OBJECTIVETo screen the proteins interacting with inhibitor of differentiation 1(Id1) using yeast two-hybrid analysis in adult human lung cDNA libraries.

METHODSThe coding sequence of Id1 was amplified by PCR and cloned into the bait plasmid. The recombinant bait vector pHybLex/Zeo-Id1 was verified by restriction endonuclease digestion before transformation into the yeast strain EGY48/pSH18-34, which was tested subsequently for reporter genes Leu2 and LacZ activation. The pHybLex/Zeo-Id1 plasmid and the cDNA library plasmid were sequentially transformed into the yeast strains and screened to obtain Leu2(+) and Leu2(+)LacZ(+) clones, with the false positive clones excluded using positive and negative controls, and the plasmid of the true positive clone was sequenced and blasted for homological analysis.

RESULTSSuccessful construction of pHybLex/Zeo-Id1 was confirmed by enzyme digestion. After transformation of pHybLex/Zeo-Id1 into EGY48/pSH18-34, no specific reporter genes Leu2 and LacZ activation was found. The pHybLex/Zeo-Id1 plasmid and the cDNA library plasmid were sequentially transformed into yeast strain, and 198 Leu(+) clones and 19 Leu(+)LacZ(+) double positive clones were obtained. After elimination of the false positive clones, one true positive clone was obtained, whose plasmid analysis by sequencing and blasting indicated high homology (99.5%, 556/559) to AGGF1 (an angiogenic factor with G-patch and FHA domains 1). AGGF1 expression was confirmed in the true positive yeast cells by Western blotting.

CONCLUSIONAGGF1 is confirmed to interact with Id1 by yeast two-hybrid analysis for screening adult human lung cDNA libraries.

Adult ; Angiogenic Proteins ; genetics ; metabolism ; Cell Proliferation ; Endothelial Cells ; cytology ; metabolism ; Gene Library ; Humans ; Inhibitor of Differentiation Protein 1 ; metabolism ; Lung ; cytology ; Plasmids ; genetics ; Protein Binding ; Two-Hybrid System Techniques

Objective To investigate the expression of Mta-1 in bladder transitional cell carcinoma (BTCC)and to analyze its correlation with the clinical staging,pathologic grading,metastasis and recur- rence,and to explore the possible molecular mechanisms.Methods Samples of 42 cases of BTCC and 12 normal bladder mueosa tissues were examined with immunohistochemical analysis for the expression of Mta- 1,ER,u-PA and PAI-1.Endothelial cells were stained by anti-CD34,and microvascular density(MVD)of carcinoma tissue was calculated.The correlation of Mta-1 expression with the invasion,metastasis,angiogene- sis and recurrence of BTCCs was analyzed;and the correlation of Mta-1 expression with ER,u-PA,PAI-1,and MVD was also analyzed.Results The positive rate of Mta-1 expression in BTCCs was 73.8%(31/42) , while it was 0.0% in normal bladder mucosa tissues(P＜0.01).The expression level of Mta-l increased with the higher clinical stages and pathologic grades of BTCCs;it was higher in recurrence group(100.0% , 15/15)than in non-recurrence group(59.3%,16/27),and high in metastasis group(100.0%,14/14) than in non-metastasis group(60.7%,17/28)(P＜0.05).The expression level of ER increased with the lower clinical stages and pathologic grades of BTCCs;the positive rate of ER expression was 0.0% in 14 ca- ses with metastasis and was 53.6% in 13 of 28 cases without metastasis(P＜0.05);and the rate was 6.7% in 1 of 15 cases with recurrence and 44.4% in 12 of 27 cases without recurrence(P＜0.05).Negative cor- relation was found between Mta-1 and ER expression(r=-0.739,P＜0.01).The positive rate of u-PA ex- pression(59.5%,25/42)was significantly higher in BTCCs than that in normal bladder mucosa tissues (16.7%,2/12)(P＜0.05).Positive correlation was found between u-PA and Mta-1 expression(r= 0.875),while negative correlation was found between u-PA and PAI-1 expression(r=-0.535).The posi- tive rate of PAI-1 expression in normal bladder mucosa tissues(50.0%,6/12)was significantly higher than that in BTCCs(19.0%,8/42)(P＜0.05).In addition,negative correlation was found between PAI-1 and Mta-1 expression(r=-0.706).And positive correlation was found between MVD in BTCCs marked by an- ti-CD34 and Mta-1 expression(r=0.683).Conclusions Mta-1 is highly expressed in BTCCs,and it correlates closely with tumor pathologic grades,clinical stages,recurrence and metastasis.Mta-1 up-regulates the expression of u-PA and down-regulates that of PAI-1,which is associated with invasion and metastasis and acts as an angiogenic mediator in BTCCs.A negative correlation is found between Mta-1 and ER in inva- sion and metastasis of BTCCs.

OBJECTIVETo study the expression of nNOS and ultrastructural changes in the penile tissue of rats with prolactinoma-induced erectile dysfunction (ED).

METHODSWe established the model of prolactinoma in 20 male Westar rats by peritoneal injection of diethylstilbestrol (DES) and treated the control rats with normal saline (n = 10) or sterilized arachis oil (n = 10). After 8 weeks, we performed the apomorphine test and measured the weight of the pituitary gland and the levels of serum prolactin (PRL) and testosterone (T) to confirm the successful construction of the prolactinoma-induced ED model. Then we determined the expression of nNOS in the penile tissue by immunohistochemistry and examined the ultrastructural changes of the penile cavernosum under the transmission electron microscope.

RESULTSThe prolactinoma-induced ED model was successfully established in 15 rats. The weight of the pituitary gland was significantly increased in the rats treated with DES as compared with the normal saline and sterilized arachis oil controls ([46.7 ± 15.5] vs [11.7 ± 2.4] and [12.4 ± 2.3] mg, both P < 0.05). The level of serum PRL was markedly higher while that of T remarkably lower in the former than in the latter two groups ([1,744.9 ± 304.5] vs [11.5 ± 2.4] and [10.6 ± 1.9] ng/ml, both P < 0.0l; [1.54 ± 0.46] vs [3.11 ± 1.08] and [3.04 ± 1.11] ng/ml, both P < 0.05). The rate of penile erection was significantly reduced in the prolactinoma-induced ED model rats in comparison with the normal saline and arachis oil controls (16.7% vs 100% and 87.5%, both P < 0.05), and so was the expression of nNOS in the penile tissue (0.024 ± 0.011 vs 0.066 ± 0.019 and 0.058 ± 0.021, both P < 0.05). Transmission electron microscopy manifested significant ultrastructural changes in the endothelial and smooth muscle cells of the cavernous tissue in the prolactinoma-induced ED models.

CONCLUSIONThe ultrastructural changes of the penile cavernous tissue and the reduced expression of nNOS in penile tissue may be the most important mechanisms of prolactinoma-induced ED in rats.

Animals ; Apomorphine ; Carcinogens ; Diethylstilbestrol ; Erectile Dysfunction ; etiology ; Humans ; Male ; Myocytes, Smooth Muscle ; ultrastructure ; Nitric Oxide Synthase Type I ; metabolism ; Organ Size ; Penile Erection ; Penis ; enzymology ; ultrastructure ; Pituitary Neoplasms ; chemically induced ; complications ; Prolactin ; blood ; Prolactinoma ; chemically induced ; complications ; Rats ; Rats, Wistar ; Testosterone ; blood

OBJECTIVETo evaluate the vulnerability to floods in Guangdong province at district level.

METHODSData were collected from the sixth census, the 2010 Statistical Yearbook of Guangdong, the 2010 Health Statistics Yearbook of Guangdong and China Disease Prevention and Control information systems, etc. The weight of each indicator was determined based on subjective method and objective method respectively; and finally the results of the two methods were compared.

RESULTS13 indicators were selected for the assessment of vulnerability to floods, including 6 sensitivity indicators, 5 adaptability indicators and 2 exposure indicators. Indicators with large weight (subjective weight/objective weight) were the proportion of population older than 65 years old (0.31/0.30), the proportion of population older than 65 years old (0.16/0.23), infant mortality rate (0.18/0.20), the total Gross Domestic Product (GDP) per capita (0.33/0.21), the proportion of illiterate in the population older than 15 years old (0.19/0.28), history frequency of floods (0.75/0.75). The mean vulnerability index (VI) calculated by subjective method was 0.35 with the standard deviation of 0.10; the mean vulnerability index calculated by objective method was 0.31 with the standard deviation of 0.08. The two weighting methods showed consistent results of vulnerability index (ICC = 0.975, P < 0.01). VI of most districts dropped in the interval of 0.30 - 0.39. Districts with subjective VI > 0.50 or objective VI > 0.40 should pay more attention to floods, including parts of the coastal areas, Beijiang River Basin, the eastern tributary area of Dongjiang River and the northern part of Pearl River Delta. Dapu district of Meizhou (0.55/0.45), Dianbai district and Maogang district of Maoming (0.54/0.48) were most vulnerable. Districts of Heyuan, Dongguan, Zhaoqing and Huizhou were less vulnerable, Yuancheng district of Heyuan showed least vulnerable to floods (0.15/0.12) followed by Dongguan (0.18/0.16), Duanzhou district (0.18/0.16) and Guangning (0.17/0.15) district of Zhaoqing. The score of indicators differed among different level of vulnerability (P < 0.05).

CONCLUSIONDifferent regions of Guangdong province showed different vulnerability to floods, vulnerable areas should be priority in the prevention and control of floods.

China ; Climate ; Demography ; Disasters ; Floods ; Humans ; Risk Assessment ; Rivers

Qumazi is a commonly used Tibetan medicine. With a long history, it can be found in the Four Medical Tantras written by gYu-thog rNying-ma Yon-tan mGon-po since the 8th century AD. Qumazi grows in mudflats and fields, including species growing in highlands, lowlands, mountains and farmlands. According to records in Crystal Beads Materia Medica, it features green sword-shaped leaves, thin stems with red veins, inserted panicles, white chicken-like flowers and copper needle row-like roots. However, there are many inconsistent morphological descriptions for Qumazi plants in many Chinese versions of Tibetan medicine books. In this article, after studying ancient and modern Tibetan medicine books, consulting experts and conducting surveys, the authors confirmed that Qumazi belongs to Rheum of Polygonaceae, including Rheum nobile Hook. f. et. Thoms, R. globulosum Gage, R. alexandrae Hook. f. et. Thoms, R. pumilum Maxim and R. delavayi Franch. In some regions, Qumazi is substituted by R. spiciforme Royle and R. przewalskyi Losinsk. After the Chinese version of Qinghai-Tibet Plateau Drug Illustrations was published in 1972, Qumazi has been miswritten as P. sibiricum Laxm in many Chinese versions of Tibetan medicine books, perhaps because P. sibiricum Laxm has many similar features with Qumazi as described in Crystal Beads Materia Medica and then is mistranslated from Tibetan to Chinese versions. According to records, Qumazi can reduce edema and is mainly applied to treat the minamata disease in clinic.
China ; History, Ancient ; Medicine, Tibetan Traditional ; history ; Plants, Medicinal ; chemistry ; growth & development ; Polygonaceae ; anatomy & histology ; chemistry ; growth & development ; Reference Books, Medical

OBJECTIVE: To compare the clinical features and follow-up results of systemic lupus erythematosus (SLE) between boys and girls. METHODS: A retrospective analysis was performed for the clinical data of 79 children (18 boys and 61 girls), aged ≤14 years, who were diagnosed with SLE from 2008 to 2018. The boys and the girls were compared in terms of initial and major clinical symptoms, injury of organs/systems, related laboratory markers, and follow-up results. RESULTS: As for the initial and non-initial symptoms, fever had the highest incidence rate in the boys, while facial erythema had the highest incidence rate in the girls. The boys tended to develop renal injury and hematological damage (P<0.05), with a significantly higher incidence rate of proteinuria than the girls (P<0.05), while the girls tended to develop joint pain (P<0.05). There were high abnormal rates (>80%) of anti-nuclear antibody, dsDNA, complement C3, and erythrocyte sedimentation rate in both boys and girls (P>0.05). The boys had a significantly higher disease activity than the girls at the first visit and in year 9 of follow-up (P<0.05). A one-month to ten-year follow-up showed that among the boys, 3 were lost to follow-up, 1 died, 7 were well controlled but required oral administration of large doses of hormones or immunosuppression, 2 progressed to chronic renal failure, and 1 developed lupus encephalopathy. Among the girls, 3 were lost to follow-up; 5 died; 34 were well controlled, among whom 5 were maintained on oral prednisone acetate with a dose of <10 mg, 1 was withdrawn from the drug for 1 year, and 2 were withdrawn from the drug for 2 years; 4 developed lupus encephalopathy; 1 developed depression and anxiety and had suicidal tendency in the 7th year after disease onset; 2 experienced impaired vision, blurred vision, and chloropsia; 1 developed a vascular necrosis of both femoral heads in the 3rd year of hormone administration. CONCLUSIONS There are differences in clinical features, several laboratory markers, and prognosis between boys and girls with SLE. Boys tend to have a high severity at disease onset, develop renal injury and hematological damage, and have poor long-term prognosis, while girls tend to have joint involvement.

Objective To construct the orthotopic mouse liver transplantation model and cover troubleshooting,in order to provide experimental techniques support for organ transplantation pathology and immunology studies.Methods Male C57BL/6 mice,10-12 weeks,were selected as the allograft donors.Male C3H mice with same age were selected as the allograft recipients.The orthotopic mouse liver transplantation model consisted of 3 stages,including harvesting the donor liver,back-table preparation of the liver graft and transplantation of the donor liver into the recipient.The average time for harvesting the donor livers was (40 ± 8.8) min,(23 ± 4.7) min for preparing the donor livers and (75 ± 9.6) min for transplanting the donor livers into the recipient.Results Seventy pairs of mice were used for the preliminary experiments.For the formal experiments,the allograft transplantation was established on 220 pairs with 90.4％ successful rate.Conclusion It is the skillful and high quality microsurgical technique that is the guarantee of establishing the orthotopic mouse liver transplantation model successfully.

OBJECTIVETo explore the efficacy of frozen-thawed embryo transfer combined with intrauterine administration of autologous peripheral blood mononuclear cells (PBMCs) in the treatment of repeated implantation failure (RIF).

METHODSPBMCs obtained from 3 patients with RIF on the day of follicle rupture (natural cycle) or when the endometrial thickness reached 8 mm (hormone replacement cycle) were cultured in the presence of HCG for 48 h. The cultured PBMCs, along with freshly isolated PBMCs, were administered into the uterine cavity of the patients. Vitrified cleavage-stage embryos or blastocysts transfer was performed on day 3 or 5, respectively.

RESULTSVitrified embryo or blastocyst transfer resulted in pregnancy and healthy live births in all the 3 patients.

CONCLUSIONFrozen-thawed embryo transfer combined with intrauterine administration of autologous PBMCs may be an effective and safe approach to the treatment of RIF and may improve the outcomes of assisted reproduction.

Adult ; Blastocyst ; Cryopreservation ; Embryo Implantation ; Embryo Transfer ; methods ; Female ; Fertilization in Vitro ; methods ; Humans ; Monocytes ; Pregnancy ; Pregnancy Rate ; Treatment Failure

OBJECTIVETo analyze serum vancomycin concentration after administration of different therapeutic doses in children with Staphylococcus aureus pneumonia (SAP) in order to determine the appropriate dose of vancomycin in clinical administration.

METHODSThe clinical data of 35 children who were diagnosed with SAP and treated with vancomycin from January 2008 to December 2013 were retrospectively analyzed.

RESULTSAmong the 35 SAP cases with vancomycin therapy, 22 cases (63%) had serum vancomycin trough concentration monitored. The numbers of cases with vancomycin at 10, 12.5, and 15 mg/(kg·dose) × every 6 hours (q6h) were 11, 4 and 7, respectively. The mean serum trough concentration of vancomycin in the 15 mg/(kg·dose) group was 14.98 mg/L, which was significantly higher than in the 10 mg/(kg·dose) and 12.5 mg/(kg·dose) groups (4.97 and 8.00 mg/L respectively; P<0.05). The percentage of cases that reached the expected trough concentration in the 15 mg/(kg·dose) group (71%) was significantly higher than that in the 10 mg/(kg·dose) group (9%), but there was no significant difference in this percentage between the 15 mg/(kg·dose) and 12.5 mg/(kg·dose) groups (71% vs 25%).

CONCLUSIONSThe reasonable dosage of vancomycin for the treatment of pediatric SAP is 15 mg/(kg·dose) × q6h or 60 mg/(kg·d).

Adolescent ; Anti-Bacterial Agents ; blood ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Pneumonia, Staphylococcal ; blood ; drug therapy ; Vancomycin ; adverse effects ; blood

OBJECTIVETo compare the clinical manifestations between inhaled and blood-borne Staphylococcus aureus pneumonia (SAP) and the antibiotic resistance between the isolates of inhaled and blood-borne Staphylococcus aureus.

METHODSThe clinical data of 44 pediatric SAP cases in the Children′s Hospital, Chongqing Medical University from January 2008 to December 2013 were retrospectively analyzed. Twenty-four cases were identified as inhaled SAP, and 20 cases as blood-borne SAP.

RESULTSInhaled SAP was more common in children younger than 3 years of age, while blood-borne SAP was more prevalent in children older than 6 years of age. Patients with inhaled SAP had significantly higher incidence rates of cough, wheeze, moist rales, dyspnea and empyema than those with blood-borne SAP (P<0.05). The patients with blood-borne SAP were more vulnerable to severe fever, unconsciousness, dysfunction of liver and kidney, pyogenic osteomyelitis, septic arthritis, sepsis, and abscess of skin and soft tissues (P<0.05). Inhaled SAP isolates had significantly higher rates of resistance to amoxicillin/clavulanic acid, oxacillin, and cefoxitin than blood-borne SAP isolates (P<0.05), while the latter had a higher rate of resistance to cotrimoxazole (P<0.05).

CONCLUSIONSInhaled SAP often occurs in children younger than 3 years of age, and the respiratory manifestations are commonly seen. Blood-borne SAP often occurs in children older than 6 years of age, with the infectious-toxic symptoms that result in multiple organ infection and dysfunction. The isolates of inhaled and blood-borne SAP have different antibiograms.

Adolescent ; Age Factors ; Blood-Borne Pathogens ; isolation & purification ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Infant ; Male ; Pneumonia, Staphylococcal ; drug therapy ; microbiology ; Retrospective Studies