Objective To study the effects of acetylcysteine magnesium on the vasoactive substances and hepatic fibrosis indexes in liver cirrhosis and portal hypertension of rats.Methods The rat liver cirrhosis model was made with 12 μg/kg dimethylnitrosamines.Then acetylcysteine magnesium was injected respectively with 25,50,and 100 mg· kg-1 dose daily into abdominal cavity.After 8 weeks treatment,pathological section,TGF-β1,NO,TNOS and iNOS of hepatic tissue were detected to assess the effect of acetylcysteine magnesium against cirrhosis portal hypertension.Results After the DMNA modeling was completed,the HE and Sweet reticulocyte staining of liver pathological section showed that cirrhosis of the liver was in the Ⅲ-Ⅳ phase,the infiltration of lymphocytes and formation of pseudolobuli in liver were alleviated in three acetylcysteine magnesium treatment groups (low,medium,and high dose),and the degree of liver fiber sclerosis in three groups was significantly lower than control group.Compared with control group,TGF-β1,NO,TNOS and iNOS were significantly reduced in all treatment groups(P ＜0.05).Conclusion Acetylcysteine magnesium is probably a distinctive antioxidant which can remove various free radical in body and modulate ligand-dependent signal transduction and the growth of cell.It also have protection in the liver cirrhosis and portal hypertension of rats induced by dimethylnitrosamine.

Objective Clinical data of 19 Chinese patients with 21 hydroxylase deficiency (21OHD) were analyzed to improve the diagnosis and treatment level. Methods Clinical features and laboratory data were collected from 19 patients with 21OHD before and after treatment. Results In male patients, the average age of early appearance of secondary sexual character was (9.3?2.8)yrs, and excess androgen resulted in phallic enlargement. Primary amenorrhea was the most common complaint in female(87.5%), and the signs included a varying degree of labioscrotal fusion and clitoral enlargement. The average level of 17-hydroxy progesterone(17OHP) was (63.42?35.07) ?g/L, and adrenocorticotrophic hormone(ACTH), dehydroepiandrosterone(sodium) sulfate(DHEAS) and testosterone(T) were obviously elevated. CT scan showed bilateral adrenal hyperplasia. The level of 17OHP was significantly decreased after treatment[(63.42?35.07) ?g/L vs (3.15?2.71) ?g/L](P

Fruit ripening is associated with a number of physiological and biochemical changes. They include degradation of chlorophyll, synthesis of flavor compounds, carotenoid biosynthesis, conversion of starch to sugars, cell wall solublisation and fruit softening. These changes are brought about by the expression of specific genes. People are interested in the molecular mechanism involved in the regulation of gene transcription during fruit ripening. Many fruit-specific promoters such as PG, E4, E8, and 2A11 have been characterized and shown to direct ripening-specific expression of reporter genes. AGPase plays the key role in catalyzing the biosynthesis of starch in plants. It is a heterotetrameric enzyme with two small subunits and two large subunits, which are encoded by different genes. In higher plants, small subunits are highly conserved among plant species and expressed in all tissues. And the large subunits are present at multiple isoforms and expressed in a tissue-specific pattern. In fruits, the expression pattern of the large subunits varies with plant species. That made it important to study the transcriptional regulation of the large subunits of AGPase in different plant species. Northern-blot analysis indicates in watermelon, an isoform of the large subunits Wml1 expressed specifically in fruits, not in leaves. The 5' flanking region of Wml1, which covers 1573bp, has been isolated through the method of uneven PCR. And transient expression assay has shown that the 1573bp (named WSP) can direct fruit-specific expression of GUS gene. Our goal in this study was to scan the promoter region for main regulatory regions involved in fruit-specific expression. A chimaeric gene was constructed containing the WSP promoter, the beta-glucuronidase (GUS) structural sequence as a reporter gene and the nopaline synthase polyadenylation site (NOS-ter). The plasmid pSPA was digested with Hind III + Hinc II and promoter fragment of 1573bp (from 180bp to 1752bp) was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-16. The same insert was then cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by HindIII + Bgl II to produce pISPA-16. Three 5'-end deletions of the promoter were obtained and fused to GUS gene in plant transient expression vector pBI426: the 1201bp fragment (from 551bp to 1752bp) was generated by digestion of pBSPA-16 with BamH I + SnaB I, the 898bp fragment (from 854bp to 1752bp) by BamH I + EcoRV. Both fragments were ligated with pBluescript SK(-) digested by BamH I + Sma I, to produce pBSPA-12 and pBS-PA-9. The inserts were cut out with HindmIII + BamH I and ligated with pBI426 digested by Hind III + Bgl II, to produce pISPA-12 and pISPA-9. The 795bp fragment (from 957bp to 1752bp) was generated by digestion of pSPA with Hinc II + EcoR I, promoter fragment was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-8. The same insert were cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by Hind III + Bgl II. The 1573bp fragment and three 5'-end deletions were delivered into watermelon leaf, stem, flower and fruit of different development stages (5, 10, 20 days after pollination) via particle bombardment using a biolistic PDS-1000/He particle gun. Bombardment parameters were as follows: a helium pressure of 1200 psi, vacuum of 91432.23Pa, 7 cm between the stopping screen and the plate. Histochemical assay were done on all the tissues bombarded after incubation for 2 days. The 1573bp fragment had the strongest promoter activity, and can induce GUS expression in fruits of 5 and 20 days after anthesis and flowers, but not in fruits of 10 days after anthesis, leaves and stems. Fragments of 1201bp and 898bp can induce GUS expression only in fruits of 20 days after anthesis, and with lower expression levels than 1573bp. Fragment of 795bp was not able to direct GUS expression in any of the tissues bombarded (data not shown). It can be concluded that of the 1573bp, 1201 bp, 898bp Wml1 5'flanking regions include the necessary information directing fruit-specific expression. Deletion from 180bp to 551bp doesn't affect the fruit-specificity of the promoter, but lowered the expression level. There may be some cis-acting elements located in this region, which can enhance external gene expression in later stages of fruit development. Deletion from 854bp and 958bp led to loss of GUS expression. This region includes the necessary information needed for gene expression as well as the regulatory elements for fruit-specific transcription.
Citrullus ; genetics ; Fruit ; genetics ; Gene Expression Regulation, Plant ; genetics ; Promoter Regions, Genetic ; genetics ; Regulatory Sequences, Nucleic Acid ; genetics ; physiology

Objective To establish a renal cell co-culture system to simulate the renal barrier system,and to test its responsiveness to different glucose concentrations,and to investigate the regulatory effect of miR-29b-3p on osteopontin(OPN)/transforming growth factor β(TGF-β)pathway and the changes of this pathway under high glucose condition.Methods The three-cell co-culture system consisting of human renal podocytes,human glomerular mesangial cells and human renal tubular epithelial cells was established to test the cell viability and glucose consumption value at glucose concentrations of 5,8,12 and 16 mmol/L.The content of TGF-β and OPN in cell supernatant was measured.The recombinant plasmid and siRNA of OPN were transfected,and the expressions of TGF-β and OPN were detected by Q-PCR and Western blot.Results The mRNA expressions of OPN,TGF-β and miR-29b were significantly increased at 12 mmol/L glucose conditions.Western blot results showed that the protein expression of OPN increased in high glucose conditions,while the protein expression of TGF-β did not change significantly.After adding miR-29b-3p activator,the mRNA levels of OPN and TGF-β in the cell supernatant were significantly increased.After adding miR-29b-3p inhibitor,the mRNA levels of OPN and TGF-β in the cell supernatant were significantly decreased.Western blot results showed that compared with 5 mmol/L glucose,the protein expressions of OPN and TGF-β were increased by miR-29b-3p activator,and the protein expressions of OPN and TGF-β were decreased by miR-29b-3p inhibitor.After transfection with OPN recombinant plasmid,the content of TGF-β in the cell supernatant was significantly increased,and the mRNA expressions of OPN and TGF-β in the cells were significantly increased.After transfection with OPN siRNA,the content of TGF-β in the cell supernatant was decreased,and the expression of OPN mRNA in the cells was significantly decreased,but the expression of TGF-β mRNA was not significantly increased.Conclusion The renal cell co-culture system can mimic the complex renal environment in vivo.When induced by high glucose,cell proliferation is inhibited,glucose consumption is increased,and the content of TGF-β in the cell supernatant is increased,and miR-29b-3p has a regulatory effect on OPN/TGF-β signaling pathway in the co-culture system.

To construct a recombinant replication-defective human adenovirus type 5 expressing Cap protein of PCV2 and test the immunological efficacy in mice. In this study, the recombinant replication-defective human adenovirus type 5, named as rAd5-Cap (wt-rAd5), was constructed through homologous recombination internally in the HEK293AD cells after co-transfection of the Pac I-linearized backbone plasmid and the shuttle plasmid pacAd5CMV-Cap containing the open reading frame (ORF2) of the porcine circovirus type 2 (PCV2) cap protein or pacAd5CMV without inserted fragment. Furthermore, the rAd5-Cap could induce the expression of PCV2 cap protein in the HEK293AD cells with high efficacy evaluated by the RT-PCR and indirect immunofluorescence assay (IFA). The virus titer of rAd5-Cap could reach up to 10(8.5) TCID50/mL similarly to that of wt-rAd5, indicating that there was little affect on the virus proliferation after the insertion of PCV2 cap protein gene. The humeral immune responses could be activated and detected 14 days after the inoculation of the mice with 10(7) TCID50 rAd5-Cap intramuscularly, and constantly in crease in another 14 days. These molecular biological and animal experiments results demonstrated that the PCV2 cap protein could be efficiently expressed by the recombinant adenovirus rAd5-Cap in eukaryotic cells and induce robust immune responses in mice, which laid a good foundation for the development of new type vaccine against porcine circovirus.
Adenoviruses, Human ; genetics ; Animals ; Antibodies, Viral ; blood ; Capsid Proteins ; genetics ; immunology ; Circovirus ; immunology ; Defective Viruses ; genetics ; HEK293 Cells ; Humans ; Mice ; Recombinant Proteins ; biosynthesis ; immunology ; Virus Replication

OBJECTIVETo analyze the incidence and mortality of oral cavity and pharyngeal cancer in cancer-registration areas of China in 2009.

METHODSWe collected data about incidence of oral cavity and pharyngeal from 72 cancer registry sites of National Central Registry Database in 2009, covering 85 470 522 person (57 489 009 were from urban areas, 27 981 513 were from rural areas).Incidence and mortality rates, proportions, cumulative rate (0-74 years old), cut rate (35-64 years old), age-specific rate were then calculated and analyzed respectively. The age-standardized rate was calculated and adjusted by the Chinese standard population in 1982 as well as the Segi's world standard population.

RESULTSThere were 2803 new diagnosed oral cavity and pharyngeal cancer cases, 1793 male and 1010 female, with the sex ratio at 1.78: 1. The crude incidence rate of oral cavity and pharyngeal cancer was 3.28/100 000(2803/85 470 522). The crude incidence rate of males was 4.15/100 000(1793/43 231 554) while it was 2.39/100 000(1010/42 238 968) among females. The age-standardized incidence rates by Chinese standard population (ASIRC) and the world standard population were 1.72/100 000 and 2.23/100 000 respectively, and the cumulative rate and cut rate was separately 0.26% and 4.02/100 000. The crude incidence and ASIRC of oral cavity and pharyngeal cancers were 3.87/100 000 (2225/57 489 009) and 1.97/100 000 in urban areas, whereas in rural areas, they were 2.07/100 000(578/27 981 513) and 1.17/100 000. There were 1172 death cases, including 825 males and 347 females. The crude mortality rate was 1.37/100 000 (1172/85 470 522), while it was 1.91/100 000(825/43 231 554) among males and 0.82/100 000(347/42 238 968) among females. The age-standardized incidence rates were 0.64/100 000 and 0.88/100 000 respectively, by Chinese standard population (ASMRC) and the world standard population. The cumulative mortality rate (0-74 age years old) and cut rate were separately 0.10% and 1.34/100 000. The mortality and ASMRC were 1.59/100 000(915/57 489 009) and 0.72/100 000 in urban areas, whereas in rural areas, they were 0.92/100 000(257/27 981 513) and 0.48/100 000 respectively.

CONCLUSIONSBoth the incidence and mortality of oral cavity and pharyngeal cancer in China were still low in 2009.

Adolescent ; Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Incidence ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Mouth Neoplasms ; epidemiology ; mortality ; Pharyngeal Neoplasms ; epidemiology ; mortality ; Rural Population ; Sex Distribution ; Survival Rate ; Urban Population ; Young Adult

BACKGROUNDEstrogen deficiency contributes to postmenopausal osteoporosis. Periosteum might be a potential target of estrogen, but the underlying mechanism at gene level is far from being elucidated. The objective of this study was to investigate the correlation between estrogen and fatty acid synthase (FAS) expression in periosteum.

METHODSHuman periosteum cells were cultured in vitro. Expressed genes in the substrated cDNA library were verified using semi-quantitative PCR and real-time PCR. The expression of FAS in periosteum of ovarectomized (OVX) SD rats was investigated.

RESULTSFAS gene was most significantly expressed in the subtracted cDNA library of periosteal cells screened by semi-quantitative PCR. Low FAS expression was verified by real-time PCR in the estrogen exposed human periosteum rather than in the control. The estradiol levels were (20.81 +/- 12.62) pg/ml, (19.64 +/- 4.35) pg/ml and (13.47 +/- 1.84) pg/ml in the sham group, the control, and the OVX group, respectively. The estradiol levels in the OVX group was significantly lower (P = 0.0386). The FAS gene expression in periosteum in the OVX group, sham group, and control group was 3.09 +/- 1.97, 1.33 +/- 0.47 and 1.51 +/- 1.32, respectively. The gene expression in the OVX group was significantly higher (P = 0.0372).

CONCLUSIONEstrogen modulates FAS gene expression in in vitro human perisoteum as well as in in vivo rat periosteum.

Animals ; Cells, Cultured ; Estradiol ; blood ; pharmacology ; physiology ; Fatty Acid Synthases ; genetics ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Ovariectomy ; Periosteum ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the association of spermatogenic arrest with the expression of estrogen receptor alpha (ERalpha) in human testes.

METHODSWe examined the testicular biopsy specimens of 120 infertile men by HE staining, detected the expression of ERalpha in the specimens of those with spermatogenic arrest by the two-step immunohistochemical method, and compared the results with those of 10 healthy men.

RESULTSOf the 120 specimens from the infertile men, 31 (25.8%) met the diagnostic criteria of spermatogenic arrest. In the testis tissue of normal men, ERalpha expressed in Sertoli, myoid and Leydig cells, but not in spermatogenic cells, while in the testis tissues of those with spermatogenic arrest, ERalpha expressed lowly in Sertoli, myoid and Leydig cells, with statistically significant differences in immunostaining intensity between the two groups (P < 0.05).

CONCLUSIONAndrogen receptor (AR) and ERalpha may play a coordinating role in facilitating spermatogenesis. Spermatogenic arrest may be related to a complex series of disorders in cell signal transduction involving AR, ERalpha and HSP90.

Adult ; Case-Control Studies ; Estrogen Receptor alpha ; metabolism ; Humans ; Infertility, Male ; etiology ; metabolism ; pathology ; Male ; Spermatogenesis ; Testis ; metabolism ; pathology ; Young Adult

Objective To discuss the optimal clinical diagnosis and treatment of ectopic ACTH syndrome with occult tumors. Methods Clinical features, imaging examinations and treatment of 17 patients with ectopic ACTH syndrome were described and compared. Results All patients illustrated the typical clinical features of Cushing’s syndrome. They had hypokalemic alkalosis, elevated serum cortisol and plasma ACTH levels. In the high-dose dexamethasone suppression tests, most patients failed to suppress serum cortisol and 24-hour urinary cortisol. CT and MRI are useful imaging modalities to localize the ACTH-secreting tumor in patients with ectopic ACTH syndrome. The patients with overt ACTH-secreting tumors had surgical curative resection soon after diagnosis. Among patients with occult ACTH-secreting tumors, three underwent subtotal bilateral adrenalectomy, two underwent right adrenalectomy, four received inhibitor of steroidogenesis aminoglutethimide. Their hypercortisolism was controlled. Conclusion Surgical curative resection is the optimal treatment of ectopic ACTH syndrome with overt ACTH-secreting tumor. Bilateral adrenalectomy, right adrenal ectomy or chemotherapy to control hypercortisolism is an available treatment of ectopic ACTH syndrome with occult ACTH-secreting tumors.

Objective To evaluate the predictive value of baseline serum high sensitivity C-reactive protein for the first cardio-cerebral vascular event in the population with diabetes. Method In this prospective cohort study, a total of 101 510 employees of Kai Luan Group, who received healthy examination from July 2006 to October 2007, were screened and 7865 subjects with fasting plasma glucose ≥ 7. 0 mmol/L or known diabetes mellitus and under insulin or hypoglycemic drugs therapy were followed up for 38 - 53 (48. 02 ± 3. 14) months. Results ( 1 ) Incidence rates of total cardio-cerebral vascular events, cerebral infarction and myocardial infarction increased in proportion to increased levels of baseline hsCRP ( P ＜ 0. 01 ). After adjusting for age, gender, body mass index ( BMI), systolic blood pressure( SBP), diastolic blood pressure (DBP), total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and cigarette smoking, multivariate Cox's proportional hazards regression analysis indicated that the individuals in the highest quartile of hsCRP levels group (hsCRP≥2. 50 mg/L) had an increased risk of total cardio-cerebral vascular events (RR: 1.64, 95% CI:1.20-2.24), cerebral infarction (RR: 1.52, 95% CI: 1.03-2.24), myocardial infarction (RR: 2.57,95% CI: 1.34 -4. 91 ) compared with those in the lowest quartile group( hsCRP ＜ 0. 41 mg/L). (2) Higher baseline hsCRP levels were associated with aging, female gender, higher BMI, SBP, DBP, fasting blood glucose, TC, TG, LDL-C levels and lower HDL-C levels ( all P ＜ 0. 05 ). Conclusion Baseline hsCRP level is associated with increased first cardio-cerebral vascular event in the population with diabetes.