Pneumonia is the primary cause of death in children under five years of age.About 2 million children younger than 5 years die of pneumonia each year worldwide,accounting for 19％ of the total number of deaths.According to the survey in 2008,7％ ～ 13％ of the children with pneumonia are severe pneumonia.Severe pneumonia is a life-threatening disease which needs for intensive care.Cytokine IL-10 is a anti-inflammatory cytokine,and IL-18 is a proinflammatory cytokine,both of them are released by the inflammatory response,and can be involved in severe pneumonia immune-mediated.They are closely related to the occurrence,development and prognosis of severe pneumonia.

Objective:To optimize the water extraction technology of Xiaokeantangfang. Methods: The extraction technology of Xiaokeantangfang was optimized by orthogonal design with the content of astragaloside A and the extract yield as the indices and the a-mount of added water,extraction time and extraction times as the influencing factors,and the results were verified. Results:The amount of added water had significant effect on the content of astragaloside A(P<0. 05). The optimal extraction technology was as follows:12-fold amount of added water,extracting for 3 times with 0. 5 hour per time. The average extract yield was 38. 33%(RSD=1. 22%,n=3), and the average content of astragaloside A was 0. 0828 mg·g-1 for 3 batches of samples in the validation tests(RSD=3. 37%, n=3). Conclusion:The optimal water extraction technology of Xiaokeantangfang is stable and feasible, which can provide experimen-tal basis for the oral preparation of Xiaokeantangfang.

OBJECTIVE: Mutations in the GJB2 are the most common cause of nonsyndromic autosomal recessive sensorineural hearing loss. A few mutations in GJB2 have also been reported to cause dominant nonsyndromic or syndromic hearing loss. This study analysised the GJB2 dominant mutation in Chinese deafness. METHOD: 1641 patients as GJB2-related hearing loss were enrolled, summarized the type of dominant mutaion, analyzed the hearing level and other systerm lesion. RESULT: Nine probands with severe-profound hearing loss were diagnosed as GJB2 domiant mutation (R75W,G130V, R143Q,p. R184Q). And one patient with R75W mutation was diagosed as hearing loss and palmoplantar keratoderma. CONCLUSION GJB2 dominant mutation can cause severe-to-profound bilateral sensorineural hearing impairment and not common with syndromic hearing loss in Chinese deafness.
Asian Continental Ancestry Group ; Connexin 26 ; Connexins ; genetics ; Deafness ; genetics ; Hearing Loss ; Hearing Loss, Sensorineural ; Humans ; Keratoderma, Palmoplantar ; Mutation ; Phenotype

Background and purpose: Because of the aggressive nature of hilar cholangiocarcinoma and the absence of effective adjuvant therapy, surgical radical resection offers hilar cholangiocarcinoma patients the only choice. Research focus include preoperative assessment, the use of preoperative biliary drainage, the range of hepatic resection, and the range of lymphadenectomy. To investigate the clinical experience and efifcacy of combined hepatectomy in the treatment of hilar cholangiocarcinoma. Methods: Two hundred and seven patients with hilar cholangiocarcinoma treated surgically in the First Afifliated Hospital of Kunming Medical University form Jan. 2007 to Oct. 2013 were retrospectively analyzed. Results:Of the 207 patients, 125 patients who received radical resection (R0 resection) and the curative resection rate was 60.4%. One hundred and iffty-six cases were treated in combined hepatectomy group, 51 cases in non-hepatectomy group, the rate of R0 resection was 70.5%in hepatectomy group and 29.4%in non-hepatectomy group, and the difference was signiifcant (P<0.01). Two patients died perioperatively, the main postoperative complications included hepatic function insufifciency and bile leakage. One hundred and seventy-two patients were followed up, the median survival time of the 102 patients who received R0 resection was 45 months, and the 1, 3, 5 year survival rates were 96.1%, 59.1%and 17.2%. The median survival time of the 70 patients who received R1-2 resection was 26 months, and the 1, 3 year survival rates were 81.3%and 19.2%, and none of the patient survived for over 5 years. The survival rate of patients who received R0 resection was signiifcantly higher than those who received R1-2 resection (χ2=39.121, P<0.01). In the hepatectomy group was awarded the R0 resection in patients with postoperative 1, 3, 5 year survival rate was 97.8%, 63.9% and 18.0%, in non-hepatectomy group received R0 resection in patients with postoperative 1, 3, 5 year survival rate was 83.3%, 20.8%and 8.3%. There were signiifcant differences in the postoperative survival rate between both group (χ2=5.988, P=0.014). Conclusion:Radical excision is the key to improve the long term survival. Combined hemihepatectomy and standardized lymph node resection has signiifcantly improved the radical resection rate and the efifcacy of treatment for hilar cholangiocarcinoma.

Objective This study was performed to investigate the effect of hypoxia on glucose-6-phosphate isomerase (G6PI) expression and cell cycle of fibroblast-like synoviocytes from synovium of rheumatoid arthritis (RA) and osteoarthritis (OA) under hypoxia or normoxia.Methods Fibroblast-like synoviocytes were cultured with either of hypoxia (3％ oxygen) or normoxia (21％ oxygen) for 24 hours.The mRNA expression of G6PI and HIF-1α was tested by PCR quantification,while the protein levels of G6PI and HIF-1α were measured by western blot.Cell cycle was performed by FACS.T-test and Mann-Whitney U were used for statistical analysis.Results The expression levels of G6PI mRNA under hypoxia in RA were higher than those of OA (2.6±0.4 vs 1.5±0.4,P＜0.05).The protein levels of G6PI in RA were higher than those of OA (P＜0.05).The expression levels of HIF-1α mRNA under hypoxia in RA were higher than those of OA (2.9±0.8vs 1.4 ±0.4,P＜0.05).The protein levels of HIF-1α in RA were higher than those of OA (P＜0.05).The G1 phase ratio of cell cycle was decreased significantly under hypoxia than those of normoxia in RA ELs (t=1 1.31,P＜0.05).The S and G2 phase ratio of cell cycle were increased.Conclusion Hypoxia upregulates G6PI and HIF-1α expression and improves proliferation in fibroblast-like synoviocytes.

Objective To study the effect of autologous periosteum on the healing of tendon-bone interface in rabbit rotator cuff tear. Methods Supraspinatus tenotomy was performed on one side shoulder in 60 New Zealand white rabbits to establish the model of rotator cuff tear. The rabbits were randomly divided into two groups:the study group(used autologous periosteum to promote the suture fixation)and control group(simple suture fixation). The rabbits were sacrificed at 4,8 and 12 weeks postoperatively with twenty rabbits sacrificed each time. Tissue samples of the tendon-bone interface was taken for histological examination and biomechanical test was performed to assess the strength of tendon-bone junction. Results At 4 weeks the study group showed extensive inflammatory cell infiltration, and a small amount of chondrocytes and extracellular matrix. At 8 weeks, the study group showed a large amount of immature chondrocytes arranged rather regularly. At 12 weeks,the tendon bone junction appeared similar to normal. In the study group,both the tendon-bone connection and the arrangement of chondrocytes were significantly better than the control group. The result of biomechanical testing showed that the highest tendon load in the study group was significantly higher than the control group (P < 0.05). Conclusions The use of autologous periosteum as a patch to strengthen the repair of the rotator cuff tear can effectively promote the healing of tendon bone interface, shorten the rotator cuff healing time and has good biological properties. This method provides an experimental basis for clinical rotator cuff repair surgery.

OBJECTIVE: To analyze the curative effect of CI in children with GJB2-associated NSSNHL. METHOD: The evaluations of curative effect with CI include auditory threshold, IT-MAIS/MAIS, CAP, SIR. MESP. The outcomes of 40 cases with GJB2-associated NSSNHI, were compared 80 patients with negative results of screening of gene mutation (control group). RESULT: In comparison with control group the auditory threshold in children with GJB2-associated NSSNIL is better, however had no significant difference in other tests (P > 0.05). CONCLUSION CI could he performed on children with GJB2-associated NSSNHL. Postoperative outcomes of hearing and speech were satisfied.
Child ; Child, Preschool ; Cochlear Implantation ; Connexin 26 ; Connexins ; genetics ; Female ; Hearing Loss, Sensorineural ; genetics ; surgery ; Humans ; Infant ; Male ; Mutation ; Treatment Outcome

Objective To determine the audiological characteristics in 832 deaf children with biallelic causative mutations in GJB2 ,SLC26A4 gene .Methods The 832 patients received deafness gene screening ,553 were GJB2 gene biallelic causative mutations ,279 were SLC26A4 gene biallelic causative mutations .Patients were divided into four groups according to ages of hearing loss onset :<1 ,1~3 ,3~6 ,6~12 years old ,and the audiological character-istics and prevalence of GJB2 ,SLC26A4 gene mutations at different ages of onset .Results The prevalence of GJB2 gene mutations at four groups was 37 .97% (210/553) ,38 .34% (212/553) ,16 .27% (90/553) ,7 .41% (41/553) ,re-spectively ;the prevalence of SLC26A4 gene mutations at four groups was 25 .45% (71/279) ,44 .80% (125/279) , 20 .07% (56/279) ,9 .67% (27/279) ,respectively .The difference between GJB2 and SLC26A4 gene was significant(P=0 .001) .The prevalence of profound hearing loss with GJB2 gene mutations at four groups were 66 .67% (140/210) ,61 .32% (130/212) ,47 .78% (43/90) ,41 .46% (17/41) ,respectively .The difference was significant (P=0 .004) ,while the difference in 279 patients with SLC26A4 gene mutations was not statistically significant (P= 0 . 083) .Conclusion The age of hearing loss onset in patients with biallelic causative mutations in GJB 2 or SLC26A4 gene refers to 0~3 years -old ,hearing loss in patients with GJB2 ,SLC26A4 gene mutations gives priority to pro-found .The age of hearing loss onset is smaller ,the ratio of profound hearing loss is higher .Patients with severe and profound hearing impairment should be performed the genetic testing when the age of onset under 12 .

BACKGROUND:A large number of studies have confirmed that exosomes can promote osteogenesis and vascularization.However,simple exosome therapy has problems such as poor targeting,and the content of loaded molecules cannot reach the therapeutic concentration. OBJECTIVE:To load exosomes into injectable gluconolactone-sodium alginate β-tricalcium phosphate-polyethylene glycol hydrogel,and observe the effect of the hydrogel on peri-implant bone defect in vivo and in vitro. METHODS:Exosomes were extracted from bone marrow mesenchymal stem cells and wrapped in injectable gluconolactone-sodium alginate β-tricalcium phosphate-polyethylene glycol hydrogel.(1)In vitro experiment:The hydrogel loaded with exosomes and the hydrogel without exosomes were cocultured with endothelial progenitor cells,and exosomes uptake experiment,tubule formation experiment,cell proliferation,migration ability,and angiogenic gene detection were carried out.(2)In vivo experiment:Twelve male New Zealand white rabbits were used to prepare two standard implant cavities and corresponding bone defects in the long axis of one femur.A hydrogel loaded with exosomes was implanted in the bone defect after an implant was implanted in a cavity at the proximal end of the implant(experimental group),and an unloaded exosome hydrogel was implanted in the bone defect after an implant was implanted in a cavity at the distal end of the implant(control group).At 3,6 and 9 weeks after operation,bone defects with implants were removed and stained with hematoxylin-eosin staining and Masson staining.Simultaneously,osteogenic and angiogenic genes were detected at 9 weeks after operation. RESULTS AND CONCLUSION:(1)In vitro experiment:Exosomes could enter endothelial progenitor cells.The proliferation,migration,angiogenesis and gene(CD31,vascular endothelial growth factor and basic fibroblast growth factor)expression of endothelial progenitor cells in the hydrogel-loaded group were higher than those in the hydrogel-unloaded group(P<0.05).(2)In vivo experiment:Hematoxylin-eosin staining and Masson staining showed that at 3 weeks after operation,only a small amount of new bone was found in the two groups,and the material was partially degraded.At 6 weeks after operation,the amount of new bone in the two groups increased,and a large amount of new bone was found in the experimental group,with obvious calcium deposition.At 9 weeks after operation,compared with the control group,a large number of bone trabeculae thicker than mature were found in the experimental group,calcium salt deposition was more obvious,and a large number of osteoblasts were found around the bone trabeculae.The protein expressions of CD31,vascular endothelial growth factor,basic fibroblast growth factor,bone morphogenetic protein 2,type I collagen and osteocalcin in the experimental group were higher than those in the control group at 9 weeks after operation(P<0.05).(3)The exosome-loaded gluconolactone-sodium alginate β-tricalcium phosphate-polyethylene glycol hydrogel could promote the proliferation,migration and angiogenic differentiation of endothelial progenitor cells and promote the repair and regeneration of bone defects around implants.

OBJECTIVE: To analyze the positive rate of common genetic mutations in Chinese non-syndromic sensorineural hearing loss groups with different hearing phenotype. METHOD: One thousand four hundred and forty-eight subjects with hearing test results received at least one of three genetic testings including: mutations in coding region of GJB2 and SLC26A4 with sequencing analysis and mitochondrial DNA C1494T/A1555G with microarray detection. Of 1448 subjects, 1333 have bilateral sensorineural hearing loss, 65 have unilateral hearing loss and 50 have normal hearing threshold even though they have high frequency hearing loss or family history. The informed consent of each subject was achieved. RESULT: Mutation positive rate of GJB2, SLC26A4 and mtDNA C1494T/ A1555G of 1448 subjects were 19.23%, 27.55%, 0.1% and 1.72% respectively. The positive rate of GJB2 and SLC26A4 mutations in bilateral hearing loss group (20.22%, 29.17%) was statistically significantly higher than unilateral group (0, 0) (P < 0.01). In bilateral hearing loss group, the positive rate of GJB2 mutations was highest in the profound group (24.67%), and then severe (22.33%), moderate (14.33%) and mild group (6.58%) (P < 0.01). The positive rate of SLC26A4 mutations was highest in the severe group (48.67%), and then profound (28.42%), moderate (21.16%) and mild (8.93%) (P < 0.01). CONCLUSION The positive rate of GJB2 and SLC26A4 mutations is high in the groups with bilateral profound and severe sensorineural hearing loss, whose genetic testing should be put emphasis on. However, the genetic testing should be performed in patients with mild to moderate hearing impairment as well if necessary.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Audiometry ; Child ; Child, Preschool ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; DNA, Mitochondrial ; genetics ; Female ; Hearing Loss ; genetics ; Hearing Loss, Bilateral ; genetics ; Hearing Loss, High-Frequency ; genetics ; Hearing Loss, Sensorineural ; genetics ; Hearing Loss, Unilateral ; genetics ; Humans ; Infant ; Male ; Membrane Transport Proteins ; genetics ; Middle Aged ; Mutation ; Phenotype ; Sulfate Transporters ; Young Adult