Objective To investigate the effects of Hui-hui Gan-song Yin(HGY) on the accumulation of extracellular matrix of rat glomerular mesangial cells(MCs) induced by high glucose.Methods The 40 SD rats were randomly divided into normal control group(distilled water), glurenorm group(10 mg/kg), HGY high-dose group(10 g/kg) and HGY low-dose group(5 g/kg), 10 rats in each group.The rats in each group were treated with corresponding drugs, twice a day.After 3 days, the serum containing each drug were prepared to culture rat MCs in vitro.The MCs were divided into the normal control group( 10% serum of rats in normal control group ) , high glucose group ( 30 mmol/L glucose +10% serum of rats in normal control group), glurenorm group(30 mmol/L glucose+10% serum of rats in glurenorm group), HGY high-dose group(30 mmol/L glucose+10% serum of rats in HGY high-dose group) and HGY low-dose group(30 mmol/L glucose+10% serum of rats in HGY low-dose group).The fibronectin(FN), ColⅠand ColⅣ levels were detected by Western blot.Results Compared with normal control group, the expression of FN, ColⅠand ColⅣ in high glucose group increased(P<0.01).The HCY suppressed the protein expression of FN, ColⅠand ColⅣ significantly(P<0.05).Conclusion The serum containing HGY could suppressed protein expression of FN , ColⅠand ColⅣ and inhibit the accumulation of extracellular matrix of MCs induced by high glucose, which could protect glomerulus and delay the development of diabetic nephropathy.

Objective To evaluate the expression characteristics of calcium-binding protein S100A8 (Calgranulin A) and S100A9 (Calgranulin B) in lung squamous cell carcinoma and lung adenocarcinoma and their correlations with clinical data.Methods Twenty paired snap frozen cancer tissue and para-cancer tissue of lung cancer patients were collected form Jan 2013 to Dec 2014 in our hospital.And other 100 cancer paraffin wax specimen,20 para-cancer paraffin wax specimen were retrospectively collected from Jan 2007 to Dec 2009.The expressions of S100A8 and S100A9 were analyzed by PCR,Western blot and immunohistochemistry assay.The association between S100A8,S100A9 expression and patients clinical characteristics were analyzed.Results Positive rates of S100A8,S100A9 mRNA were 100.0％ for cancer tissue,significant higher than those of para-cancer tissue of 40.0％ and 85.0％ (P ＜0.05).Positive rates of S100A8,S100A9 protein were 100.0％ for cancer tissue,significant higher than those of para-cancer tissue of 20.0％ and 90.0％ (P ＜ 0.05).S100A8 positive expression rate was 28.6％ and 89.5％ for well differentiation and poor differentiation groups with statistical difference (P ＜ 0.05).The expression rate of S100A9 for stage Ⅰ positive was 73.0％ and the 100.0％ of S100A9 for stage Ⅲ/Ⅳ groups with statistical difference (P ＜ 0.05).Conclusion The expression levels of S100A8 and S100A9 in lung adenocarcinoma and lung squamous cell carcinona was higher than those in normal tissue,which were negatively correlated with the degree of differentiation of lung adenocarcinoma,and without correlation to the degree of differentiation of lung squamous cell carcinoma.The expression level of S100A9 was negatively correlated with tumor TNM stage.

Hepatitis B is one of the most serious global threats to human health. Phylogenetic analysis of hepatitis B virus (HBV) can reveal the evolutionary relationship between HBV sequences and thus provide a basis for the prediction and treatment of hepatitis B and other aspects. In this study, we performed sequence analyses on the HBV sequences of five clinical HBV samples and the HBV sequences retrieved from the GenBank, EMBL, and DDBJ to construct a phylogenetic tree and analyze sequence structures. The experimental results revealed that the C gene of one cloned sequence had a recombinant structure of HBV B/ C subtype. Moreover, the phylogenetic results proved the existence of a newly found subtype HBV/B6 in Xishuangbanna of Yunnan Province, China. The experimental conclusion represents certain value for phylogenetic studies of HBV in Yunnan ethnic minority groups.
DNA, Recombinant ; genetics ; Genes, Viral ; genetics ; Genotyping Techniques ; Hepatitis B virus ; classification ; genetics ; Humans ; Phylogeny

Objective To investigate the effect of progressive neurological deterioration ( PND) of cerebral watershed infarction on early prognosis. Methods The consecutive patients with cerebral watershed infarction admitted in the Department of Neurology,Jinling Hospital,Nanjing University School of Medicine and their cerebral watershed infarctions confirmed by the imaging examination from March 2009 to March 2014 were enrolled. The clinical features, laboratory indicators and imaging features of internal watershed infarction,cortical-type watershed infarction,and mixed watershed infarction were identified and analyzed. The National Institutes of Health Stroke Scale was used to score neurological deficit. The modified Rankin scale ( mRS) was used to score the prognosis of patients. Single factor analysis was used to compare the differences between the groups. At the same time,the correlation between PND and poor prognosis of cerebral watershed infarction at day 90 was analyzed by multivariable Logistic regression analysis. Results A total of 89 patients with cerebral watershed infarction were enrolled,including 43 cortical-type watershed infarctions,36 internal watershed infarctions, and 10 mixed watershed infarctions. Single factor analysis indicated that the incidences of PND of internal watershed infarction and mixed watershed infarction were significantly higher than the cortical-type watershed infarction (36. 1% [n=13],50. 0% [n=5], and 16. 3% [n=7],respectively;P=0. 018). At day 90,28 patients had poor prognosis,and mRS was (3.4±1. 0) scores at day 90. There was significant difference in the types of infarction between the patients with poor prognosis and patients with good prognosis (P<0. 05). In patients with poor prognosis, most of them were internal watershed infarctions,accounting for 50. 0% (14/28),while in patients with good prognosis,most of them were cortical-type watershed infarctions(57. 4% [35/61]). The incidence of PND in patients with poor prognosis was significantly higher than that in patients with good prognosis (57.1% [16/28] vs. 14. 8% [9/61];P<0. 05). The result of multivariate Logistic regression analysis showed that after adjustment for confounding factor, PND was independently associated with the poor prognosis of cerebral watershed infarction at day 90 (OR 6. 969,95%CI 2. 451-19. 869;P<0. 01). Conclusion Compared with the cortical-type watershed infarction, the patients with internal watershed infarction is more prone to have PND, and PND is independently correlate with the poor prognosis at day 90.

Objective To study the relationship of proliferation and activation of T lymphocyte subsets and disease progression in antiretroviral-naive human immunodeficiency virus(HIV)-1-infected individuals.Methods Forty-nine antiretroviral-naive,chronically HIV-1 infected patients and 16 healthy,HIV-1 negative controls were enrolled in this study.The patients were divided into 3 groups according to their CD4+T cell counts:<200×106/L,(200-350)×106/L and>350×106/L.Peripheral blood mononuclear cells(PBMC)were isolated.T cell proliferation index was measured by Ki-67 staining.T cell activation was detected by CD38 staining.The samples were analyzed by flow cytometry.The data were compared by one-way ANOVA.Results The percentage of Ki-67+cells in CIM+T ceils was 7.92%±4.37%in CD4+T cell<200×106/L group,which was significantly higher than those 0.39%d:0.24%in control group,2.61%±2.12%in(200-350)×106/k group and 2.65%±2.13%in>350 X106/L group(F=21.961,P<0.01).The percentage of Ki-67+cells in CD8+T ceils in CD4+T cells<200×106/L group was 2.87%±1.13%,which was also much higher than those in other 3 groups(0.15%±0.90%,1.40%±1.17%,1.22%±0.80%,respectively F=19.203,P<0.01).The Ki-67'CD4'T cells and Ki-67+CD8+T cells were inversely correlated with CD4+T cell counts(r=-0.654,r=-0.539,respectively;P350×106/L group(F=14.333,F=15.412,respectively;P<0.01).The expressions of Ki-67+ on CD4+ and CD8+T cells were positively correlated with CD38 expression(r=0.527,r=0.391,respectively;P=0.002).Conclusions The proliferation and activation of T lymphoeytes are enhanced during HIV/AIDS disease progression.And T eell activation may be the result of persistent immune activation.

Objective To evaluate the feasibility and safety profile of pegylated-interferonα-2a (Peg IFNα-2a) combined with adefovir dipivoxil (ADV) in inactive hepatitis B surface antigen (HBsAg) carriers (IHC).Methods This was a single center, prospective and open-label study.IHC were divided into therapeutic group (T, 112 subjects) and control group (C, 72 subjects) according to personal willingness.Patients with hepatitis B virus (HBV) DNA<20 IU/mL were treated with Peg IFNα-2a monotherapy, and those with HBV DNA ≥20-<2 000 IU/mL were treated with Peg IFNα-2a combined with ADV.Total therapy duration was 96 weeks.For patients who achieved HBsAg seroconversion and continued consolidation treatment for 24 weeks, the treatment duration could be less than 96 weeks.t test was used for continuous variable comparison between the two groups, while chi-square test or Fisher′s exact probability method was used for counting data analysis.The related factors affecting HBsAg clearance was analyzed by univariate or multivariate logistic regression analysis.Results A total of 194 patients were enrolled with 112 in therapeutic group and 72 in control group.The HBsAg clearance rate and seroconversion rate at week 48 in therapeutic group were 30.8% (32/104) and 26.0% (27/104), respectively.The rates at week 96 increased to 45.2% (47/104) and 38.5% (40/104), respectively.The HBsAg clearance rates at weeks 48 and 96 in control group were both 1.5% (1/68).HBsAg seroconversion was not achieved in control group.The HBsAg clearance rate in treatment group was significantly higher than that in control group (χ2=39.066, P<0.01).The quantitative HBsAg levels at baseline (OR=2.313, 95%CI: 1.258-4.251, P=0.007), week 12 (OR=3.159, 95%CI: 1.826-5.466, P<0.01) and week 24 (OR=3.347, 95%CI: 2.050-5.465, P<0.01), the decline of HBsAg at week 12 (OR=5.343, 95%CI: 2.085-13.689, P<0.01), and week 24 (OR=4.855, 95%CI: 2.380-9.902, P<0.01), and alanine transaminase (ALT) elevation at week 12 (OR=3.520, 95%CI: 1.369-9.052, P=0.009) were independent predictors for HBsAg clearance.Conclusions Peg IFNα-2a-based treatment for IHC could achieve higher HBsAg clearance rate and seroconversion rate, and has a safety profile.Decline of HBsAg at week 12 and week 24 with ALT elevation at week 12 could predict a higher HBsAg clearance rate.

Objective To explore the effects of amniotic lacrimal duct stenting on the prevention of dry eye in castrated rabbits.Methods Thirtysix healthy male rabbits were selected,the third eyelid were cut off and antiinfection treatment were given,which were randomly divided into 3 groups (12 cases in each group),the castrated male rabbits models were made.Among them,group A was negative control group,group B was dry eye model group,group C was group of lacrimal amniotic membrane group.At 2 weeks before implantation of amniotic lacrimal duct stent,2 weeks,4 weeks and 6 weeks after implantation,the fluorescent (FL) examination,Western blot,Schirmer I examination,immunofluorescence staining and corneal confocal microscopy were performed.Results The levels of tear secretion and FL in the three groups among different time points were significantly different (F=7.126,P =0.009;F =9.658,P =0.016),and there were significant differences among three groups (F =12.582,P =0.005;F =13.187,P =0.013).The tendency of tear secretion and FL in the three groups were also significantly changed (F =8.531,P =0.007;F =10.652,P =0.019).The epithelial basal cells at 6 weeks after implantation in three groups were 3811 ±414,3820 ± 314,2789 ± 353,and the density of inflammatory cells was 266 ±28,266 ± 29,67 ± 13,there were significant differences among three groups (F =13.442,P =0.012;F =9.231,P =0.021).The K1 6 staining in the duct epithelium were negative,and the expression of α-SMA in the lacrimal duct tissue of group A,B and C was not changed at all time points after implantation of amniotic lacrimal stent,and there was no significant difference (F =14.681,P =0.002).Conclusion The amniotic lacrimal stent implantation has certain effect on the prevention of dry eye in rabbit.

OBJECTIVETo explore effects of 650 nm laser and moxibustion pretreatment on visceral traction pain (VTP) and its mechanism.

METHODSForty male SD rats were randomly devided into a sham operation group (group A), a VTP group (group B), a 650 nm laser pretreatment group (group C), a moxibustion pretreatment group (group D). Rats in group A and group B were not treated except sham operation or VTP model. In group C and D, the VTP models were produced immediate after 650 nm laser irradiation or moxibustion at "Zusanli" (ST 36), respectively. The changes of pain score and systolic pressure were investigated and the activity of AChE, the content of SP and leu-enkephaline (LEK), and the positive index of c-Fos protein and glial fibrillary acidic protein (GFAP) were detected by biochemistry, radio-immunity method and immunohistochemistry, respectively.

RESULTSCompared with group A, the pain score, systolic pressure, the activity of AChE, the content of SP, and the positive index of c-Fos protein and GFAP of group B increased significantly (all P < 0.05); compared with group B, the pain score, AChE activity, the content of SP and the positive index of c-Fos protein and GFAP of both group C and group D decreased significantly (all P < 0.05); compared with group B, the content of LEK increased and systolic pressure decreased significantly in group C (both P < 0.05).

CONCLUSIONBoth 650 nm laser and moxibustion pretreatment can inhibit VTP and the mechanism may be related to reducing the activity of AChE and the content of SP, and increasing the activity of LEK and decreasing the expression of c-Fos protein and GFAP.

Animals ; Combined Modality Therapy ; Enteric Nervous System ; physiopathology ; Humans ; Intestinal Pseudo-Obstruction ; physiopathology ; therapy ; Laser Therapy ; Male ; Moxibustion ; Pain ; physiopathology ; Pain Management ; Random Allocation ; Rats ; Rats, Sprague-Dawley

To study the dynamic change law of bioactive constituents from Polygonum multiflorum, and to explore the optimal harvest period of P. multiflorum. Determination of stilhene glucoside, anthraquinones and catechin from P. multiflorum in different harvest times by MEKC-DAD, and principal component analysis (PCA) was used to comprehensive evaluation for bioactive constituents. There are obvious differences among the contents of active ingredients in various collecting periods samples, the content of stilbene glucoside was the highest in November, the total content of combined anthraquinone was the highest in November and December, the content of catechin was the highest in September. The comprehensive evaluation index obtained with principal component analysis showed that the sample collected in November is significantly higher than those with other samples. The optimal harvest period of P. multiflorum is November.
Electrophoresis ; Fallopia multiflora ; chemistry ; growth & development ; metabolism ; Time Factors

OBJECTIVETo delineate the characteristics of the clinical manifestation, pathology of skeletal muscle and gene mutations of mitochondrial encephalomyopathy with lactic acidosis and stroke-like episode (MELAS) in children.

METHODThe clinical manifestation, laboratorial data, brain images, muscle pathology and mitochondrial gene mutations were analyzed in 24 patients with MELAS who were diagnosed in Department of Pediatrics, Peking University First Hospital. Their prognosis was evaluated by following up.

RESULTSymptoms of central nervous system such as stroke-like episodes, vomiting, convulsion and headache were present in all the 24 cases. Nine cases had the symptoms of myopathy. Twenty cases had developmental delay. Short stature, being thin and hairy was very common in these cases. Serum lactate level increased in all the cases, pyruvate increased in 17 cases. Elevated CSF lactate was found in 2 cases. Magnetic resonance imaging (MRI) was performed on 24 cases, out of them 23 were abnormal. The lesions mainly involved cerebral lobes. Occipital lobe was the most common site of lesions. Computed tomography (CT) was performed on 13 cases, low density lesions were present in 10 cases, basal ganglia calcifications in 5 cases. Muscle biopsy was performed on 8 cases, ragged-red fibers (RRF) were found in 4/8 cases, and abnormal accumulation of mitochondria were found in 3/8 cases. The mtDNA gene mutational analysis showed A3243G mutation in these patients. The mutation rates varied from 11.6% to 75.0%. The same mutation were found in 4/5 mothers who had the genetic tests, and the mutation rates of the mothers varied from 15.0% to 23.6%. The clinical information of 11 cases was available through recent following up. Three cases died, the others had some degrees of mental retardation.

CONCLUSIONChildren with MELAS had various clinical manifestations. Central nervous system and skeletal muscle were usually involved. Short stature and hypertrichosis were common signs. The prognosis of this disease was disappointing. mtDNA A3243G was the most common mutation in MELAS. Fully understanding the characteristics of its clinical manifestation, laboratory tests, brain image, muscle pathology and molecular features can be helpful to the early diagnosis and treatment.

Acidosis, Lactic ; blood ; Adolescent ; Brain ; diagnostic imaging ; pathology ; Child ; Child, Preschool ; DNA Mutational Analysis ; DNA, Mitochondrial ; genetics ; Electroencephalography ; Female ; Follow-Up Studies ; Humans ; Infant ; MELAS Syndrome ; diagnosis ; genetics ; pathology ; Magnetic Resonance Imaging ; Male ; Muscle, Skeletal ; diagnostic imaging ; pathology ; Point Mutation ; Pyruvic Acid ; blood ; Stroke ; diagnostic imaging ; genetics ; pathology ; Syndrome ; Tomography, X-Ray Computed