent complications of AKI.In this clinical condition,AGIH is more often due to upper gastrointestinal bleeding and is associated with a significantly increased risk of death.Both renal and extrarenal risk factors are related to the occurrence of AGIH.

It was recently shown that several new synthetic 2-alkylsulfanyl-6-benzyl-3, 4-dihydropyrimidin-4(3H)-one (S-DABO) derivatives demonstrated anti-HIV-1 activity. Three of the derivatives namely RZK-4, RZK-5 and RZK-6 were used in this study to explore their inhibitory effects on a variety of HIV strains. These compounds at a concentration of 200 microg mL(-1) almost completely inhibited the activity of recombinant HIV-1 reverse transcriptase. All of the three compounds reduced replication of HIV-1 laboratory-derived strains, low-passage clinical isolated strain, and the drug resistant strain. In particular RZK-6 showed potent activity against the HIV-1 drug resistant strain. In general, the antiviral activities are similar in magnitude to nevirapine (NVP), which is a non-nucleoside reverse transcriptase inhibitor approved by FDA. The therapeutic indexes of these compounds were remarkable, ranging from 3704 to 38462 indicating extremely low cytotoxicity. These results suggest that the three S-DABO derivatives in this study have good potential for further development in anti-HIV-1 therapy. It may be particularly useful to target at the non-nucleoside reverse transcriptase inhibitors resistant HIV-1 strain.

In the present research, two Chinese rhesus monkeys were inoculated intravenously with 5000 TCID50 of SIVmac239. The changes in the numbers of CD4+T lymphocyte in peripheral blood,plasma viral loads, proviral DNA and humoral antibodies against virus were periodically monitored during 121 days. At the early stage of infection, proviral DNA had been detected in PBMCs, and infectious SIVmac239 virus had been isolated from PBMCs. At the same period, the numbers of CD4+T lymphocytes were significantly decreased, and maintained at low level during the 121-day period of infection. Plasma viral loads reached the peak at week 2 post-inoculation and kept at a steady state subsequently. Moreover, antibodies against viral proteins were detected from plasma. All the results showed that the two Chinese rhesus monkeys had been infected with SIVmac239 successfully. This animal model can be applied for further AIDS researches.

Objective To analyze the possible reasons of failed surgical treatment of acetabular fractures. Methods Various methods were used for positive patient identification, including according to Matta's X-ray assessment and Merle d'Aubigne & Postel hip function score of clinical standards for classification of acetabular fracture reduction surgery were not satisfied or not carried out a reduction and fixation,the clinical evaluation of hip joint as a "bad", occurrence of femoral head subluxation or dislocation, femoral head necrosis and other serious complications. From February 2000 to February 2008, 22 patients including 14 males and 8 females with an average age of 38.6 years (range, 18-72 years) were considered as failed cases. Results 45.5% of these cases were posterior wall fractures which were not given any fixation, 27.3% of them were posterior column fractures which were not fixed, 13.6% of them whose reduction and fixation of anterior wall fractures were not satisfied, and 9.1% of them were anterior column fractures which needed fixation. One case should take open reduction and iternal fixation instead of THA. The rate of misdiagnosis and mistaken diagnosis were 90% if only X-ray evaluation was made and this rate decreased to 8.3% if computed tomography was taken. The rate of wrong selection of operative approach was 100% in 10 cases of misdiagnosis, and which was 58.3% in 12 cases of correct diagnosis. In the 5 patients with correct diagnosis and selection of operative approach, the reasons of failed surgical treatment were due to imperfect surgical skills in 3 cases, and inappropriate fixation patterns in 2 cases. Conclusion The causes of the failure of surgical treatment for acetabular fracture might include preoperative missed diagnosis and misdiagnose, inappropriate approach, and an unreasonable internal fixation with unskillful technique.

Aim To establish and optimize the VSVG/HIV-1NL4-3 Luc pseudovirus model for anti-HIV drugs screening. Methods The infectivity of VSVG/HIV-1 NL4-3 Luc in 4 different cell lines was investigated according to the method of the lucifer-ase activity analysis system of Promega company. 3 different ex-perimental settings were used to detect the activities of approved anti-HIV drugs to confirm the feasibility and effectiveness of the system. Finally, some potential compounds were screened for their anti-HIV activities, and their antiviral activities against the pseudovirus were compared with HIV-1ⅢB . Results The pseud-ovirus showed the strongest replication ability in CRFK cells, and a clear dose-effect relationship was found between the report gene expression level and the virus quantity. Comparing the EC50 of different positive inhibitors against VSVG/HIV-1 NL4-3 Luc on 3 kinds of experimental conditions, 3rd scheme is the best. Finally, the system was used to screen compounds, the EC50 s a-gainst pseudovirus were similar to those in HIV-1ⅢB . Conclusion An optimized VSVG/HIV-1 NL4-3 Luc anti-HIV screening sys-tem has been successfully developed.

Background and purpose:Researches had indicated that about over 85%of malignant tumors highly express telomerase activity. So telomerase has become one of the important methods in the research field of tumor diagnosis and treatment. Nowadays, several reports about malignant tumor which over expresses hTERT targeting imaging with radionuclide labeled hTERT ASON had been published. In these reports, high quality of pictures can hardly be acquired because of poor anatomical and spacial resolution in nuclear imaging itself. Accordingly, in this study, we developed a method of detecting human telomerase in vivo with magnetic resonance imaging (MRI) and evaluate its feasibility. Methods:Firstly, Uniformly phosphorothioate-modified human telomerase reverse transcriptase antisense oligonucleotide (hTERT ASON) was labeled with Gd3+ through the bifunctional chelator 1, 4, 7, 10-tetraazacyclododecane-N, N’, N’’, N’’’-tetraacetic acid (DOTA) and iv vitro experiments were performed to characterize the antisense probes (for biodistribution and cellular uptake, 99mTc-DOTA-ASON was used in stead of Gd-DOTA-ASON). Then Gd-DOTA-ASON was injected intraperitoneally in pulmonary adenocarcinoma A375 nude mice tumor-bearing BALB/c for in vivo imaging using 7.0 T Micro MRI periodically, tumors and their surrounding tissues were defined as region of interest (ROI) to calculate the signal to noise ratio (SNR) of tumor to muscle using Gd-DTPA as control. Finally, immunohistochemical analysis of telomerase activity of each xenograft was operated 2 days after imaging. Results:The binding efficiency of Gd-DOTA-ASON reached was as high as 65%(63.2±2.4, n=6). And it can maintain 61%in fresh human serum and normal saline at 37℃over 24 h;A375 cells showed an uptake of 8.5%when incubated with 99mTc-DOTA-ASON;In comparing with DOTA-ASON and Gd-DTPA, cells transected with Gd-DOTA-ASON had higher SI when performed MRI with T1WI. The hTERT-expressing xenografts were obviously enhanced by Gd-DOTA-ASON at 0.5-6 h after injection and the SNR can reach 2.37, whereas obvious enhancement only could be found within 2 h after injection of Gd-DTPA. Both labeled and non-labeled antisense probes can suppress the activity of telomerase of A375 cells either in vitro or in vitro. Conclusion:Our research offers proof that Gd-DOTA-ASON can be used as tumor specific targeting MR probe for diagnosing malignant tumors with high expression of telomerase.

Objective:To preliminarily evaluate the effect of tension stimulation on the biological activity of and expression of fibrosis marker genes in keloid fibroblasts (KD-Fbs) .Methods:Three patients who were diagnosed with keloids and received surgical treatment were collected from the Department of Dermatology, Tangdu Hospital, the Fourth Military Medical University from January to March 2017. Human KD-Fbs were isolated from resected keloid tissues, and subjected to primary culture. The third- to sixth-passage KD-Fbs were divided into tension group and control group to be cultured in the tension-based chamber and control chamber respectively, and subjected to tension stimulation and normal culture respectively. Cell counting kit-8 (CCK8) assay was performed to assess the proliferative activity of KD-Fbs after 1-, 2-, 3- and 4-day culture, and the scratch assay to evaluate the migratory ability of KD-Fbs after 1- and 2-day culture. After 48-hour treatment, real-time quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of fibrosis markers type Ⅰ collagen, fibronectin and α-smooth muscle actin (α-SMA) in KD-Fbs respectively. Two-independent-sample t test was used for comparisons between 2 groups. Results:CCK8 assay showed that the proliferative activity of KD-Fbs was significantly higher in the tension group than in the control group after 1-, 2-, 3- and 4-day culture ( t=3.05, 7.00, 16.65, 15.19, respectively, all P< 0.05) . After 1- and 2-day culture, the scratch assay showed that the migration rate of KD-Fbs was significantly higher in the tension group (48.65%±3.96%, 100.00%, respectively) than in the control group (9.36%±1.14%, 50.35%±4.23%, t=16.53, 20.35, respectively, both P< 0.01) . Real-time quantitative PCR showed that the mRNA expression of type Ⅰ collagen, fibronectin and α-SMA was significantly higher in the tension group (3.04±0.20, 2.16±0.10, 3.76±0.24, respectively) than in the control group (1.00; t=17.57, 21.01, 20.25, respectively, all P< 0.01) . As Western blot analysis revealed, changes in the protein expression of the 3 fibrosis markers were consistent with their mRNA expression changes (all P< 0.05) . Conclusion:Tension may participate in the fibrosis in keloids by promoting the expression of fibrosis marker genes, and enhancing the proliferative and migratory ability of KD-Fbs.

Brain-derived neurotrophic factor (BDNF) exerts its survival-promoting effects on photoreceptors and retinal ganglion cells, however, delivery systems with little-to-no side effect are needed to sustain its controlled release and long-term efficacy. Our previous studies demonstrated that adipose-derived stem cells (ADSCs) are ideal delivery systems for gene therapy; moreover, ADSCs present unique properties like migration to damaged tissue sites, immunomodulation and anti-inflammation. Herein, we propose to employ ADSCs as the BDNF gene delivery vehicle. Different Analyses like flow cytometry, differentiation and cell proliferation assays etc demonstrated that BDNF were successfully transduced into ADSCs and the stemness of ADSCs was maintained even with the transduction. Real Time PCR and Western blot were used to measure mRNA and protein expressions of the BDNF-transduced ADSCs. The results demonstrated that the BDNF expression level of the lentiviral-BDNF transduced ADSCs is significantly increased and, moreover, enhanced the expression of other neurotrophic and downstream signaling factors. The data suggest that ADSCs are a reliable delivery vehicle for BDNF and could be used for the treatment of various diseases.