Objective To investigate the effects of Lycopene ( LP) on immune function of ovarian cancer rats′and its mechanism .Metho ds Sixty ovarian cancer rat models made by injecting ovarian cancer cell line NUTU-19 were divided into five groups randomly:model group,Cisplatin group(2 mg/kg)and LP(40,20,10 mg/kg)group,n =12;the LP and Cisplatin were given by intraperitoneal injection ,once every two days for 5 times.The rats general states and tumor growth conditions were observed ,the tumor tissue morphological changes was observed by H&E staining;the tumor weight ,inhibition rate,spleen lymphocyte transformation rate were de -tected;and the content of IL -2 and TNF-αin serum were detected .The present of CD4 +,CD8 +in blood were detected and the ratio of CD 4 +/CD8 +was calculated .Results The food situation of the rats in LP groups was improved,tumor was reduced,activity and hardness were poorer ,necrosis presented ,tumor cell shrinkage and oth-er pathological morphological changes appeared ,especially the rats in LP 40 mg/kg group .Compared with model group,the tumor weight in LP(40,20 mg/kg)groups was decreased,the inhibition rate and the spleen lymphocyte transformation rate were increased;the contents of IL-2 and TNF-αwere decreased;the CD8+present was de-creased,while the CD4 +present in LP 40 mg/kg group was increased,the ratio of CD4 +/CD8 +was increased. All of the difference above were significant (P<0.05,P<0.01).Conclusion LP can effectively improve the immune function of ovarian cancer rat;which perhaps relates to its effects of reducing the inflammatory cytokines levels and increasing the ratio of CD 4 +/CD8 +.

Objective To improve the awareness,diagnosis and treatment of pneumocystis carinii pneumonia (PCP) after renal transplantation.Methods A retrospective review was performed in 28 patients who underwent renal transplantation and developed PCP afterwards.The main clinical manifestations were fever(28 cases),nonproductive cough(28 cases),chest distress (12 cases).Occurrences of PCP were described 1.5 to 7 months after the renal transplantation.There were 10 patients treated with tacrolimus (FK506 2-6 rag/d,FK506 concentration 4-10 ng/ml) and 18 patients treated with cyclosporine (CsA 200-500 mg/d,CsA trough level:150-250 ng/ml) based immunosuppressive regimen.Anti-CD_(25)~+ monoclonal antibody (anti-CDCD_(25)~+mAb) was used in 10 cases for immune induction before operation while single steroid in 18 cases.Creatinine of patients with PCP was 70 to 106 μmol/L.CD_4~+ lymphocyte counts of the peripheral blood were 245±32/μl before PCP treatment and 536±25/μl after recovery.The most abnormal chest radiological findings were bilateral patchy ground-glass opacity.All the patients were diagnosed with PCP by bronchoalveolar lavage.Treatment was performed by reducing immunosuppressive agents and giving SMZco.Nineteen patients who had a PaP2 less than 70 mm Hg were given intravenous small-dose steroid.Results All the patients recovered from PCP 2 to 3 weeks after treatment.One patient experienced recurrence half year later.Five patients with higher creatinine after treatment recovered to normal levels after stopping the treatment of SMZco.No significant differences were seen in PCP patients treated with CsA and FK506,P＞0.05.The similar results were observed in use of anti-CDCD_(25)~+ mAb and single steroid,P＞0.05.Significant differences were observed in PCP patient peripheral blood CD_4~+ lymphocyte counts before and after treatment (P=0.001).Conclusions Patients who have fever,cough and hypoxia,chest imaging showing bilateral lung interstitial inflammation,might be PCP patients in the early post-renal transplantation period.Effective treatment should be performed by reducing immunosuppressive agents and giving SMZco.

Objective To design a new vermilion musculomucosal flap to reconstruct the normal appearance of unilateral cleft lip,and to analyze the clinical effects.Methods All patients were repaired with Millard method,and vermilion musculomucosal flap was formed with fixed point method to repair vermilion.Normal function and morphology were recovered by reconstructing lip beads and forming a peak of cupid's bow.Results 78 cases were enrolled in this study.They were followed-up for 6 to 36 months after operation.Except for 6 cases of nasal deformity and 5 mild lift of vermilion of affected side,67 cases obtained satisfied nasolabial form and function,with full decline and symmetry of cupid's bow peak,and well form of nasolabial fold,and natural transition of vermilion,and significance of lip beads,and inconspicuous scars.Conclusions Appling vermilion musculomucosal flap for reconstructing vermilion of unilateral cleft lip has good effect,and it is simple and feasible.It can be a regular surgery method,and is worthy of clinical promotion.

Objective To study the effects of oncolytic adenoviruses ZD55 harboring IL-24 gene (ZD55-IL-24) on the apoptosis of human melanoma cell line A375. Methods The oncolytie adenoviruses ZD55-IL-24 were verified by PCR. Then, the viruses were propagated, purified, and titrated by HEK293 cell plaque assay. A375 cells were cultured, divided into three groups transfected with ZD55-1L-24, ZD55 fused with enhanced green fluorescent protein (ZD55-EGFP), and replication-deficient adenovirus ZD55 carrying IL-24 gene (AD-IL-24), respectively. The multiplicity of infection was 0.1, 1, 10 and 100, respectively.Subsequently, the eytotoxity of these viruses and proliferation of A375 cells were determined by crystal violet staining and methyl thiazolyl tetrazolium (MTT) assay, respectively. The expressions of EIA and IL-24 protein were detected by Western blot in A375 cells. Results PCR verified that the adenoviruses ZD55-IL-24 contained IL-24 gene without wild adenovirns contamination. Crystal violet staining revealed that ZD55-IL-24 had an obvious eytotoxic effect on A375 cells, and MTT assay indicated that ZD55-IL-24 inhibited the proliferation of A375 cells in a time-and concentration-dependent manner. As shown by Western blot analysis, ZD55-1L-24 expressed IL-24 and E1A protein in A375 cells with a high efficiency. Conclusions The oncolytic adenoviruses ZD55-IL-24 can efficiently express IL-24 gene, inhibit the proliferation of, and induce the apoptosis in A375 cells.

Objective To evaluate the antitumor effect of oncolytic adenovirus expressing human IL-18 gene on malignant melanoma implanted in nude mice.Methods BALB/c nude mice were subcutaneously inoculated with A375 cells to establish a model of malignant melanoma.When the volume of implanted tumor reached 100-150 mm3,murine models were randomly divided into three groups to receive a 3-day intratumoral injection of IL-18 gene-expressing human oncolytic adenovirus named ZD55-IL-18,IL-18 gene-expressing adenovirus named Ad-IL-18,phosphate buffer saline(PBS),respectively.The tumor size was measured at an interval of 4 days for 9 weeks.Hematoxylin and eosin (HE)staining was performed to observe the morphological changes of tumor cells.The protein expression of IL-18 and E1A.microvessel density in tumor tissue,and apoptosis of tumor xenografts were detected by immuno fluorescence assay,immunohistochemistry and in situ end labeling technique (TUNEL).respectively.Results The treatment with ZD55-IL-18 significantly inhibited the growth of tumor.Forty-four days after the treatment,the mean tumor volume was 1039.378±29.67 mm3 in ZD55-IL-18-treated mice.significantly smaller than that in Ad-IL-18 treated mice(2900.46±62.65 mm3)and PBS-treated mice(3980.24±63.78 mm3).HE staining showed that the nuclei of tumor cells were heavily stained with few nucleoli in ZD55-IL-18-treated mice.Increased positivity rate of IL-18 was noticed in ZD55-IL-18-treated mice vs.AD55-IL-18-treated mice(83.4%±3.2%vs 24.4%±2.1%.P＜0.01).Moreover,immunofluorescence assay revealed the presence of E1A protein in tumor tissue.A decrease was found in the microvessel density in ZD55-IL-18-treated mice compared with the PBS-treated mice(P＜0.01).The apoptosis rate in tumor cells from high to low was 86.28%±3.25%in ZD55-IL-18-treated mice,43.67%±3.46%in Ad-IL-18-treated mice,and 10.73%±2.48%in PBS-treated mice;there was a significant difference between the three groups(all P＜0.05).Conclusion The oncolytic adenovirus expressing human IL-18 gene,ZD55-IL-18,has a significant inhibitory effect on the growth and metastasis of malignant melanoma implanted in nude mice.

The percutaneous transhepatic portal approach is the most commonly used technique for islet transplantation, largely owing to its safety and minimally invasive characteristic. Bleeding complications after islet transplantation are rare. A case of type 1 diabetes mellitus (T1DM) was treated in Tianjin First Center Hospital, who had a massive intra-abdominal hemorrhage after percutaneous transhepatic portal vein catheterization for islet transplantation. Through the review of the overall development of the case, we aim to improve the awareness of the complications of islet transplantation, to reduce the incidence of complications after percutaneous transhepatic portal vein transplantation, and to provide experience.

Objective To retrospectively compare the efficacy of Serva NB1 collagenase with Vitacyte GOLD collagenase on islet isolation of pancreas.Methods All the human pancreata were obtained from Chinese organ donors.In GMP laboratory,the pancreata were trimmed and distended with Serva NB1 collagenase (Serva NB1,n =12) or Vitacyte GOLD collagenase (Vitacyte GOLD,n =5) and digested according to a modified Ricordi semi-automatic protocol,and the digestion duration was recorded.The digested islets were then collected and washed,followed by the continuous density purification in a Cobe 2991 cell separator.The islet yield,purity,viability and glucose-stimulated insulin release (GSI) were determined each time after purification.Quantity and quality of isolated islets were determined by digestion efficacy.Results The digestion duration in Vitacyte GOLD collagenase group was significantly shorter than in Serva NB1 collagenase group to achieve the same digestion endpoint (P＜ 0.05).The islets yields of different sizes were variable between the two groups.The Vitacyte GOLD collagenase digestion produced more islets with a diameter range of 50-100 μm than the ServaNB1 collagenase digestion (P＜0.05),but the latter yielded more islets with a diameter range of 251-300 μm and 301-350μm (P＜0.05).There was no significant difference in total islets yields,viability,and GSI between two collagenase digestions (P＞0.05).Conclusion Both Vitacyte GOLD collagenase and Serva NB1 collagenase can be used for the clinical islet isolation in China.

Objective: To investigate the effect factors of liver enzymes elevation by monitoring the liver function changes before and after intraportal islet transplantation. Methods: 16 diabetic patients who received intraportal islet transplantation in our hospital were analyzed. The levels of aspartic aminotransferase (AST), alanine aminotransferase (ALT)and total bilirubin (TBil)were monitored after islet transplantation. Results: Among those 16 diabetic patients who received intraportal islet transplantation, 11 patients showed an increased AST and 8 patients showed an increased ALT, among which a 2.5-fold increase in AST was observed in 4 patients and over 1.5-fold elevation of ALT was observed in 3 patients. The level of TBil were in the normal range before and after transplantation in all patients. Transplanted tissue volume of islet was the main factor for significantly increased AST (P<0.05) in this study. It is also shown that the change in portal pressure is related to the AST elevation after islet transplantation. Conclusions The amount of transplanted islet tissue volume is related to the liver enzymes elevation after intraportal islet transplantation. Therefore, the improvement of the purity of islet to reduce the amount of transplanted tissue could be benefit to prevent the liver injury after islet transplantation. Meanwhile, the purified islets should be injected as slowly as possible to maintain a stable portal pressure.

Objective: To explore mechanism of lung injury of rats induced by inhalation of white smoke from burning smoke pot. Methods: Forty-eight Sprague Dawley rats were divided into control group (n=12) and injury group (n=36) according to the random number table. Rats in injury group were placed in smoke-induced injury experimental equipment fulled with white smoke from burning smoke pot for 5 minutes to make lung injury, and rats in control group were placed in smoke-induced injury experimental equipment fulled with air for 5 minutes to make sham injury. Six rats in injury group at post injury hour (PIH) 6, 24, and 72 and six rats in control group at PIH 72 were collected to observe pathological changes of lung tissue and pathological score of rats in the two groups by hematoxylin-eosin staining, to detect expression of nuclear factor-κB (NF-κB) p65 mRNA in lung tissue of rats by reverse transcriptional polymerase chain reaction, and to detect content of tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6 in lung tissue of rats by enzyme-linked immunosorbent assay. Data were processed with one-way analysis of variance and t test. Results: At PIH 72, lung tissue structure of rats in control group was clear and complete, with no inflammatory cell infiltration. At PIH 6, there was edema, hemorrhage, and inflammatory cell infiltration in lung tissue of rats in injury group. At PIH 24, edema, hemorrhage, and inflammatory cell infiltration in lung tissue of rats in injury group aggravated. At PIH 72, area of edema in lung tissue of rats in injury group was enlarged, with obvious hemorrhage and inflammatory cell infiltration. At PIH 6, 24, and 72, pathological score of lung tissue of rats in injury group was (3.43±0.86), (5.39±0.93), and (9.99±0.84) points, respectively, obviously higher than that of rats in control group at PIH 72 [(2.11±0.20) points, t=3.659, 8.450, 22.355, P<0.05]. As time post injury prolonged, pathological scores of lung tissue of rats in injury group were significantly increased (F=121.244, P<0.01). At PIH 6, 24, and 72, expression of NF-κB p65 mRNA in lung tissue of rats in injury group was 15.5±4.3, 25.9±1.8, 30.9±3.5 respectively, significantly higher than that of rats in control group at PIH 72 (7.8±0.8, t=4.315, 20.445, 14.408, P<0.01). As time post injury prolonged, expression of NF-κB p65 mRNA in lung tissue of rats in injury group gradually increased (F=32.691, P<0.01). At PIH 6, 24, and 72, content of TNF-α, IL-1β, and IL-6 in lung tissue of rats in injury group was significantly higher than that of rats in control group at PIH 72, respectively (t=7.650, 8.968, 6.827, 6.726, 8.978, 3.460, 5.420, 13.289, 16.438, P<0.01). At PIH 24, content of TNF-α and IL-1β in lung tissue of rats in injury group was higher than that of rats in the same group at PIH 6 and 72, respectively (t=3.409, -2.549, 4.047, -4.100, P<0.05). At PIH 24 and 72, content of IL-6 in lung tissue of rats in injury group was respectively higher than that of rats in the same group at PIH 6 (t=8.273, 9.711, P<0.05). Conclusions After inhaling white smoke from burning smoke pot, rats are inflicted with lung injury by increasing expression of NF-κB p65 mRNA and content of TNF-α, IL-1β, and IL-6, and induce pathological changes of edema, hemorrhage, and inflammatory cell infiltration of lung tissue.