Protective Effect of Xiayuxue Tang on Renal Fibrosis Rats Based on Wnt/β-catenin and TGF-β1/Smad Signal Pathway / 中国实验方剂学杂志

Objective:To study the protective effect of Xiayuxue Tang on adenine-induced renal fibrosis model in rats and its impact on Wnt/<italic>β</italic>-catenin and transforming growth factor <italic>β</italic>₁（TGF-<italic>β</italic>₁）/Smad signal pathway. Method:A total of 50 SPF-grade male SD rats were randomly divided into 5 groups： the normal group， the model group， the losartan group （9 mg·kg^-1） and low and high dose （2.43，4.86 g·kg^-1） of Xiayuxue Tang groups. The rat model of renal fibrosis was established by ig administration adenine （250 mg·kg^-1） for 24 consecutive days. The rats were then given the corresponding drugs for 30 consecutive days. The levels of serum creatinine（SCr） and blood urea nitrogen （BUN） were measured by enzyme-linked immunosorbent assay（ELISA）. The histopathological changes of renal tissues in rats were observed by hematoxylin and eosin （HE） staining. The collagen deposition in rat renal tissue was observed by Masson staining； the protein expression levels of Wnt5a， Wnt5b， <italic>β</italic>-catenin， TGF-<italic>β</italic>₁， Smad4， Smad7 in renal tissue were detected respectively by immunohistochemistry（IHC） and Western blot. Result:Compared with the normal group， the results of each experimental group showed that SCr and BUN levels significantly increased in the model group （<italic>P</italic><0.01）. SCr and BUN levels decreased significantly after the intervention with the Xiayuxue Tang （<italic>P</italic><0.01）. Compared with the normal group， HE and Masson staining results showed that rats in the model group had severe renal interstitial damage and massive deposition of renal interstitial collagen. The renal interstitial tubule injury was relieved after the intervention with the Xiayuxue Tang， and the renal interstitial collagen deposition decreased. The results of IHC and Western blot showed that compared with the normal group， the expressions of Wnt5a， <italic>β</italic>-catenin， TGF-<italic>β</italic>₁ protein in the kidney of rats up-regulated （<italic>P</italic><0.01）， while the expressions of Wnt5b and Smad7 protein down-regulated （<italic>P</italic><0.01）. After the intervention with Xiayuxue Tang， the expressions of Wnt5a， <italic>β</italic>-catenin， TGF-<italic>β</italic>₁ protein down-regulated （<italic>P</italic><0.01）， while the expressions of Wnt5b and Smad7 protein up-regulated（<italic>P</italic><0.01）. There was no significant difference between the low-dose and high-dose groups with Xiayuxue Tang. Conclusion:Xiayuxue Tang has the protective effect on RIF rats induced by adenine， and its mechanism is related to the inhibition of Wnt/<italic>β</italic>-catenin and TGF-<italic>β</italic>₁/Smad signal pathway.

Shengyutang Improves Learning and Memory Ability of Sleep Deprived Mice by Regulating Levels of Monoamine Neurotransmitters in Hippocampus / 中国实验方剂学杂志

Objective:To observe the effect of Shengyutang on the levels of monoamine neurotransmitters in the hippocampus， and explore its possible mechanism on improving the learning and memory abilities of sleep deprivation （SD） mice. Method:The 50 mice were divided into normal group， model group， estazolam group， Shengyutang low and high dose groups， with 10 mice in each group. A multi-platform water environment was used to prepare SD mouse models. The low and high-dose Shengyutang groups received intragastric administration of 12.5， 25 g·kg^-1， respectively. The mice in the model group were intragastrically administered with the same dose of normal saline daily for 8 weeks. Morris water maze experiment was used to observe the behavioral changes of SD mice in the evasion latency period， the number of crossing platforms， and the stay time in the target quadrant of each group. HE staining was used to observe the pathomorphological changes of the hippocampal tissue of each group. The expression levels of eight monoamine neurotransmitters including serotonin （5-HT），dopandne （DA），epinephrine （EP），norepinephrine （NE），5-hydroxyindole acetic acid（5-HIAA）， high vanillic acid （HVA）， levodopa（<italic>L</italic>-DOPA），and 3，4-dihydroxyphenylacetic acid（DOPAC）were detected by high performance liquid chromatography， and the expression levels of c-Fos protein in hippocampus were detected by immunohistochemistry. Result:Compared with the normal group， the SD mice in the model group were in a poorer general state and severe fatigue was observed. Compared with the model group， SD mice in each dose group of Shengyutang got improved in eating， activity， sleep， hair color， and response to external stimuli. Compared with the normal group， the body weight of SD mice in the model group was significantly reduced （<italic>P</italic><0.05）， but the body weight in the Shengyutang high-dose group increased the most as compared with the model group （<italic>P</italic><0.05）. Compared with the normal group， the hippocampal cells in the model group were disorderly arranged， incomplete in shape， increased in gap and decreased in number. Compared with the model group， the number of neurons in the hippocampus of SD mice in each dose group of Shengyutang increased. Compared with the normal group， the escape latency time of SD mice in the model group was significantly prolonged， the times of crossing platform and the residence time in the target quadrant significantly decreased （<italic>P</italic><0.01）. Compared with the model group， the times of crossing platform and the residence time in the target quadrant of mice in each dose group of Shengyutang significantly increased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the normal group， the levels of 5-HT， 5-HIAA， <italic>L</italic>-DOPA， DOPAC， EP， NE， HVA and DA in the model group significantly decreased （<italic>P</italic><0.05，<italic> P</italic><0.01）； but these levels in each dose group of Shengyutang were higher than those in model group （<italic>P</italic><0.05）. Compared with the normal group， the average MD value of c-Fos protein in the hippocampus of the model group significantly increased （<italic>P</italic><0.01）， and the expression levels of c-Fos protein in the hippocampus of Shengyutang groups were significantly lower than those in model group （<italic>P</italic><0.01）. Conclusion:Shengyutang can improve the learning and memory abilities of SD rats， and its mechanism may be related to the decrease of monoamine neurotransmitter and c-Fos protein expression.

Optimization of Formulation of Fufang Huangqi Cream by D-optimal Mixture Design Based on AHP-CRITIC Analysis / 中国实验方剂学杂志

Objective::To optimize the matrix prescription of Fufang Huangqi cream and evaluate its rheological properties. Method::With appearance, spreadability and stability as evaluation indexes, the weighting coefficient of each evaluation index was determined by analytic hierarchy process (AHP), criteria importance through intercriteria correlation method (CRITIC) and AHP-CRITIC mixed weighting method. The formulation of Fufang Huangqi cream was optimized by D-optimal mixture design and its rheological properties were evaluated. Result::The weight coefficients of appearance, spreadability and stability according to AHP-CRITIC mixed weighting method were 0.185, 0.282 and 0.532, respectively. According to D-optimal mixture design based on AHP-CRITIC analysis, the optimized formulation of Fufang Huangqi cream was liquid paraffin of 3.70 g, vaseline of 2.00 g, stearic acid of 2.00 g, sodium dodecyl sulfate of 5.90 g, glycerin of 6.00 g and extract of 20.40 g. The rheological parameters of Fufang Huangqi cream was non-newtonian index<1, storage modulus>loss modulus. Conclusion::The preferred matrix formulation is stable and feasible. Fufang Huangqi cream has good appearance and is a shear thinning non-newtonian fluid. Its viscosity and ductility meet the needs of industrial production and clinical application.

Study on preparation technology and physical fingerprint of Carthami Flos Granules based on quality by design / 中草药

Objective: To study the prescription and preparation technology of Carthami Flos Granules and evaluate its physical quality based on quality by design. Methods: Carthami Flos was used as a model drug. The evaluation indicators were retention rate of hydroxysafflor yellow A (HSYA) and total flavonoid, primary formation rate, bulk density, tap density, relative homogeneity index, Hausner ratio, angle of repose, loss on drying, and hygroscopicity. The single factor experiments and the Box-Behnken design response surface method were used to investigate the particle forming process parameters. The physical fingerprint of granules was constructed by seven indexes including bulk density, tap density, relative homogeneity index, Hausner ratio, angle of repose, loss on drying, and hygroscopicity. Results: The optimum conditions were as follows: the dosage ratio of drug to adjuvant was 10.8, the amount of wetting agent (95% ethanol) was 32%, and dried at 70 ℃ for 120 min. The similarity of the physical fingerprint of 10 batches of Carthami Flos Granules was higher than 0.99. Conclusion: The prescription and preparation technology of Carthami Flos Granules and the physical quality were scientific and reasonable, which can provide ideas and methods for the research and development of Chinese materia medica.

Establishment and preliminary evaluation of multiple drug delivery system of effective components of Angelica sinensis / 药学学报

The multiple drug delivery system of components of traditional Chinese medicine is a system composed of multiple components and multiple units. According to the characteristics of each component, different drug delivery units are designed and combined to achieve the purpose of improving bioavailability and enhancing drug efficacy. In this study, supercritical extracts, phenolic acids, and polysaccharides derived from Angelica sinensis were examined as research objects, and a pellet-based vehicle was applied to construct a multiple drug delivery system for the treatment and chemoprevention of colitis and colorectal cancer. The extrusion-spheronization method was used to prepare pellets of Angelica polysaccharides which should be released in the stomach. The yield in 18-24 mesh and plane critical angle served as the index. The Box-Behnken design and the orthogonal design were used to optimize the formulation and parameters of pellets. According to a previous study, the colon specific pellets loading supercritical extracts and phenolic acid extracts were prepared by the optimized process. These two units of pellets were combined into the multiple drug delivery system of effective components of Angelica sinensis, and the quality evaluation and in vitro release study were conducted. The dynamic observation of pellets in mice was evaluated using small animal in vivo imaging system. The prescription of the Angelica polysaccharides gastric releasing pellets was:microcrystalline cellulose 6.5 g, polysaccharide 3.3 g, silica 0.2 g and 7 mL of 60% ethanol as wetting agent. The process parameters were as follows:extrusion rate at 75 r·min-1, rounding rate at 1 800 r·min-1, and rounding time for 3 min. Both in vivo and in vitro studies indicate that the prepared multiple drug delivery system of effective components of Angelica sinensis produced good release properties. The polysaccharide pellets could be rapidly released in the artificial gastric fluid and in the stomach. The colon specific pellets showed good targeting. They released little in the artificial gastric fluid within 2 hours, released less than 20% in the artificial intestinal fluid for 4 hours, and released more than 90% in artificial colon fluid for 6 hours.

Optimize preparation of compound licorice microemulsion with D-optimal design / 中国中药杂志

In order to increase the solubility of essential oil in compound licorice microemulsion and improve the efficacy of the decoction for treating chronic eczema, this experiment intends to prepare the decoction into microemulsion. The essential oil was used as the oil phase of the microemulsion and the extract was used as the water phase. Then the microemulsion area and maximum ratio of water capacity was obtained by plotting pseudo-ternary phase diagram, to determine the appropriate types of surfactant and cosurfactant, and Km value-the mass ratio between surfactant and cosurfactant. With particle size and skin retention of active ingredients as the index, microemulsion prescription was optimized by D-optimal design method, to investigate the release behavior of the optimized prescription. The results showed that the microemulsion was optimal with tween-80 as the surfactant and anhydrous ethanol as the cosurfactant. When the Km value was 1, the area of the microemulsion region was largest while when the concentration of extract was 0.5 g·mL⁻¹, it had lowest effect on the particle size distribution of microemulsion. The final optimized formulation was as follows: 9.4% tween-80, 9.4% anhydrous ethanol, 1.0% peppermint oil and 80.2% 0.5 g·mL⁻¹ extract. The microemulsion prepared under these conditions had a small viscosity, good stability and high skin retention of drug; in vitro release experiment showed that microemulsion had a sustained-release effect on glycyrrhizic acid and liquiritin, basically achieving the expected purpose of the project.

Observation of curative effect of hemorrhoids lotion on pain, edema and bleeding after anorectal surgery / 中国中药杂志

The purpose of this study was to evaluate the value of Chinese herbal fumigation in the postoperative anal disease. The authors randomly divided 348 patients into treatment group and control group with 174 cases in each group. The treatment group was given to the Chinese herbal medicine hemorrhoids lotion for fumigation based on conventional anti infective therapy, routine dressing change and relaxing bowel. The control group was given to 1 000 mL 1: 5 000 potassium permanganate solution for sitz bath, fumigation based on conventional anti infective therapy, routine dressing change and relaxing bowel. The pain score, edema score, bleeding score, granulation tissue growth score and wound healing time of two groups were compared after operation. The results showed that the postoperative 6 h pain scores were higher in the two groups, the postoperative 3,5,7 d pain scores gradually decreased, the difference was statistically significant (P < 0.05). The difference of postoperative 6 h pain scores was no significant difference between the two groups, while postoperative 3,5,7 d pain scores in the treatment group were significantly lower than those in the control group (P < 0.05). 7 days after operation, anal margin of edema score and blood in the stool score in the treatment group were lower than those in control group, meat medicine growth score was higher than that of the control group, the difference had statistical meaning (P < 0.05). The healing time of two groups was respectively (13.89 + 2.78), (18.45 + 1.65) d (P < 0.05). This study suggested that Chinese herbal fumigation and washing could reduce the pain degree of patients, the anal margin of edema, and the blood in the stool, also could promote granulation tissue growth and shorten the time of wound healing, deserve the clinical expansion.

Prevention and treatment of drug-related stroke / 浙江大学学报·医学版

Stroke classified into ischemic and hemorrhagic subtypes, is among the most devastating diseases for human being. Certain drugs could increase the blood viscosity, thereby increasing the potential risk of ischemia. Anti-platelet and anti-coagulation drugs, as the treatment of first choice, increase the risk of intracranial hemorrhage and death. Here, we are the first to propose the concept of drug-related stroke, and discuss the treatment strategy for patients who are complicated with intracranial hemorrhage or plan to receive neurosurgical operation during either anti-platelet or anti-coagulation medication. We hope to arouse the attention for drug related stroke among the clinicians and offer recommendation for clinical intervention.

Brainstem cavernous malformation and its surgical treatment / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To analyze the clinical characteristics and surgery approach for patients with brainstem cavernous malformation (BSCM).</p><p><b>METHODS</b>The clinical data of 23 BSCM patients (5 cases at midbrain, 16 cases at pons, and 2 cases at medulla) treated in the Second Affiliated Hospital Zhejiang University School of Medicine from July 2003 to June 2014 were retrospectively reviewed. The medical history, radiological findings, operation records, postoperative course and follow-up results were analyzed.</p><p><b>RESULTS</b>Suboccipital approach, retrosigmoid approach, subtentorial supracerebella approach, Poppen approach, pterional approach, Kawase approach, interhemispheric transcallosal third ventrical approach were applied for the surgery of BSCM patients. Among them, Kawase approach and interhemispheric transcallosal third ventrical approach were firstly reported in treatment of BSCM. Total resection was achieved in 22 cases. Neurological function was improved in 15 cases, unchanged in 7 cases and deteriorated in 1 case. Fifteen cases were followed up for a mean period of 3.5 years and signs of recurrence was found.</p><p><b>CONCLUSION</b>Proper selection of surgical approach is important to assure total resection of the lesions, to protect surrounding normal vital structures and to avoid post-surgical complications.</p>

Effect of high mobility group box protein 1 on the Kupffer cells of rats with severe burn and the role of receptor for advanced glycation end products in the process / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the effect of high mobility group box protein 1 (HMGB1) on the production of pro-inflammatory cytokines by Kupffer cell (KC) of rats with severe burn and the role of receptor for advanced glycation end products (RAGE) in the process.</p><p><b>METHODS</b>Model of 30% TBSA full-thickness burn was reproduced in 32 SD rats through immersing the back in 98°C water for 12 s. KC (32 samples) was isolated from rat liver 24 h after injury and inoculated in 24-well plate in the concentration of 1×10(6) cell per well. (1) Cells were divided into control group (cultured with 1 mL PBS) and HMGB1 group (stimulated with 100 ng/mL HMGB1 in the volume of 1 mL) according to the random number table, with 8 samples in each group. At post culture hour (PCH) 48, the expression of RAGE (denoted as grey value ratio) was detected with Western blotting. (2) Another portion of cells were divided into control group (cultured with 1 mL PBS), HMGB1 group (treated with 100 ng/mL HMGB1 in the volume of 1 mL), HMGB1 + anti-RAGE antibody group (treated with 100 ng/mL HMGB1 in the volume of 1 mL after being pre-incubated with 20 µg/mL anti-RAGE monoclonal antibody in the volume of 1 mL for 2 hours), HMGB1 + recombinant rat RAGE/Fc chimera (rrRAGE/Fc) group (treated with the mixture of 100 ng/mL HMGB1 in the volume of 0.5 mL and 5 µg/mL rrRAGE/Fc in the volume of 0.5 mL which were pre-incubated for 2 hours) according to the random number table, with 8 samples in each group. At PCH 48, the protein levels of TNF-α and IL-1β in supernatant were determined with enzyme-linked immunosorbent assay, while the mRNA expression of TNF-α and IL-1β (denoted as grey value ratio) were determined with Northern blotting. Data were processed with one-way analysis of variance, t test, and LSD test.</p><p><b>RESULTS</b>(1) The expression of RAGE in HMGB1 group (1.036 ± 0.101) was significantly higher than that of control group at PCH 48 (0.191 ± 0.024, t = -23.158, P = 0.000). (2) In HMGB1 group, HMGB1 + anti-RAGE antibody group, and HMGB1 + rrRAGE/Fc group, the contents of TNF-α in supernatant were respectively (10.59 ± 1.39), (9.91 ± 1.68), (11.51 ± 2.27) ng/mL; the contents of IL-1β in supernatant were respectively (2.49 ± 0.33), (2.08 ± 0.32), (2.42 ± 0.42) ng/mL; the mRNA levels of TNF-α in cells were respectively 0.311 ± 0.009, 0.301 ± 0.047, 0.326 ± 0.016; the mRNA levels of IL-1β in cells were respectively 0.237 ± 0.021, 0.244 ± 0.041, 0.245 ± 0.013. There were no statistically significant differences in the above indexes among these three groups (with P values all above 0.05). Their levels were all significantly higher than those of control group [with contents of TNF-α and IL-1β in supernatant respectively (2.69 ± 0.14), (0.43 ± 0.05) ng/mL, and mRNA levels of TNF-α and IL-1β in cells respectively 0.140 ± 0.022, 0.077 ± 0.005, P values all below 0.01].</p><p><b>CONCLUSIONS</b>HMGB1 can induce the production of pro-inflammatory cytokines TNF-α and IL-1β from the KC in rats with severe burn. However, RAGE does not play a predominant role in this process.</p>

Mental neurovascular V-Y island advancement flap in functional reconstruction of partial lower lip defect / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the functional reconstruction technique for partial lower lip defects.</p><p><b>METHODS</b>7 patients with lower lip cancer (3 cases of basal cell carcinoma, 2 cases squamous cell carcinoma and 2 cases papillary carcinoma) underwent excision. The full-thickness lower lip defects were one-third to two-third of the total lower lip length. All the defects were reconstructed with V-Y island advancement flaps based on the mental neurovascular bundle.</p><p><b>RESULTS</b>There was no flap loss. No recurrence was observed during the follow-up period of 3 months to one year. Both the aesthetic appearance, muscle function and sensation of the reconstructed lower lip were satisfactory.</p><p><b>CONCLUSIONS</b>Mental neurovascular V-Y island advancement flap is an ideal method for functional repair of partial lower lip defect.</p>

Reverse facial artery-submental artery island myocutaneous flap for reconstruction of oral and maxillofacial defects following cancer ablation / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the reliability of the reverse facial artery-submental artery island myocutaneous flap for repairing the oral and maxillofacial defects.</p><p><b>METHODS</b>Eighteen oral and maxillofacial defects were repaired with reverse facial artery-submental artery island myocutaneous flaps following resection of malignant tumors. The ages of the patients ranged from 28 to 90 years; 11 were male and 7 were female. Primary sites of the lesions were the tongue squamous cell carcinoma (SCC) (7 cases), buccal mucosa SCC (4 cases), palate SCC (3 cases), oropharyngeal SCC (2 case), and facial skin basal cell carcinoma (2 cases). The sizes of skin paddle varied from a minimum of 4.0 cm x 12.0 cm to a maximum of 5.0 cm x 15.0 cm. The followed-up period was 9 to 18 months (11.8 months on average).</p><p><b>RESULTS</b>The postoperative outcome for the flaps was 17 cases surviving, one complete necrosis, and one temporary palsy of the marginal mandibular branch of the facial nerve. The form and function of recipient sites were well recovered. The donor site left a well-hidden scar. One case of cervical lymph node metastasis was observed.</p><p><b>CONCLUSIONS</b>Reverse facial artery-submental artery island myocutaneous flap is reliable for reconstructing medium-sized oral and maxillofacial defects.</p>

Replication of Bombyx mori Densonucleosis Virus (Zhenjiang isolate) in different silkworm strains / 生物工程学报

The invertebrate parvovirus Bombyx mori Densonucleosis Virus type 3(Zhenjiang isolate), named BmDNV-3, is a kind of bidensovirus. The most obvious characteristic in the genome of BmDNV-3 is that it has 2 sets of DNA molecular (VD1, VD2),and each of them is encapsidated respectively in the form of single-stranded liner DNA ( + VD1, - VD1, + VD2, - VD2) in equal percentage. So the BmDNV-3 has 4 kinds of virions. Furthermore the sequence of BmDNV-3 is able to encode DNA polymerase itself. Some strains of silkworm revealed complete resistance to BmDNV-3, so they didn' t fall sick. To investigate the difference in the process of infection and replication between the 2 virions ( VD1, VD2) of this bidensovirus, and the difference of the increment in the resistant or susceptible host, the 5th instar larvae of the susceptible silkworm strain (HUABA 35) and the resistant silkworm strain(QIUFENG d) were inoculated determinate dose of BmDNV-3 by oral ingestion. Then the midgut were collected at 9 timepoints. The silkworm cytoplasm actin A3 was used to be normalized gene, so the number of cells in collected tissue could be determined. The result shows that whatever in the susceptible silkworm strain or in the resistant one, the copies of VD1 and VD2 in the genome of BmDNV-3 collected at the different timepoint were almost at the equal level respectively, so that the VD1 and VD2 were replicated with synchronization. The process of infection in the susceptible silkworm strain was devided into 3 partitions, latent period( 2 - 12 hours post inoculation), exponential phase (12 - 36 hours post inoculation)and stationary phase (36- 96 hours post inoculation and there are about 2 x 10(5) copies per cell) . In the resistant silkworm strain, the virus were replicated at a very low level, that was from 6 - 10 copies 2 hours post inoculation to 150 - 200 copies 96 hours post inoculation (about 20 times) . So we predict that the resistance in some of the silkworm strains from BmDNV-3 was a kind of chronic representation that the host carried virus without being caused flacherie.

Nudivirus Genomics: Diversity and Classification / 中国病毒学

Nudiviruses represent a diverse group of arthropod specific, rod-shaped and dsDNA viruses. Due to similarities in pathology and morphology to members of the family Baculoviridae, they have been previously classified as the so-called "non-occluded" baculoviruses. However, presently they are taxonomically orphaned and are not assigned to any virus family because of the lack of genetic relatedness to Baculoviridae,. Here, we report on recent progress in the genomic analysis of Heliothis zea nudivirus 1 (HzNV-1), Oryctes rhinoceros nudivirus (OrNV), Gryllus bimaculatus nudivirus (GbNV) and Heliotis zea nudivirus 2 (HzNV-2). Gene content comparison and phylogenetic analyses indicated that the viruses share 15 core genes with baculoviruses and form a monophyletic sister group to them. Consequences of the genetic relationship are discussed for the classification of nudiviruses.

Organization and transcription strategy of genome of Bombyx mori bidensovirus (China isolate) VD1 / 生物工程学报

To better understand the genomic structure and function of Bombyx mori bidensovirus (China isolate) VD1, the VD1 was purified and cloned into the pUC119 vector, and the complete nucleotide sequence of VD1 was determined. Sequence analysis showed that VD1 genome consisted of 6543 nts including inverted terminal repeats (ITRs) of 224 nts. In the viral genome, three major open reading frames (ORF1, ORF2 and ORF3) in the plus strand and one major ORF (ORF4) in the complementary strand were identified. Comparison of the complete genome sequence between Bombyx mori bidensovirus (China isolate) and BmDNV-2 (Yamanashi isolate) showed an identity of 98.4% in VD1, with a total number of 104 bp substitutions and 1 bp insertions found in Bombyx mori bidensovirus (China isolate), the highly variable regions were mainly located in VD1 ORF3 and VD1 ORF4. Northern blotting revealed that VD1 contained 1.1 kb and 1.5 kb transcript in the left-half 'plus' strand, and one transcript about 3.3 kb of 'minus' strand in the right-half. Sequencing of 3' and 5' ends of transcript products showed the 1.1 kb transcript started at nt 290 and ended at nt 1437, the 1.5 kb transcript was found to start nt 1423 and ended at 2931, and the 3.3 kb transcript was found to start nt 6287 and ended at nt 2922. Therefore, the 1.5 kb transcript in the left-half plus' strand and 3.3 kb transcripts of minus' strand in the right-half overlapped for 10 nts at the 3' ends. These results indicate that this virus employs a transcription strategy that is radically different from that of the other reported DNVs.

Combined repair of large defect caused by radical surgery of advanced tongue cancer with rib-major pectoralis myocutaneous flap carrying costal parietal pleura / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the clinical value and safety of using rib-major pectoralis myocutaneous flap carrying costal parietal pleura in combined repair of large soft and hard tissue defect caused by radical surgery of advanced tongue cancer.</p><p><b>METHODS</b>Six patients with advanced tongue carcinoma involving the floor of mouth and mandible were performed combined radical neck dissection with glossectomy and mandibulectomy, which caused large soft and hard tissue defect. Six rib-major pectoralis myocutaneous flaps carrying costal parietal pleura were transferred for immediate repair of the large defects. The rib flaps were applied for the repair of mandible, and the major pectoralis myocutaneous flaps were applied for the reconstruction of tongue and floor of mouth.</p><p><b>RESULTS</b>Six patients recovered well after operation. Six rib-major pectoralis myocutaneous flaps carrying costal parietal pleura survived well; the wounds of surgical incision of the oral cavity, neck, and chest healed up. The reconstructed tongue and the lower face appearance were satisfactory, the occlusion relationships were normal; the speaking as well as swallowing functions recovered.</p><p><b>CONCLUSIONS</b>It's safe and reliable to use rib-major pectoralis myocutaneous flap carrying costal parietal pleura to repair large soft and hard tissue defect in oral and maxillofacial region. Opening pleural cavity and harvest costal parietal pleura would not influence patients' thoracic movement and breath function and would not cause other complications. It's simple and safe for harvesting the composite flap. Carrying costal parietal pleura assures the sufficient blood supply of rib in the composite flap.</p>

Interaction between p38 mitogen-activated protein kinase signal transduction pathway and NF-kappaB/IkappaB system on the proinflammatory cytokines release after burn trauma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the interaction between p38 mitogen-activated protein kinase signal transduction pathway and nuclear factor (NF)-kappaB/IkappaB system on the proinflammatory cytokines release after burn trauma.</p><p><b>METHODS</b>Human monocyte line THP-1 were incubated with serum from eight healthy controls, burn sera, burn sera pretreatment with SB203580, and burn sera pretreatment with pyrrolidine dithiocarbamate (PDTC). After 24 hours incubation with serum, tumor necrosis factor (TNF)-alpha and interleukin-1beta (IL-1beta) levels in THP-1 culture supernatants were measured by ELISA. The activities of p38 MAPK and expressions of IkappaBalpha in THP-1 were measured by Western blot analysis. The EMSA method was used to characterize the binding activities of NF-kappaB and activating protein (AP)-1 in THP-1.</p><p><b>RESULTS</b>In comparison with normal controls, burn sera resulted in a significant higher level release of TNF-alpha and IL-1beta in THP-1 [(7.30 +/- 0.84) ng/ml vs (2.20 +/- 0.28) ng/ml, P < 0.05; (2.88 +/- 0.38) ng/ml vs (0.81 +/- 0.14) ng/ml, P < 0.05], which were significantly inhibited by pretreatment with SB203580 or PDTC. Burn sera showed increased activities of p38 MAPK and AP-1 in THP-1 (4728 +/- 582 vs 1291 +/- 163, P < 0.05; 946 +/- 137 vs 361 +/- 40, P < 0.05), which were abolished by pretreatment with SB203580 but not PDTC. The expression of IkappaBalpha in THP-1 incubated with burn sera was significantly decreased than those incubated with control sera (1211 +/- 115 vs 2658 +/- 318, P < 0.05), which were abolished by pretreatment with PDTC but not SB203580. Burn sera also leaded to an increased activity of NF-kappaB in THP-1 (1636 +/- 170 vs 317 +/- 32, P < 0.05), which were abolished by pretreatment with PDTC but not SB203580.</p><p><b>CONCLUSIONS</b>There are no direct interaction between p38 MAPK signal transduction pathway and NF-kappaB/IkappaB pathway. These two pathways, which regulate the production of TNF-alpha and IL-1beta in monocyte following burn trauma, are parallel and independent.</p>

Role of p38MAPK signal transduction pathway in Kupffer cells production of TNF-alpha and IL-1beta in severely burned rats / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the role of p38 mitogen-activated protein kinase (MAPK) signal transduction pathway in the Kupffer cells production of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta in severely burns rats.</p><p><b>METHODS</b>Male health adult Sprague-Dawley rats were randomized into four groups: sham burn rats given vehicle, sham burn rats given the p38 MAP kinase inhibitor SB203580, rats given a 30% total body surface area (TBSA) full-thickness burn and fluid resuscitation plus vehicle, and burn rats given injury and fluid resuscitation plus SB203580. Rats from each group were killed at 24 h after burn or sham burn and Kupffer cells (KCs) were isolated. After 18 h incubation, KCs next were stimulated with 50 ng/ml of LPS for 18 h. After stimulation, supernatants were removed for analysis of TNF-alpha and IL-1beta levels by ELISA. The TNF-alpha and IL-1beta mRNA expressions (by quantitative real-time RT-PCR) and the activities of p38 MAPK and JNK (by Western blot analysis) in KCs were examined.</p><p><b>RESULTS</b>Eighteen hours after 50 ng/ml LPS stimulation, KCs from burn rats released significantly higher levels of TNF-alpha and IL-1beta than did shams. The mRNA levels of TNF-alpha and IL-1beta in KCs increased significantly postburn. Western blot analysis suggested that expression of phosphorylated p38 MAPK and JNK were increased in KCs harvested from burn group after stimulation with LPS compared with those from sham group. In vivo administration of SB203580 markedly suppressed both the release of TNF-alpha and IL-1beta and the mRNA expressions of TNF-alpha and IL-1beta in KCs from both sham and burn rats. p38 MAPK activity in KCs was abolished by administration with SB203580, whereas JNK was not.</p><p><b>CONCLUSIONS</b>p38 MAPK signal transduction pathway mediates KCs production of proinflammatory cytokines TNF-alpha and IL-1beta in severely burned rats.</p>

The modulating role of p38 mitogen-activated protein kinase in the expression of tumor necrosis factor-alpha in hepatic cells and its role in hepatic injury in severely burned rats / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate The modulating role of p38 mitogen-activated protein kinase (MAPK) in the expression of tumor necrosis factor-alpha in hepatic cells and its role in hepatic injury in severely burned rats.</p><p><b>METHODS</b>Twenty-four adult healthy male SD rats were randomly divided into three groups (8 rats in each group): sham group, burn group, and burn with SB203580 group. A rat model of full-thickness burn injury covering 30% total body surface area (TBSA) was reproduced. The specific inhibitor of p38MAPK (SB203580 in 10 mg/kg) was given to the rats in the burn with SB203580 group at 15 minutes and 12 hours after burn. The serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured at 24 postburn hours (PBHs). The TNF-alpha mRNA expression in the liver was determined by real-time reverse transcription polymerase chain reaction, and the expression levels of p38MAPK and phosphor-p38MAPK in the liver were determined by Western blot analysis.</p><p><b>RESULTS</b>The serum levels of AST and ALT, and the expression of TNF-alpha mRNA in liver cells were significantly higher in burn group than those in sham and SB203580 groups (P < 0.05 or 0.01), but there was no difference between the two latter groups. It was indicated by Western blot results that there was no difference of p38MAPK expression in rat liver among the three groups (P > 0.05). The phospho-p38MAPK expression ratio among sham, burn and burn with SB203580 groups was 1.00:3.90:1.10. The phospho-p38MAPK expression was significantly lower in burn with SB203580 group than that in burn group (P < 0.01), but there was no significant difference compared with that in sham group (P > 0.05).</p><p><b>CONCLUSION</b>The postburn activated p38MAPK in rat liver after severe burn injury enhances the expression of TNF-alpha mRNA and participates in the development of postburn hepatic injury.</p>

Activation of p38 MAPK signal transduction pathway by burn serum and the expression of VCAM-1 in HUVECs induced by NF-kappaB / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the influence of burn serum on the expression of vascular cell adhesion molecule-1 (VCAM-1) of human umbilical vein endothelial cells (HUVECs) andits signal transduction mechanism.</p><p><b>METHODS</b>HUVECs cultured in vitro were employed for the experiment, and were divided into normal control (NC, with addition of normal serum), burn serum (BS, with addition of burn serum), SB203580 (with addition of 10 micromol/L SB203580 treatment 1 hour before burn serum treatment) and PDTC [with 10 mmol/L pyrrolidine dithiocarbamate (PDTC) 1 hour before burn serum treatment] groups. Protein and mRNA expression of VCAM-1 in HUVECs was measured by flow cytometry and reverse transcription polymerase chain reaction (RT-PCR) respectively at 0, 6, 12, 24 and 36 hours after burn serum treatment. The expression of VCAM-1 on HUVEC surface and the soluble VCAM-1 (sVCAM-1) content in HUVECs culture supernatants were measured by ELISA at 24 hours after the serum stimulation. Adherence of peripheral blood mononuclear leukocytes (PBMC) adherence to HUVECs was also observed in vitro.</p><p><b>RESULTS</b>The expression of VCAM-1 mRNA increased obviously in BS group after the burn serum stimulation and reached peak level at 24 post stimulation hour (PSH), and it decreased thereafter. The above expression was significantly decreased in SB203580 and PDTC groups at 24 PSH, but there was no difference compared with normal control (P > 0.05). The VCAM-1 expression on the membrane of HUVEC was evidently higher in BS group (66.5 +/- 6.2) than that in NC group (19.1 +/- 1.9, P < 0.05) at 24 PSH, but it was decreased significantly in SB203580 (21.7 +/- 2.3) and PDTC (23.1 +/- 2.4) groups and there was no significant difference compared with NC group (P > 0.05), and which was evidently lower than that in BS group (P < 0.05). The VCAM-1 content in the supernatant of BS group (125 +/- 10 ng/L) was obviously higher than that in NC (23 +/- 3 ng/L), SB203580 (27 +/- 5 ng/L) and PDTC (29 +/- 5 ng/L) groups. (P < 0.05). The number of PBMCs adherent to HUVECs in BS group [(197 +/- 11)%] was much larger than that in NC group [(100 +/- 4)%], SB203580 group [(113 +/- 7)%] or PDTC group [(97 +/- 112)%] at 24 PSH (P < 0.05), but no difference between NC group and SB203580, PDTC groups (P > 0.05).</p><p><b>CONCLUSION</b>Burn serum can enhance the expression of VCAM-1 in HUVECs through p38 MAPK signaling pathway, and the activation of NF-kappaB was also involved in this process.</p>