Effect and mechanism of Puerariae Lobatae Radix in alleviating insulin resistance in T2DM db/db mice based on intestinal flora / 中国中药杂志

This study aimed to examine the effect and underlying mechanism of Puerariae Lobatae Radix on insulin resistance in db/db mice with type 2 diabetes mellitus(T2DM) based on the analysis of intestinal flora. Fifty db/db mice were randomly divided into a model group(M group), a metformin group(YX group), a high-dose Puerariae Lobatae Radix group(YGG group), a medium-dose Puerariae Lobatae Radix group(YGZ group), and a low-dose Puerariae Lobatae Radix group(YGD group). Another 10 db/m mice were assigned to the normal group(K group). After continuous administration for eight weeks, body weight and blood sugar of mice were measured. Enzyme linked immunosorbent assay(ELISA) was used to detect glycosylated serum protein(GSP) and fasting serum insulin(FINS), and insulin resistance index(HOMA-IR) was calculated. The histopathological changes in the pancreas were observed by HE staining. Tumor necrosis factor(TNF)-α expression in the pancreas was detected using immunohistochemistry. The structural changes in fecal intestinal flora in the K, M, and YGZ groups were detected by 16S rRNA. Western blot was used to detect the expression of farnesoid X receptor(FXR) and takeda G protein-coupled receptor 5(TGR5) in the ileum, cholesterol 7α-hydroxylase(CYP7A1) and sterol 27α-hydroxylase(CYP27A1) in the liver, and G protein-coupled receptors 41(GPR41) and 43(GPR43) in the colon. Compared with the K group, the M group showed increased body weight, blood sugar, serum GSP, fasting blood glucose(FBG), and FINS, increased HOMA-IR, inflammatory infiltration of islet cells, necrosis and degeneration of massive acinar cells, unclear boundary between islet cells and acinar cells, disturbed intestinal flora, and down-regulated FXR, TGR5, CYP7A1, CYP27A1, GPR41, and GPR43. Compared with the M group, the YX, YGG, YGZ, and YGD groups showed decreased body weight, blood sugar, serum GSP, FBG, and FINS, islet cells with intact and clumpy morphology and clear boundary, necrosis of a few acinar cells, and more visible islet cells. The intestinal flora in the YGZ group changed from phylum to genus levels, and the relative abundance of intestinal flora affecting the metabolites of intestinal flora increased. The protein expression of FXR, TGR5, CYP7A1, CYP27A1, GPR41, and GPR43 increased. The results show that Puerariae Lobatae Radix can improve the inflammatory damage of pancreatic islet cells and reduce insulin resistance in db/db mice with T2DM. The mechanism of action may be related to the increase in the abundance of Actinobacteria, Bifidobacterium, and Bacteroides in the intestinal tract and the protein expression related to metabolites of intestinal flora.

Relationship between vitamin D deficiency and necrotizing enterocolitis in preterm infants / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To evaluate the relationship of vitamin D level with the development of necrotizing enterocolitis (NEC) in preterm infants.</p><p><b>METHODS</b>A total of 429 preterm infants with a gestational age of <36 weeks, who were admitted to the department of neonatology within 2 hours after birth between January and December, 2016, were enrolled in the study. According to whether these infants developed NEC, the 429 subjects were divided into NEC group (n=22) and non-NEC group (n=407). Peripheral venous blood was collected from these preterm infants and their mothers at admission to measure the level of 25-hydroxyvitamin D (25-OHD). The two groups were compared in terms of the serum 25-OHD levels of preterm infants and their mothers. Pearson correlation analysis was used to investigate the correlation between the serum 25-OHD levels of preterm infants and their mothers. The distribution of vitamin D levels in preterm infants was compared between the two groups. The univariate logistic regression analysis was used to determine the risk factors for NEC in preterm infants.</p><p><b>RESULTS</b>The serum 25-OHD levels of preterm infants and their mothers in the NEC group were significantly lower than in the non-NEC group (P<0.001). In both groups, the serum 25-OHD levels of mothers and preterm infants were positively correlated with each other (P<0.001). The distribution of vitamin D levels (normal vitamin D level, low vitamin D level, vitamin D deficiency, and severe vitamin D deficiency) was significantly different between the NEC and non-NEC groups (P<0.001). The univariate logistic regression analysis showed that gestational age, birth weight, 25-OHD levels of preterm infants and their mothers, the duration of mechanical ventilation, the duration of oxygen inhalation, and the length of hospital stay were associated with the development of NEC (P<0.05).</p><p><b>CONCLUSIONS</b>The serum 25-OHD levels of preterm infants and their mothers may be related to the development of NEC in preterm infants, suggesting that vitamin D supplementation during pregnancy is important for preventing the development of NEC in preterm infants.</p>

Relationship between vitamin D deficiency and early-onset neonatal sepsis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To evaluate the effect of vitamin D level on early-onset sepsis (EOS) in neonates.</p><p><b>METHODS</b>Seventy-eight full-term neonates with EOS were used as the research group (EOS group). sixty healthy full-term neonates without clinical and/or laboratory features related to infections were used as the control group. Blood samples of the neonates and their mothers in both groups were collected within 72 hours of delivery to determine 25-hydroxyvitamin D(25-OHD) levels. The rate of vitamin D deficiency in the neonates and the level of 25-OHD supplemented to their mothers during pregnancy were compared between the two groups.</p><p><b>RESULTS</b>There was a significant positive correlation between the serum level of 25-OHD of the mothers and that of the neonates in both groups (EOS group: r=0.797, P<0.01; control group: r=0.929, P<0.01). The neonates and their mothers in the EOS group had significantly lower 25-OHD levels than those in the control group (P<0.01). The rate of vitamin D deficiency among the neonates in the EOS group was significantly higher than that of the control group (P<0.01). The level of vitamin D supplemented to the mothers during the last 3 months of pregnancy in the EOS group was significantly lower than that in the control group (P<0.01).</p><p><b>CONCLUSIONS</b>Low serum level of 25-OHD is associated with the development of early-onset sepsis in full-term neonates.</p>

Screening of biological amniotic immunization safety / 国际药学研究杂志

Objective: To evaluate the immunity efficacy of human amniotic membranes on rats. Methods: One hundred and fifty Wistar rats were randomly divided into five groups: biological amnion group, immunosuppression group, immunostimulation group, sham-operated group and blank control group. According to the study period, each group of thirty rats would be randomly divided into five experimental operation subgroups: the 1st week, the 2nd week, the 4th week, the 8th week group and the 12th week groups. The rats were implanted subcutaneously, then intramuscular injection of gentamicin sulfate for 3 days to resist the infection, and the immune organ coefficient, and the killing abilities of NK cell, IL-1β, IL-6 and TNF-αserum levels were detected according to the study period. Results: At 1st, 2nd, 4th, 8th and 12th week after amniotic membrane implantation in rats, compared with the sham-operated and blank control groups, the biological amnion group had nonsignificant differences(P>0.05). At 1st week after amniotic membrane implantation in rats, immunosuppression group showed different levels of the immunosuppressive effect, such as the analysis of immune organ coefficient, which had significant differences compared with other groups(P<0.01). At 1st week after amniotic membrane implantation in rats, the imunostimulation group showed a certain degree of the immunostimulant effect, such as the killing abilities of NK cell, which had marked differences compared with other groups(P<0.05). Conclusion: The amniotic membranes have satisfactory immune safety with implantation in rats and do not cause significant adverse immune reactions.

Long-term results of postoperative electronic irradiation for 53 patients with keloids / 中华整形外科杂志

<p><b>OBJECTIVE</b>To analyze the results of postoperative radiotherapy with electronic beam for patients with keloids in our hospital.</p><p><b>METHODS</b>From September 2006 to May 2009, radiotherapy was given within 24 hours after operation in 53 keloid patients. With single vertical field irradiation, 6-12 Mev electronic beams of Linear Accelerator were selected for different incision depth in different sites. The field size was 1.0 cm (range: 0.5-2.0 cm) away from both incision ends and 1.25 cm (range: 0.75-2.50 cm) away from incision laterally. The radiation was given daily with median treatment course of 4 days (range: 3-21 days) at 3.5 Gy/Fx to a median total dose of 14 Gy (range: 8-20 Gy). SPSS 21. 0 was used for analysis.</p><p><b>RESULTS</b>All postoperative incisions healed in one stage, the median follow-up was 34 months (range: 18-63 months). The overall local control rate was 79.7%. For patients who received the dose of more than 14 Gy versus less than 14 Gy, the local control rate was 81.6%, 75.2%, respectively (P > 0.05). For male and female, the 3 year local recurrence rate were 45.3%, 9.9% respectively (P = 0.008). Multivariate analysis showed that the sex (male versus female) was an independent prognostic factor (P = 0.036).</p><p><b>CONCLUSION</b>Surgery combined with electronic beam irradiation is a rather effective way to treat keloids. The local control rate would have a better trend if the total dose was higher than 14 Gy. Sex is an independent prognostic factor.</p>

Screening of biological amniotic immunization safety / 国际药学研究杂志

Objective To evaluate the immunity efficacy of human amniotic membranes on rats. Methods One hundred and fifty Wistar rats were randomly divided into five groups:biological amnion group,immunosuppression group,immunostimulation group, sham-operated group and blank control group. According to the study period,each group of thirty rats would be randomly divided into five experimental operation subgroups:the 1st week,the 2nd week,the 4th week,the 8th week group and the 12th week groups. The rats were implanted subcutaneously,then intramuscular injection of gentamicin sulfate for 3 days to resist the infection ,and the immune organ coefficient,and the killing abilities of NK cell ,IL-1β,IL-6 and TNF-αserum levels were detected according to the study period.Results At 1st ,2nd ,4th ,8th and 12th week after amniotic membrane implantation in rats,compared with the sham-operated and blank control groups,the biological amnion group had nonsignificant differences (P>0.05). At 1st week after amniotic membrane implantation in rats, immunosuppression group showed different levels of the immunosuppressive effect,such as the analysis of immune organ coefficient , which had significant differences compared with other groups (P<0.01). At 1st week after amniotic membrane implantation in rats,the imunostimulation group showed a certain degree of the immunostimulant effect,such as the killing abilities of NK cell,which had marked differences compared with other groups (P<0.05). Conclusion The amniotic membranes have satisfactory immune safety with implantation in rats and do not cause significant adverse immune reactions.

Pathogen distribution and risk factors of nosocomial infections in neonates in the neonatal intensive care unit / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the pathogen distribution and risk factors of nosocomial infections in neonates in the neonatal intensive care units (NICU).</p><p><b>METHODS</b>The clinical data of 145 neonates with nosocomial infection in the NICU were retrospectively reviewed.</p><p><b>RESULTS</b>Of the 145 neonates, 41 (28.3%) were infected with Klebsiella pneumoniae, 39 (26.9%) with Escherichia coli, 10 (6.9%) with Staphylococcus epidermidis, and 55 (37.9%) with other pathogens. Logistic regression analysis showed that a gestational age of ≤32 weeks (OR=5.57), birth weigh of <1500 g (OR=6.95), hospitalization time (OR=1.23), mechanical ventilation (OR=14.12) and parenteral nutrition (OR=3.01) were major risk factors for nosocomial infections caused by Klebsiella pneumoniae. The five factors were also main risk factors for nosocomial infection caused by Escherichia coli, with the OR of 3.42, 6.73, 9.96, 0.55 and 2.13 respectively. Klebsiella pneumoniae and Escherichia coli were highly resistant to β-lactam antibiotics but were relatively sensitive to levofloxacin and meropenem.</p><p><b>CONCLUSIONS</b>Klebsiella pneumoniae, Escherichia coli and Staphylococcus epidermidis are major pathogens of nosocomial infections in neonates in the NICU and they are resistant to β-lactam antibiotics. Mechanical ventilation and hospitalization time are the most important risk factors for nosocomial infections caused by Klebsiella pneumoniae and Escherichia coli respectively.</p>

Value of protein array in the diagnosis of Helicobactor pylori infection in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the value of multiple Helicobacter pylori (H.pylori) antibody detection by protein array in the diagnosis of H.pylori infection in children.</p><p><b>METHODS</b>Biopsy specimens obtained by gastroscopy from 120 children with digestive system symptoms were detected by rapid urease test (RUT) and modified Giemsa staining. Positivity in both RUT and Giemsa staining was the "gold criterion" of H.pylori infection. Serum samples of these patients were obtained and the antibodies against cytotoxin associated gene A protein (CagA), vacuolating toxin A (VacA), urease, heat shock protein 60 (Hsp60) and RdxA (nitroreductase) were detected by protein array technique.</p><p><b>RESULTS</b>H.pylori infection was identified according to the "gold criterion" in 60 children. Compared with the "gold criterion", the goodness of fit and the coefficient of contingency in the diagnosis of H.pylori infection of the following four groups antibody detection were all statistically significant (P<0.001): anti-Ure antibody alone, anti-Ure antibody combined with anti-CagA antibody, anti-Ure antibody combined with anti-VacA antibody and anti-Ure antibody combined with anti-CagA and anti-VacA antibody. The sensitivity, specificity and accuracy of the detection of anti-Ure antibody combined with anti-CagA antibody for the diagnosis of H.pylori infection were 81.7%, 91.7% and 86.7%, respectively. The antibody detection showed a high positive predictive value (90.7%) and a high negative predictive value (83.3%).</p><p><b>CONCLUSIONS</b>The antibody detection by protein array, especially the detection of anti-Ure antibody combined with anti-CagA antibody, is valuable in the diagnosis of H.pylori infection.</p>

Effect of zinc deficiency on intestinal mucosal morphology and digestive enzyme activity in growing rat / 中华儿科杂志

<p><b>OBJECTIVE</b>In this study, a growing rat model of zinc deficiency was established to investigate the effect of zinc deficiency on intestinal mucosal morphology and digestive enzyme activity as well as to provide a scientific basis for zinc supplementation therapy in patients with diarrhea.</p><p><b>METHOD</b>Three-week-old weaned Sprague-Dawley male rats (n = 30) were randomly divided into 3 groups with 10 in each: rats in the control group (ZA) were fed with a normal diet containing 30 µg/g zinc; rats in the zinc deficient group (ZD) were fed with a zinc-deficient diet containing 0.4 µg/g zinc (refer to AIN-76 formula); and rats in the paired fed group (PF) were fed with a normal diet, but the food intake was limited to intake of rats in ZD group in the previous day. All rats were provided with deionized water for drinking. Their body weight was measured and the food intake during the previous day was recorded early in the morning of the following day. Symptoms of zinc deficiency, such as anorexia, diarrhea, dermatitis, and growth retardation, were observed. Two weeks later, the rats were sacrificed and serum zinc concentration was measured. Jejunal mucosa was taken for biopsy and was stained with hematoxylin and eosin (HE). The height ratio of the jejunal mucosal villi and crypts was measured. In addition, the activity of lactase in the jejunal mucosal brush border, γ-glutamyl peptidase (GGT), and aminopeptidase N (APN) were measured.</p><p><b>RESULT</b>The average weight of the rats in the ZA, ZD, and PF groups at the beginning of the experiment was (67.4 ± 5.3) g, (64.7 ± 4.8) g, and (66.5 ± 4.1) g, respectively, and the average daily food intake was (11.2 ± 1.0) g, (11.6 ± 1.6) g, and (11.2 ± 1.4) g, respectively. The intergroup differences were not significant. On the 7(th) day of experiment, no significant differences in average food intake were observed between the ZD group and the ZA and PF groups, but the average body weight in the ZD group was significantly lower than that in the ZA and PF groups (P < 0.01). At the end of the experiment (2 weeks), the average weight in the ZD group (112.0 ± 11.5) g was significantly lower than that in the ZA (164.0 ± 15.9) g and PF groups (137.5 ± 16.2) g. The average food intake in the ZD group (13.4 ± 5.1) g was significantly lower than that in the ZA group (18.2 ± 2.4) g (P < 0.01). Serum zinc level in the ZD group (733 ± 231) µg/L was significantly lower than that in the ZA (1553 ± 159) µg/L and PF groups (1457 ± 216) µg/L (P < 0.01). The height ratio of jejunal mucosa villus and crypt in the ZA, ZD, and PF groups was 2.98 ± 0.5, 2.77 ± 0.5, and 2.81 ± 0.7, respectively, and lactase activity was (26.1 ± 15.0) U/mg, (27.4 ± 12.8) U/mg, and (40.8 ± 18.5) U/mg, respectively, without significant intergroup differences. The GGT activity in the jejunal mucosa in the ZD group (12.7 ± 6.5) U/g was significantly lower than that in the ZA (19.1 ± 10.4) U/g and PF groups (18.5 ± 7.7) U/g, but the difference was not significant. The activity of APN in the jejunal mucosa in the ZD group (25.5 ± 7.5) U/g was significantly lower than that in the ZA (48.7 ± 16.8) U/g and PF groups (43.9 ± 14.5) U/g (P < 0.01).</p><p><b>CONCLUSION</b>Zinc deficiency can cause loss of appetite, weight loss, and decreased activity of peptidase in the jejunal mucosal brush border. Zinc deficiency has little effect on the height ratio of the villus and crypt and lactase activity, thereby indicating that zinc deficiency may first affect protein digestion and absorption.</p>

Multi-slice helical CT scanning in differential diagnosis of renal clear cell carcinoma and renal papillary carcinoma / 第二军医大学学报

Objective: To evaluate the value of multi slice computed tomography (CT) in differential diagnosis of renal clear cell carcinoma and renal papillary carcinoma. Methods: The CT images of 47patients with renal cell carcinoma (RCC) were reviewed. The RCC patients were divided into 2 groups pathologically, including 37 cases of clear cell RCC and 10 cases of papillary RCC. Plain scan and three phase (corticomedullary, nephrographic and excretory phases) CT were performed in all patients. Age and sex of patients, tumor size, enhancement degree and pattern (homogeneous, heterogeneous and predominantly peripheral), the presence of calcification or cystic degeneration (necrotic or hemorrhagic areas within the tumor) and tumor spreading (including perinephric change, venous invasion and lymphadenopathy) were compared between the 2 subtypes. Results: The degrees of enhancement were significantly different between the 2 subtypes in the corticomedullary, parenchymal and excretory phases (P<0.05). Necrosis and cystic degeneration were more evident in the clear cell RCC than in papillary RCC regardless of tumor size (P<0.05). A hypervascular pattern (higher tumor enhancement after contrast material injection due to higher vascularity) was noted in 21.6% of clear cell RCC cases and in 10% of papillary RCC (P<0.05). Half of the clear cell RCC and 2.7% of papillary RCC patients showed homogeneous enhancement (P<0.05). Calcification was evident in 21.6% of clear cell RCC patients and 20% of papillary RCC patients. Conclusion: The degree of enhancement is the most valuable parameter for differentiation of clear cell RCC and papillary RCC. The presence of cystic degeneration, hemorrhage, vascularity and enhancement patterns can also contribute to the differentiation of the 2 subtypes.

Effect of lethal Vibrio vulnificus infection on blood system and pathology changes of major organs in mice / 第二军医大学学报

Objective: To investigate the effect of lethal Vibrio vulnificus infection on the blood system and the pathology changes of the major organs in mice, and to explore the possible mechanism of the related death. Methods: Lethal Vibrio vulnificus-infection model was established with mice. The model mice were divided into two groups: a control group and an infection group. ELISA was used to examine the serum levels of TNF-α, IL-1β, and TF. Serum total bilirubin (TBIL), creatinine (Cre), and blood urea nitrogen (BUN) were analyzed using automatic biochemical analyzer; whole blood cell analysis was also performed. The pathological changes of the heart, lung, liver, spleen, and kidney were observed under electron and light microscopes. Results: Compared with the control group, the serum levels of TNF-α, IL-1β, and TF were significantly increased in mice after infection with Vibrio vulnificus (P < 0.05); the serum levels of BUN, Cre, TBIL, diastase and alanine aminotransferase (ALT) were significantly increased (P < 0.05). The ratios of WBC, platelet, and lymphocytes were all significantly decreased after infection compared with the control group (P<0.05). The ratios of red blood cells, monocytes, and Hb level were significantly increased compared with the control group (P<0.05). The pathological changes of major organs included hyperaemia, edema, inflammatory cell infiltration, and apoptosis. Conclusion: Lethal infection with Vibrio vulnificus can initiate super-inflammation reaction in mice; it can also activate the blood coagulation system and induce systemic tissue injury, finally leading to death.

Recombinant proteins secreted from tissue-engineered bioartificial muscle improve cardiac dysfunction and suppress cardiomyocyte apoptosis in rats with heart failure / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Tissue-engineered bioartificial muscle-based gene therapy represents a promising approach for the treatment of heart diseases. Experimental and clinical studies suggest that systemic administration of insulin-like growth factor-1 (IGF-1) protein or overexpression of IGF-1 in the heart exerts a favorable effect on cardiovascular function. This study aimed to investigate a chronic stage after myocardial infarction (MI) and the potential therapeutic effects of delivering a human IGF-1 gene by tissue-engineered bioartificial muscles (BAMs) following coronary artery ligation in Sprague-Dawley rats.</p><p><b>METHODS</b>Ligation of the left coronary artery or sham operation was performed. Primary skeletal myoblasts were retrovirally transduced to synthesize and secrete recombinant human insulin-like growth factor-1 (rhIGF-1), and green fluorescent protein (GFP), and tissue-engineered into implantable BAMs. The rats that underwent ligation were randomly assigned to 2 groups: MI-IGF group (n = 6) and MI-GFP group (n = 6). The MI-IGF group received rhIGF-secreting BAM (IGF-BAMs) transplantation, and the MI-GFP group received GFP-secreting BAM (GFP-BAMs) transplantation. Another group of rats served as the sham operation group, which was also randomly assigned to 2 subgroups: S-IGF group (n = 6) and S-GFP group (n = 6). The S-IGF group underwent IGF-1-BAM transplantation, and S-GFP group underwent GFP-BAM transplantation. IGF-1-BAMs and GFP-BAMs were implanted subcutaneously into syngeneic rats after two weeks of operation was performed. Four weeks after the treatment, hemodynamics was performed. IGF-1 was measured by radioimmunoassay, and then the rats were sacrificed and ventricular samples were subjected to immunohistochemistry. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to examine the mRNA expression of bax and Bcl-2. TNF-α and caspase 3 expression in myocardium was examined by Western blotting.</p><p><b>RESULTS</b>Primary rat myoblasts were retrovirally transduced to secrete rhIGF-1 and tissue-engineered into implantable BAMs containing parallel arrays of postmitotic myofibers. In vitro, they secreted consistent levels of hIGF (0.4 - 1.2 µg×BAM(-1)×d(-1)). When implanted into syngeneic rat, IGF-BAMs secreted and delivered rhIGF. Four weeks after therapy, the hemodynamics was improved significantly in MI rats treated with IGF-BAMs compared with those treated with GFP-BAMs. The levels of serum IGF-1 were increased significantly in both MI and sham rats treated with IGF-BAM. The mRNA expression of bax was lower and Bcl-2 expression was higher in MI-IGF group than MI-GFP group (P < 0.05). Western blotting assay showed TNF-α and caspase 3 expression was lower in MI-IGF group than MI-GFP group after therapy.</p><p><b>CONCLUSIONS</b>rhIGF-1 significantly improves left ventricular function and suppresses cardiomyocyte apoptosis in rats with chronic heart failure. Genetically modified tissue-engineered BAMs provide a method delivering recombinant protein for the treatment of heart failure.</p>

Recombinant human growth hormone secreted from tissue-engineered bioartificial muscle improves left ventricular function in rat with acute myocardial infarction / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Experimental studies and preliminary clinical studies have suggested that growth hormone (GH) treatment may improve cardiovascular parameters in chronic heart failure (CHF). Recombinant human GH (rhGH) has been delivered by a recombinant protein, by plasmid DNA, and by genetically engineered cells with different pharmacokinetic and physiological properties. The present study aimed to examine a new method for delivery of rhGH using genetically modified bioartificial muscles (BAMs), and investigate whether the rhGH delivered by this technique improves left ventricular (LV) function in rats with CHF.</p><p><b>METHODS</b>Primary skeletal myoblasts were isolated from several Sprague-Dawley (SD) rats, cultured, purified, and retrovirally transduced to synthesize and secrete human rhGH, and tissue-engineered into implantable BAMs. Ligation of the left coronary artery or sham operation was performed. The rats that underwent ligation were randomly assigned to 2 groups: CHF control group (n = 6) and CHF treatment group (n = 6). The CHF control group received non-rhGH-secreting BAM (GFP-BAMs) transplantation, and the CHF treatment group received rhGH-secreting BAM (GH-BAMs) transplantation. Another group of rats served as the sham operation group, which was also randomly assigned to 2 subgroups: sham control group (n = 6) and sham treatment group (n = 6). The sham control group underwent GFP-BAM transplantation, and the sham treatment group underwent GH-BAM transplantation. GH-BAMs and GFP-BAMs were implanted subcutaneously into syngeneic rats with ligation of the left coronary artery or sham operation was performed. Eight weeks after the treatment, echocardiography was performed. hGH, insulin-like growth factor-1 (IGF-1) and TNF-alpha levels in rat serum were measured by radioimmunoassay and ELISA, and then the rats were killed and ventricular samples were subjected to immunohistochemistry.</p><p><b>RESULTS</b>Primary rat myoblasts were retrovirally transduced to secrete rhGH and tissue-engineered into implantable BAMs containing parallel arrays of postmitotic myofibers. In vitro, they secreted 1 to 2 microg of bioactive rhGH per day. When implanted into syngeneic rat, GH-BAMs secreted and delivered rhGH. Eight weeks after therapy, LV ejection fraction (EF) and fractional shortening (FS) were significantly higher in CHF rats treated with GH-BAMs than in those treated with GFP-BAMs ((65.0 +/- 6.5)% vs (48.1 +/- 6.8)%, P < 0.05), ((41.3 +/- 7.4)% vs (26.5 +/- 7.1)%, P < 0.05). LV end-diastolic dimension (LVEDD) was significantly lower in CHF rats treated with GH-BAM than in CHF rats treated with GFP-BAM (P < 0.05). The levels of serum GH and IGF-1 were increased significantly in both CHF and sham rats treated with GH-BAM. The level of serum TNF-alpha decreased more significantly in the CHF treatment group than in the CHF control group.</p><p><b>CONCLUSIONS</b>rhGH significantly improves LV function and prevents cardiac remodeling in rats with CHF. Genetically modified tissue-engineered bioartificial muscle provides a method delivering recombinant protein for the treatment of heart failure.</p>

Effects of transplanted myoblasts transfected with human growth hormone gene on improvement of ventricular function of rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Cell transplantation for myocardial repair is limited by early cell death. Gene therapy with human growth hormone (hGH) has been shown to promote angiogenesis and attenuate apoptosis in the experimental animal. This study was conducted to explore the effects of myoblast-based hGH gene therapy on heart function restoration and angiogenesis after myocardial infarction, and to compare the differences between myoblast-based hGH gene therapy and myoblast therapy.</p><p><b>METHODS</b>Myoblasts were isolated from several SD rats, cultured, purified, and transfected with plasmid pLghGHSN and pLgGFPSN. Radioimmunoassay (RIA) was used to detect the expression of hGH in these myoblasts. SD rats underwent the ligation of the left anterior descending coronary artery so as to establish a heart ischemia model. Thirty surviving rats that underwent ligation were randomly divided into 3 equal groups 2 weeks after left coronary artery occlusion: pLghGHSN group received myoblast infected with hGH gene transplantation; pLgGFPSN group received myoblast infected with GFP gene transplantation; control group: received cultured medium only. Four weeks after the injection the surviving rat underwent evaluation of cardiac function by echocardiography. The rats were killed and ventricular samples were undergone immunohistochemistry with hematoxylin-eosin and factor VIII. Cryosection was analyzed by fluorescence microscopy to examine the expression of green fluorescent protein. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to examine the mRNA expression of vascular endothelial growth factor (VEGF), bax and Bcl-2. hGH expression in myocardium was examined by Western blot.</p><p><b>RESULTS</b>Myoblast can be successfully isolated, cultured and transfected. The expression of hGH in transfected myoblast was demonstrated with RIA. Four weeks after therapy, the cardiac function was improved significantly in pLghGHSN group and pLgGFPSN group. Fractional shortening (FS) and ejection fraction (EF) in pLghGHSN group were elevated significantly compared with pLgGFPSN group and control group after therapy (FS: 36.9+/-5.3 vs 29.5+/-3.5, 21.8+/-2.9; EF: 56.9+/-4.3 vs 47.1+/-3.6, 38.4+/-4.8, P<0.05). Left ventricular end-diastolic dimension (LVEDD) and heart infracted size in pLghGHSN group were decreased significantly compared with pLgGFPSN group and control group after therapy (LVEDD: 5.9+/-0.3 vs 6.8+/-0.2, 8.6+/-0.3; heart infracted size: (34.5+/-4.2)% vs (40.0+/-3.9)%, (46.1+/-3.8)%, P<0.05); Green fluorescence was detected in cryosection of pLgGFPSN group. The capillary density of the pLgGFPSN group was significantly greater than those of the pLghGHSN group and control group (P<0.05). The mRNA expression of VEGF and Bcl-2/bax in pLghGHSN group was higher than in pLgGFPSN group or control group (P<0.05). The expression of hGH gene in myocardium tissue can be detected by Western blot assay in pLghGHSN group.</p><p><b>CONCLUSIONS</b>Transplantation of heart cells transfected with hGH induced greater angiogenesis and effect of antiapoptosis than transplantation of cells transfected with GFP. Combined GH gene transfer and cell transplantation provided an effective strategy for improving postinfarction ventricular function.</p>

Astrocytes protect MN9D neuronal cells against rotenone-induced oxidative stress by a glutathione-dependent mechanism / 生理学报

Astrocytes maintain homeostasis of neuronal microenvironment, provide metabolic and trophic support to neurons and modulate neuronal responses to injury. Rotenone specifically inhibits mitochondrial complex I, and long exposure to rotenone may increase the risk for Parkinson's disease (PD) and cause Parkinsonism. However, little is known about the role of astrocytes in the process of rotenone-induced dopaminergic neuron injury. In order to investigate this issue, we used MN9D cells as a cell model of dopaminergic neurons and rotenone as a toxin to initiate mitochondrial deficiency. MN9D cells treated with the normal medium or astrocyte-conditioned medium (ACM) were exposed to different concentrations of rotenone for different time followed by cell viability measurement by MTT assay. Besides, various concentrations of ACM and temporally different treatments were devised to evaluate protective efficiency of ACM. Growth curve of cells in the normal medium or ACM was continuously assessed by cell counting for 8 d. The influence of rotenone and ACM on cellular oxidative stress was determined by DCFH-DA staining followed by flow cytometric analysis. Glutathione (GSH) content after treatment of ACM or rotenone was measured by GSH assay kit. Our results showed that rotenone decreased viability of MN9D cells in a dose-dependent manner and ACM treatment significantly attenuated rotenone toxicity at each concentration. No significant difference in growth rate was observed between the normal medium and ACM treatment. Four concentrations of ACM, namely 1/3ACM, 1/2ACM, 2/3ACM and pure ACM, all displayed protection, increasing cell viability to (124.15+/-0.79)%, (126.59+/-0.82) %, (125.84+/-0.61) % and (117.15+/-1.63) % of the cells exposed directly to rotenone, respectively. Treatment with ACM through the whole experiment except the initial 24 h, 24 h before or at the same time of rotenone addition all exerted protective effects, with cell viability being (110.11+/-2.52)%, (113.30+/-2.36) %, (114.42+/-2.00)% of the cells exposed directly to rotenone, respectively. Conversely, ACM treatment 12 h after rotenone addition had no protective effect, with cell viability being (102.54+/-1.36)% of the cells exposed directly to rotenone. Moreover, ACM treatment up-regulated GSH level in MN9D cells nearly twofold. Incubation with 100 nmol/L rotenone for 24 h depleted GSH level by nearly two thirds of the control, but ACM treatment mitigated the drop of GSH level, maintaining its content at (147.83+/-0.63)% of the control. Consistent with GSH change, rotenone administration resulted in a positive rate of 96.24% of DCF staining, implying a great extent of oxidative stress, whereas treatment with ACM reduced the extent of oxidative stress to a positive rate of 78.31%. Taken together, these findings suggest that astrocytes protect MN9D cells from oxidative stress caused by rotenone, and GSH partially accounts for the protection. Therefore, astrocytes may play a protective role in the process of PD.

Multiphasic spiral CT scanning features in 100 patients with small renal cell carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the role of multiphasic spiral computed tomography (SCT) in the differential diagnosis of small renal cell carcinoma.</p><p><b>METHODS</b>The data of 100 patients with small renal cell carcinoma (< or = 3.0 cm) proved by pathology were retrospectively reviewed in order to analyze the features of SCT during plain, corticomedullary and excretory phases. There were 83 males and 17 females, with a mean age of 54. 3 years ranging from 9 to 81 years.</p><p><b>RESULTS</b>There were 38 tumor masses in the left kidney and 62 in the right one. They were 1.0-3.0 cm (mean, 2.5 cm) in the greatest dimension. According to the 2004 WHO histological classification criteria for the tumors of the kidney. Seventy-six patients had clear cell renal cell carcinoma, 4 multilocular clear cell renal cell carcinomas, 9 papillary renal cell carcinoma, 4 chromophobe renal cell carcinomas and 7 unclassified renal cell carcinomas. Clear cell renal cell carcinoma exhibited rich blood supply and inhomogeneous density due to hemorrhage, necrosis or cystic degeneration. Multilocular clear cell renal cell carcinoma presented as a multilocular cystic mass with thin wall and septa, instead of an expansile nodule. Papillary renal cell carcinoma showed inhomogeneous density and hypovascular distribution. Chromophobe renal cell carcinoma was relatively homogeneous and hypovascular. Compared with clear cell renal cell carcinoma, unclassified renal cell carcinoma showed inhomogeneous density and hypervascular distribution with more invading growth features than the other subtypes.</p><p><b>CONCLUSION</b>Commonly encountered subtypes of the small renal cell carcinoma exhibit their own specific features in multiphasic spiral CT, which may be helpful in differential diagnosis, but each subtype should be differentiated from the renal oncocytoma, cystic nephroma, complex renal cyst, renal angiomyolipoma with minimal fat and renal infiltrating urothelial carcinoma.</p>

Effects of low molecular heparin on cytokines in invasive pulmonary aspergillosis mice / 上海第二医科大学学报

Objective To investigate the effects of low molecular heparin(LWMH)on cytokines(TNF-?,IL-1? and IL-10)in blood plasma and bronchoalveolar lavage fluid of invasive pulmonary aspergillosis(IPA)mice.Methods The neutropenic IPA mouse models were established by administration of cyclophosphamide for immunologic function inhibition and intranasally challenge with Aspergillus fumigatus conidia(1?106 conidia/mouse).One hundred and twenty mice were randomly divided into 4 groups:normal control,IPA model,normal saline+LWMH and IPA+LWMH group.Normal saline+LWMH group and IPA+LWMH group received LWMH(subcutaneous injection,1 000 IU/kg,qd?2 d).Normal control and IPA model group received normal saline instedad of LWMH.At 4,8,12,24 and 48 h after inoculation,six mice were randomly taken from each group to be sacrificed.ELISA method was used to determine the concentrations of TNF-?,IL-1? and IL-10 in blood plasma and BALF.Results TNF-?,IL-1? and IL-10 in blood plasma and BALF increased significantly several hours after inoculation of conidia in IPA model and IPA+LWMH group.There were significant higher concentrations of TNF-? and IL-1? in blood plasma and BALF in IPA+LWMH group than in IPA model group(P

Effects of Panax notoginseng saponins(三七总皂苷) on extra-vascular lung water and respiratory dynamics in dog with oleic acid induced acute lung injury / 中国中西医结合急救杂志

Objective To investigate the effects of Panax notoginseng saponins(PNS,三七总皂苷) on extra-vascular lung water(EVLW) and respiratory dynamics in dog with oleic acid induced acute lung injury(ALI).Methods Eighteen Beagle dogs,intubated and mechanically ventilated with intermittent positive pressure ventilation(IPPV) mode(tidal volume(VT) 10 ml/kg, positive end-expiratory pressure(PEEP) 0,inspiratory oxygen concentration(FiO2) 1.00),were randomly assigned into three groups(each n=6): normal control group,ALI model group(induced by intravenous injection of oleic acid) and PNS group(received PNS after the ALI model was constructed).PNS 10 mg/kg being dissolved in 100 ml 5% glucose solution(GS) was pumped into central vein (2.5 ml/min) after ALI model was formed in the PNS group.Similar amount of glucose solution was given to the normal control and model groups.Respiratory dynamics and arterial blood gas(ABG) were monitored every hour.Four hours after the establishment of ALI,the dogs were sacrificed and extra-vascular lung water index(EVLWI) was quantified by a gravimetric measurement.Results In ALI dogs,PNS significantly decreased the index of EVLWI((14.10?1.45) ml/kg vs.(17.97?0.85) ml/kg,P0.05).Conclusion PNS has certain protective effect on dog with oleic acid induced ALI,it may lower EVLW and elevate the Cst total,that is beneficial to the improvement of hypoxemia.

Effects of recombinant retroviral vector mediated human insulin like growth factor-1 gene transfection on skeletal muscle growth in rat / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>This study transferred a recombinant gene encoding human insulin like growth factor-1 (hIGF-1) into modified primary skeletal myoblasts with a retroviral vector (pLgXSN) and determined whether the hIGF-1 promoted growth of skeletal muscle in rat.</p><p><b>METHODS</b>hIGF-1cDNA was amplified in vitro from normal human liver cells by using RT-PCR and cloned into plasmid vector pLgXSN. The recombinant vector pLghIGF-1SN and control vector pLgGFPSN were transfected into packaging cell PT67 and G418 was used to select positive colony. Myoblasts were infected with a high titre viral supernatant and transduction efficiency was evaluated as GFP expression. The expression of hIGF-1 mRNA in myoblasts was investigated by immunocytochemistry and RT-PCR. MTT assays detected the growth of myoblasts in vitro. Myoblasts transduced with pLghIGF-1SN were injected into hind limb muscles of 10 - 12 week male SD rats. Formed tissues were harvested 4 weeks later. Myocyte diameter, mean weight of hind limb and body were measured to evaluate the skeletal muscle growth.</p><p><b>RESULTS</b>Recombinant retroviral plasmid vector pLghIGF-1SN was constructed successfully. The titre of the packaged recombinant retrovirus was 1 x 10(6) cfu/ml. The transfection rate of PT67 cells reached 100% after G418 screening. hIGF-1 expression was positive in myoblast-IGF-1. The proliferation rate of myoblast-IGF-1 in vitro was higher than GFP-myoblast or myoblast (P < 0.05). The mean weights of hind limb and body of rats injected myoblast-IGF-1 were higher than those of the rats injected with myoblast-GFP or myoblast (P < 0.05). Myocyte diameter had a significant increase in IGF-1 group compared to GFP group and myoblast group (P < 0.05).</p><p><b>CONCLUSIONS</b>The transfection of the human IGF-1 gene mediated by a retroviral vector can promote the growth of skeletal muscle in rats. Genetically modified primary skeletal myoblasts provide a possibly effective approach to treat some skeletal muscle diseases.</p>

Structure and degradation property of the PVA-collagen complex drug membrane / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the structure and degradation property of the polyvinyl alcohol (PVA)-collagen complex drug membrane.</p><p><b>METHODS</b>Drug collagen membrane was complexed with PVA. The physical and chemical properties of the membrane were characterized by transmission electron microscopy, scanning electron microscope, forier transform-infrared spectroscopy and differential scanning calorimetry. Degradation experiment was performed to determine the degradation property of membrane and a degradation curve was therefor drawn.</p><p><b>RESULTS</b>The thermodynamic stability of collagen membrane was not destroyed by adding PVA. Collagen had good compatibility with PVA. Compared with collagen membrane, collagen-PVA complex membrane had smaller and evener pores. Adding PVA decreased the degradation rate of membrane.</p><p><b>CONCLUSIONS</b>PVA-collagen membrane has better microstructure and antidegradation property than collagen membrane.</p>