AIM: To study the molecular mechanism of reproduction dysfunction in pubertal diabetic rat, the level of 5?-reductase (type 2) mRNA in testis, epididymis and prostate in diabetic rat was detected. METHODS: The gene expression of 5?-reductase (type 2) was detected by Northern blot. RESULTS: In the caput of the epididymis, the expression of 5?-reductase (type 2) mRNA of D and ID groups was less than that of C group. CONCLUSION: The decreased expression of 5?-reductase (type 2) results in the decreased production of dihydrotestosterone, which influences the development and function of reproduction system in pubertal rats. [

Objective To establish a capillary gas chromatography method for determination of camphol and isoborneol in Shaoshang yuhe gao ( burn healing cream) . Methods The capillary gas chromatography was adopted under the following conditions: use PEG-2000 as the stationary liquid,nitrogen as carrier gas,ZB-WAX (30 m×0. 25 mm,0. 25 μm) as the chromatographic column,and the flame ionization detector. The column temperature was programmed at 80 ℃ for 5 min as the initial temperature,then raised to 180 ℃ at the rate of 5℃·min-1 and kept for 10 min. The shunt ratio was 101. Results The liner range for camphol was 0. 487 5-31. 25 μg ( r =0. 999 6),and the average recovery was 95. 95%( n =6). The liner range for isoborneol was 0. 487 5-31. 25 μg( r =0. 999 7),and the average recovery was 96. 44%( n =6). Conclusion The method is accurate,sensitive,and can be applied to quality control of shaoshang yuhe gao.

Objective To investigate the inhibitory effects of RNA interference on expression of matrix metalloproteinase-2(MMP-2)gene and invasiveness of human pancreatic cancer cell line PANC-1 in vitro.Methods Small interference RNA targeting MMP-2 gene was designed and constructed to plasmid pGCsi-U6.Recombinant plasmids were transfected to pancreatic carcinoma PANC1 cells with Lipofectamine 2000.The efficiency of transfection was evaluated by flow cytometry.RQPCR was used to detect the expression of MMP-2 mRNA.The expression of MMP-2 protein was determined by ELISA.The invasiveness of PANC-1 cells was measured by transwell chamber experiment.MTT assay was used to detect the proliferation and growth of PANC-1 cells.Results Sequencing confirmed that the MMP-2 siRNA plasmid was successfully constructed.The best efficiency of transfecting recombinant plasmid was 82.1%.After transfection of the MMP-2 siRNA plasmid, the MMP-2 gene expression of PANC-1 cells was suppressed to 71.74 %(P＜0.05),and protein expression of MMP-2 fell to 49.82%(P＜0.05).The corresponding inhibition ratio of invasiveness was 33.0%(P＜0.05).There was no marked difference in proliferation rate measured by MTT assay among different groups(P＞0.05).Conclusions RNAi targeting MMP-2 can suppress invasiveness of PANC-1 cells in vitro.This suggests that MMP-2 could be a target for gene therapy of pancreatic carcinoma.RNAi is expected to open up a new prospect for tumor therapy.

Objective To observe the expression of neuron specific enolase (NEC) to evaluate the neuroprotective effect of a cell-penetrating Bax-inhibiting peptide (BIP) on neonatal rats with hypoxic-ischemic brain damage (HIBD). Methods Wi-star rats (7-day old) were randomly divided into Sham group, BIP group and HIBD group. After modeling HIBD, the histologi-cal (HE staining) and immunohistochemistry methods were used to determine the apoptotic pathological changes and the NSE expression levels in the brain at different time points. Results Compared to the Sham group, the rats of HIBD group showed significant apoptotic pathological changes. The histological changes and the brain damages were improved significantly in BIP group at each sampling point. The number of NSE-positive cells was significantly decreased in HIBD and BIP groups over time (P<0.05). The number of NSE-positive cells had significant difference among different groups at 48 h, 96 h and 7 d after opera-tion (F=45.35-81.66, P<0.01). The number of NSE-positive cells in the HIBD group was smaller than that of the Sham group and BIP group 48 h after operation (P<0.05). The number of NSE-positive cells in the BIP group was smaller than that of the Sham group 96 h after operation (P<0. 05). Conclusions BIP can decrease the apoptosis of cortex nerve cells in 7-day old HIBD rat model, and may have neuroprotective effect on the early stage of HIBD.

In recent years, with the development of sequencing technology and research on molecular changes in different races of prostate cancer, it has been found that the pathogenesis of prostate cancer showed obvious ethnic differences. This article reviewed the research progress of ETS fusion genes, FOXA1, SPOP, IDH1 and other driver genes in prostate cancer. And the research of molecular typing of prostate cancer showed different patterns of molecular changes in Chinese and western populations. The molecular changes of prostate cancer in western populations were dominated by ETS fusion gene, while those in Chinese populations were dominated by gene mutations, mainly FOXA1 and SPOP mutations. Moreover, the dominant fusion gene in Chinese prostate cancer was not ETS fusion gene, but SCHLAP1-UBE2E3.

OBJECTIVE: To investigate the effects of Fuzheng Huayu Recipe on renal interstitial fibrosis in rats induced by the toxic substances. METHODS: Forty Wistar rats were divided into 3 groups, including the normal group, model control and Fuzheng Huayu Recipe treated group. The renal fibrotic model was induced with intraperitoneal injection of dimethylnitrosamine (DMN) for 4 weeks and oral administration of food containing heavy metals such as Hg, etc for 8 weeks. From the 5th week, the rats were orally administered Fuzheng Huayu Recipe for 4 weeks. The renal tissue was stained with HE and periodic acid-silver metheramine, respectively. The serum creatinine and urea nitrogen were assayed with the kits, and the renal hydroxyproline (Hyp) contents in the homogenate of kidney tissue were measured with HCl hydrolysis. RESULTS: In the model group, the renal tubule was swollen, and the tubular epithelial cells were degenerated, necrosed and detached, with the infiltration of inflammatory cells; the tubular interstitium was expanded, characterized with the deposition of large collagens; the tubular basement membrane became thicker and wrinkled; and the renal Hyp content and serum creatinine and urea nitrogen levels increased remarkably as compared to the normal group. After the treatment with Fuzheng Huayu Recipe, the renal tubular inflammation and interstitial collagen deposition were alleviated, and the serum creatinine and urea nitrogen levels decreased significantly as compared to the model group. CONCLUSION: Administration of DMN and heavy metals can induce the renal interstitial fibrosis and impair the renal function. Fuzheng Huayu Recipe can improve the impaired renal functions and reverse the renal interstitial fibrosis.

OBJECTIVETo demonstrate the side effects of malariotherapy and to explore safe procedures in conduct of malariotherapy for human immunodeficiency virus (HIV) infected patients.

METHODSTwenty HIV/acquired immunodeficiency syndrome (AIDS) patients were selected for the study of malariotherapy and were intravenously infected with Plasmodia vivax to induce therapeutic malaria. Malaria was terminated with chloroquine after 10 - 20 malarial febrile episodes. Clinical assessments were made before (baseline), during (malarial phase) and after (post) termination of malaria. The density of Plasmodia in peripheral blood from the HIV/AIDS patients were compared to that from HIV-negative naturally infected malarial patients who donated the blood for the therapeutically induced malaria. CD(4) cell baseline levels were correlated to the severity of malarial symptoms and parasitemia.

RESULTSThere were no significant differences of Plasmodium density between the HIV/AIDS patients injected with P. vivax and the HIV-negative blood donors. However, it was found that the HIV-positive patients had milder malarial symptoms and parasitemia with progressively lower CD(4) cell baseline levels. All patients developed every day or every other day fever episodes with headache and shaking chill. These symptoms were well tolerated with the aid of anti-pyretic medications. Spleen and liver enlargement were seen in 15 of 20 and 4 of 20 patients respectively. Transitory liver effects with increase of serum glutamic-pyruvic transaminase were seen in 2 of 20 during malarial phase. Most patients experienced mild to medium anemia and 6 of 20 patients developed thrombocytopenia during malarial phase. All these side effects disappeared after termination of malaria or within one month thereafter. No complications occurred in these patients.

CONCLUSIONSTherapeutically induced acute vivax malaria was well tolerated in 20 HIV-positive subjects who represented a range of CD(4) cell levels from 1868/ micro l to 15/ micro l. Malariotherapy did not induce complications while increasing CD(4) cell levels in most treated HIV/AIDS patients (results published elsewhere).

Adult ; Animals ; CD4-Positive T-Lymphocytes ; Female ; HIV Infections ; therapy ; Humans ; Immunotherapy ; adverse effects ; methods ; Lymphocyte Count ; Malaria, Vivax ; immunology ; Male ; Plasmodium vivax ; immunology

Objective: To investigate the effects of lactoprotein iron chelates on rats with iron deficiency anaemia (IDA), so as to provide insights into developing and utilizing novel iron supplements. Methods: Seventy weaning female SPF-graded rats of the SD strain were randomly divided into the control group (A), model group (B), ferrous sulfate group (C), lactoferrin group (D), lactoferrin iron chelate group (E), Casein oligopeptide iron chelate group (F) and whey protein oligopeptide iron chelate group (G), with 10 rats in each group. The rats in group A were fed with normal diet, and the others were fed with poor iron diet for IDA modeling. The corresponding interventions were given by intragastric administration once a day. The iron ion concentrations of group C, E, F and G were 2.0 mg/kg, and the protein and oligopeptide concentrations of group D, E, F and G were 2 000 mg/kg. Body weight and hemoglobin of rats were measured weekly during 21-day intervention. At the end, peripheral blood samples were collected, and blood routine, iron metabolism and liver function indicators were determined. Results: After the intervention, among blood routine indicators, the rats in group C, E, F and G showed elevated hemoglobin, red blood cell, mean corpuscular volume and hematocrit, and decreased free protoporphyrin and mean corpuscular hemoglobin concentration when compared with the rats in group B (all P<0.05); among iron metabolism indicators, the rats in group C, E and G showed elevated serum ferritin, the rats in group C, E, F and G showed elevated serum iron, the rats in group C, D, E, F and G showed decreased unsaturated iron binding capacity and total iron binding capacity when compared with the rats in group B (all P<0.05); among liver function indicators, the rats in group E and G showed decreased alanine transaminase when compared with the rats in group B (both P<0.05). Conclusions Lactoprotein alone could not completely improve IDA in rats compared with traditional iron supplement (ferrous sulfate). Lactoprotein iron chelate, especially whey protein oligopeptide iron chelate, could significantly improve IDA, iron reserve and liver function damage in rats.

Background and Objectives: Chronic obstructive pulmonary disease (COPD) is a common, frequently-occurring disease and poses a major health concern. Unfortunately, there is current no effective treatment for COPD, particularly emphysema. Recently, experimental treatment of COPD using mesenchymal stem cells (MSCs) mainly focused on bone marrow-derived MSCs (BM-MSCs). Human umbilical cord-derived MSCs (hUC-MSCs) have more advantages compared to BM-MSCs. However, studies on the role of hUC-MSCs in management of COPD are limited. This study sought to explore the role of hUC-MSCs and its action mechanisms in a rat model of VEGF receptor blocker SU5416-injured emphysema. Methods: and Results: hUC-MSCs were characterized by immunophenotype and differentiation analysis. Rats were div-ided into four groups: Control, Control＋MSC, SU5416 and SU5416＋MSC. Rats in model group were administered with SU5416 for three weeks. At the end of the second week after SU5416 administration, model group were infused with 3×106 hUC-MSCs through tail vein. After 14 days from hUC-MSCs transplantation, rats were euthanized and data were analyzed. HE staining and mean linear intercepts showed that SU5416-treated rats exhibited typical emphysema while emphysematous changes in model rats after hUC-MSCs transplantation disappeared completely and were restored to normal phenotype. Furthermore, hUC-MSCs inhibited apoptosis as shown by TUNEL and Western blotting.ELISA and Western blotting showed hUC-MSCs rescued VEGF-VEGFR2-AKT pathway in emphysematous lungs. Conclusions The findings show that hUC-MSCs effectively repair the emphysema injury. This study provides the first evidence that hUC-MSCs inhibit apoptosis via rescuing VEGF- VEGFR2-AKT pathway in a rat model of emphysema.

OBJECTIVETo explore the mechanisms of malariotherapy for human immunodeficiency virus (HIV)-infected patients and to identify which stage(s) of HIV infection is suitable for the treatment of malariotherapy.

METHODSTherapeutic acute vivax malaria was induced and terminated after 10 fever episodes in 12 HIV-1-infected subjects: Group 1 (G1) had 5 patients with CD(4) T-cell counts >or=500/ micro l at baseline, Group 2 (G2) had 5 patients with CD4 at 499 - 200/ micro l and Group 3 had 2 patients with CD(4) < 200/ micro l (not included in statistical analysis). Enzyme-Linked-Immuno-Sorbent Assay (ELISA) was used to measure plasma levels of cytokines and soluble activation markers. Flow cytometry was used to measure levels of lymphocyte subsets and phenotypes and CD(4) cell apoptosis. Bayer bDNA assay was used to test plasma levels of HIV-1 RNA (viral load). Samples were taken and tested twice before malaria (baselines), three times during malaria and seven times after termination of malaria (at day 10 and 1, 3, 6, 12, 18 and 24 months).

RESULTSLevels of plasma tumor necrosis factor-alpha (TNF-alpha), soluble TNF-alpha receptor-2 (sTNF-RII), neopterin (NPT) and soluble IL-2 receptor (sIL-2R) significantly increased during malaria and sharply reduced to baselines post malaria in all groups. Stronger responses of the aforementioned factors were seen in G2 than in G1 during malaria (P = 0.081, 0.001, 0.013, 0.020). CD4 count and percentage; CD(4)/CD(8) ratio and CD(25)(+) and CD(4)(+)CD(25)(+) percentages increased but HLA-DR+ percentage decreased either during or post malaria in G2. Most G2 patients experienced sustained increase but most G1 patients underwent natural history decline of CD(4) counts and percentages during 2-year follow-up. Percentage of apoptotic CD(4) cells decreased post malaria in all groups. G3 patients had weaker immune responses, however, one advanced AIDS patient in this group experienced clinical improvement after malariotherapy. Most of the 12 patients experienced increase of HIV viral load during malaria but the viral load returned to baseline levels 1 - 3 months after cure of malaria and remained near baseline levels for up to two years.

CONCLUSIONSPart of the mechanisms of malariotherapy is to induce high levels of cytokine activities and subsequently the changes of T-lymphocyte subsets and phenotypes in HIV-infected patients. These findings suggest that malariotherapy may treat HIV-1-infected patients whose CD4 baselines are in the range of 500 - 200/ micro l.

Acute Disease ; Adult ; CD4 Lymphocyte Count ; Cytokines ; blood ; Female ; HIV Seropositivity ; immunology ; therapy ; virology ; HIV-1 ; isolation & purification ; Humans ; Malaria, Vivax ; immunology ; Male ; Viral Load