Objective To investigate the characteristics of traditional Chinese medical syndrome types of fulminant and epidemic dengue fever patients admitted in Guangzhou and Xishuang banna in the year of 2013,and to ex plore the differences of etiology and pathogenesis, illness, and treatment for the patients in the two regions. Methods We collected the clinical data of 78 cases receiving integrative Chinese and western medicine from 255 patients admitted in Guangzhou Municipal Eighth People’s Hospital, and the clinical data of 39 cases receiving integrative Chinese and western medicine from 120 patients admitted in the People’s Hospital of Xishuangbanna Dai Nationality Autonomous Prefecture in the year of 2013. The traditional Chinese medical syndrome types and the syndrome scores of the total of 117 cases were investigated. The method of phenology was used for the analysis of epidemic time and epidemic region of dengue fever, and the theory of defense-qi-nutrient-blood syndrome differentiation of seasonal febrile diseases was used for the analysis of etiology and pathogenesis of dengue fever. Results ( 1) Dengue fever was epidemic in the first ten days of July and in the middle ten days of November of the year 2013 in Guangzhou region, and was epidemic in the middle ten days of August and the first ten days of October in Xishuangbanna region. The epidemicity of dengue fever in Guangzhou covered the end of summer and the whole autumn, and then disappeared before the coming of winter. In Xishuangbanna , the epidemicity of dengue fever was obvious in autumn, and disappeared in late autumn. ( 2) In the two hospitals, dengue fever patients were dominated by the syndromes of excessive heat in both Qifen and Xuefen, blood stasis blended with toxicity, excessive heat in Qifen, and lingering pathogens in order. (3) Before treatment, the scores of fever were higher in patients of Xishuangbanna hospital than those in patients of Guangzhou Eighth People’s Hospital ( P<0.01) . After treatment for 3 days, fever scores were improved in both hospitals (P<0.05 or P<0.01), but the differences between the two hospitals were insignificant (P>0.05) . After treatment for 6 days, fever disappeared in patients of both hospitals. (4) Before treatment, the scores of syndromes were higher in patients of Xishuangbanna hospital than those in patients of Guangzhou Eighth People’s Hospital ( P<0.05) . After treatment for 3 days, syndorme scores were improved in both hospitals ( P<0.01) , but the syndrome scores were higher in Xishuangbanna hospital than those in patients of Guangzhou Eighth People’s Hospital. After treatment for 6 days, syndrome scores were much improved in patients of both hospitals compared with those after treatment for 3 days (P<0.01) . Conclusion In dengue fever patients admitted in Guangzhou and Xishuangbanna region, the syndrome of excessive heat in both Qifen and Xuefen is the leading type, and then comes blood stasis blended with toxicity. The illness state of patients in Guangzhou region is milder than that of the patients in Xishuangbanna region, the time for symptom relief is about one week, and similar therapeutic effect can be achieved in the two regions .

This study was aimed to construct eukaryotic expression vectors carrying the small hairpin RNA (shRNA) targeting TRPC6 gene and investigate the effect of TRPC6 knockdown on puromucin aminonucleoside (PAN)-induced podocyte injury. Two DNA sequences containing the small hairpin structure targeting TRPC6 were designed, synthesized and then inserted into the green fluorescence protein (GFP)-contained plasmids (pGC) to establish the plasmids pGCsi-TRPC6A and pGCsi-TRPC6B. Plasmids expressing scrambled shRNA were used as negative control and named pGCsi-NC. These plasmids were transfected into a conditionally immortalized murine podocyte cell line by using liposome. Flow cytometry was used to examine the transfection efficiency. TRPC6 mRNA and protein expression levels were detected by RT-PCR and Western blotting. Cultured podocytes were divided into four groups: control group, PAN treatment group, PAN+TRPC6 shRNA transfected group and PAN+scrambled shRNA transfected group. The paracelluar permeability to BSA was evaluated by Millicell-PCF Inserts and cell viability was measured by the trypan blue assay. Immunofluorescent assay was used to observe the distribution of α-actinin-4 and α-tubulin. The results showed that the transfection efficiency of the shRNA expression vector was about 45%. Expression levels of TRPC6 mRNA and protein were downregulated after transfection with pGCsi-TRPC6A and pGCsi-TRPC6B. Knocking down TRPC6 gene could effectively reverse the PAN-induced increase in the paracelluar permeability to BSA. The distribution of α-actinin-4 and α-tubulin was disrupted after treatment with PAN, which was reversed by knocking down TRPC6 gene. It was concluded that knocking down TRPC6 gene could effectively prevent podocytes from the permeability increase induced by PAN, which may be related to the regulation of podocyte cytoskeleton.

This study was aimed to construct eukaryotic expression vectors carrying the small hairpin RNA (shRNA) targeting TRPC6 gene and investigate the effect of TRPC6 knockdown on puromucin aminonucleoside (PAN)-induced podocyte injury. Two DNA sequences containing the small hairpin structure targeting TRPC6 were designed, synthesized and then inserted into the green fluorescence protein (GFP)-contained plasmids (pGC) to establish the plasmids pGCsi-TRPC6A and pGCsi-TRPC6B. Plasmids expressing scrambled shRNA were used as negative control and named pGCsi-NC. These plasmids were transfected into a conditionally immortalized murine podocyte cell line by using liposome. Flow cytometry was used to examine the transfection efficiency. TRPC6 mRNA and protein expression levels were detected by RT-PCR and Western blotting. Cultured podocytes were divided into four groups: control group, PAN treatment group, PAN+TRPC6 shRNA transfected group and PAN+scrambled shRNA transfected group. The paracelluar permeability to BSA was evaluated by Millicell-PCF Inserts and cell viability was measured by the trypan blue assay. Immunofluorescent assay was used to observe the distribution of α-actinin-4 and α-tubulin. The results showed that the transfection efficiency of the shRNA expression vector was about 45%. Expression levels of TRPC6 mRNA and protein were downregulated after transfection with pGCsi-TRPC6A and pGCsi-TRPC6B. Knocking down TRPC6 gene could effectively reverse the PAN-induced increase in the paracelluar permeability to BSA. The distribution of α-actinin-4 and α-tubulin was disrupted after treatment with PAN, which was reversed by knocking down TRPC6 gene. It was concluded that knocking down TRPC6 gene could effectively prevent podocytes from the permeability increase induced by PAN, which may be related to the regulation of podocyte cytoskeleton.
Animals ; Cell Membrane Permeability ; drug effects ; physiology ; Cell Survival ; drug effects ; physiology ; Cells, Cultured ; Gene Knockdown Techniques ; Mice ; Mice, Knockout ; Podocytes ; drug effects ; physiology ; Puromycin Aminonucleoside ; pharmacology ; TRPC Cation Channels ; genetics ; metabolism