International research advances and current situation of industrialization in the last decade on the biological character and inducing conditions of natural astaxanthin accumulation in Haematococcus pluvialis cells are reviewed in present paper. Current situation of mass cultivation of Haematococcus pluvialis for industrial production of natural astaxanthin by use of photobioreactors is described. The industrialization prospect of astaxanthin in China is also proposed.

Objective To observe the effect of vein anastomosis by microvascular anastomotic device in head and neck reconstruction.Methods Sixteen cases needing free flaps were randomly selected in head and neck cancer patients with locally advanced lesion.Their veins were anastomosed by microvascular anastomotic device.Other 40 cases whose veins were anastomosed by manual anastomosis were selected as control during the same year.The statistical data of the time of veins anastomosis,thrombosis and the rate of flaps salvage were compared.Results The average time of venous anastomosis was 6.31 ± 0.87 minutes,which was lower than the manual anastomosis (19.20 ±4.62 minutes) significantly.There was no thrombosis,which was not significant different with the manual anastomosis.One case's venous blood flow was not smooth for the angled vein and the flap salvage operation was successful.Conclusion The time of venous anastomosis is shorten significantly by microvascular anastomotic device.The incidence of thrombosis is the same compared with venous anastomoses performed with sutures.The microvascular anastomotic device system allowed a time-efficient and safe venous anastomosis.

In this study, the effect of prophylactic anti-inflammation on the development of smoke-induced emphysema was investigated. Young male guinea-pigs aged 1.5-2 months (weighing 198.3+/-26.9 g) were randomly divided into 4 groups: group A (cigarette smoke exposure only), group B (cigarette smoke exposure plus pentoxifylline-rich (PTX, 10 mg/d) forage feeding), group C (cigarette smoke exposure plus intermittent cortical steroid injection (Triamcinolone acetonide, 3 mg, i.m., every three weeks) and control group (group D: animals with sham smoke exposure, raised under the same conditions). Animals in group A, B and C were exposed to smoke of cigarettes for 1 to 1.5 h twice a day, 5 days a week. All animals were killed at the 16th week and followed by morphometrical analysis of the midsagittal sectioned lung slices. Smoke exposure of 16 weeks resulted in visible emphysematous development in Group A but not in Group B and C. It was evidenced by the indicator of air-space size, mean linear intercept (Lm): 120.6+/-16.0 microm in Group A; 89.8+/-9.2 microm in Group B and 102.4+/-17.7 microm in Group C. The average Lm in either group B or group C was shorter than that in Group A (ANOVA and Newman-Keuls test, F=8.80, P=0.0002) but comparable to that (94.8+/-13.2 microm) in group D (P>0.05). It is concluded that long-term prophylactic anti-inflammation inhibits pulmonary emphysema induced by cigarette smoking in the guinea pigs.
Anti-Inflammatory Agents/*pharmacology ; Pentoxifylline/pharmacology ; Pulmonary Emphysema/etiology ; Pulmonary Emphysema/pathology ; Pulmonary Emphysema/*prevention & control ; Random Allocation ; Smoking/*adverse effects ; Triamcinolone Acetonide/*pharmacology

Objective:To investigate the effect of licochalcone A on osteoarthritis in rats and its relationship with p38-MAPK inflammatory signaling pathway.Methods:A total of 160 male Wistar rats were randomly divided into blank non-intervention, blank intervention, arthritis non- intervention and arthritis intervention groups with 40 rats in each group. Rats in the arthritis groups were subjected to unilateral anterior cruciate ligament transection, while rats in the blank groups were only subjected to skin incision and suture. Rats in the intervention groups were treated by intra-articular injection of 1 mL 10 μmol/L licochalcone A for 8 successive weeks. Eight weeks later, the cartilage of rats in each group was stained with safranin, and osteoarthritis soft tissue was scored according to Osteoarthritis Research Society International guideline under the optical microscope. The cartilage was cultured in low glucose cell culture medium supplemented with 5% fetal bovine serum for 48 hours. The contents of nitric oxide, prostaglandin E 2, sulfated glycosaminoglycan and collagen II in the medium were determined by the chemiluminescence reaction method. The expression levels of p38, phosphorylated p38 (p-p38) and matrix metalloproteinase in cartilage tissue were detected by western blot assay. Results:The progress of osteoarthritis in rats treated with licochalcone A was slow. The Osteoarthritis Research Society International score in the arthritis intervention group was significantly lower than that in the arthritis non-intervention group [(3.8 ± 1.7) points vs. (9.7 ± 1.2) points, P = 0.0064]. The contents of nitric oxide, prostaglandin E 2, sulfated glycosaminoglycan, and collagen II in the arthritis intervention group were (77.84 ± 17.65) μmol/mg and (6.78 ± 1.76) ng/mg, (89.78 ± 9.76) μg/mg, and (1.78 ± 0.76) μg/mg, respectively, which were significantly lower than those in the arthritis non-intervention group [(107.56 ± 18.74) μmol/mg, (10.756 ± 1.87) ng/mg, (125.75 ± 8.87) μg/mg, (3.76 ± 0.88) μg/mg, (NO: P = 0.002; PGE 2: P < 0.001; sGAG: P < 0.001; Collagen II: P < 0.001). Western blot assay results revealed that the relative expression of p38, p-p38, p-p38 to total p38 ratio, matrix metalloproteinase in the arthritis intervention group were (3 454 ± 421), (2 072 ± 175), (0.65 ± 0.14 )and (1 776 ± 765), respectively, which were significantly lower than those in the arthritis non-intervention group (5 322 ± 323), (4 257 ± 184), (0.89 ± 0.11), (3 865 ± 874)( p38: P < 0.001; p-p38: P < 0.001; p-p38/p38: P = 0.002; MMP: P = 0.001). Conclusion:Licochalcone A can delay the progression of osteoarthritis in rats with osteoarthritis through inhibiting inflammatory reaction and cartilage matrix degradation, and p38-MAPK signaling pathway may be involved in the regulation process.

Objective To investigate the effect of gastrointestinal Roux-en-Y gastric bypass surgery on blood sugar and insulin function of patients with type-2 diabetes mellitus.Methods Twenty-seven cases of gastric cancer patients with type-2 diabetes and undergone Roux-en-Y bypass the gastrointestinal treatment in the centre hospital of Cangzhou were selected as our subject.Body mass index (BMI),Glycosylated hemoglobin (HBA1c),Fasting and glucose (FPG),fasting insulin (FINS),Fasting C-peptide (FCP) levels were measured.Glucose (2 hPG),insulin (2 hINS) and C-peptide (2 hCP) levels were detected after 2 h for oral use 75 g glucose.Homeostasis model was applied to assess insulin resistance index (HOMA-IR).Results No significant change was seen in terms of BMI between before and after surgery.Compared to before surgery,the levels of FPG((7.58 ±0.84) mmol/L),2 hPG((10.43 ± 1.88) mmol/L),HbA1c((7.56 ± 1.15)％) and HOMA-IR(4.55 ±0.76) were lower in patients at 3 months after surgery ((9.93 ± 1.57) mtmol/L,(13.89± 2.13) mtmol/L,(9.88 ± 1.66) ％,(4.55 ± 0.76),respectively,P ＜ 0.05 or P ＜ 0.01).FPG ((6.56± 0.80) mmol/L),2 hPG ((8.57 ± 1.32) mmol/L),HbA1 c ((6.37 ± 1.24) ％),HOMA-IR (4.03 ± 0.45)of patients after 6 months were lower than that of before surgery and 3 months after surgery (P ＜ 0.05 or P＜0.01).However,the levels of FINS ((13.67 ± 1.96) mU/L),FCP((2.62 ±0.87) μg/L),2 hINS((49.91± 5.14) mU/L) and 2 hCP ((6.28 ± 1.65) μg/L) were higher in patients with 3 months after surgery compared to that of before surgery ((11.08 ± 1.69) mU/L,(1.78 ± 0.61) μg/L,(36.05 ± 4.03) mU/L,(4.28 ± 1.48) μg/L,P ＜ 0.01).Meanwhile those indices after 6 months (FINS:(15.88 ± 2.05) mU/L,FCP:(3.30 ±0.68) μg/L,2 hINS:(67.40 ±5.68) mU/L,2 hCP:(9.39 ± 1.52) μg/L) were higher than that of before surgery and 3 months after surgery(P ＜ 0.01).Conclusion Roux-en-Y gastrointestinal bypass can effectively reduce blood sugar level and improve the situation of Pancreatic Beta-cell function of gastric cancer patients with type-2 diabetes.

Objective To investigate the relationship between obstructive sleep apneahypopnea syndrome (OSAHS) and carotid stenosis in patients with ischemic cerebrovascular disease (ICVD) and to provide reference for developing the intervention strategy of carotid stenosis. Methods Eighty-seven patients with ICVD were screened from Nanjing Stroke Registry Program. The patients were divided into without (n=21), mild(n=24), moderate (n=27) and severe (n = 11) OSAHS groups according to the apnea-hypopnea index (AHI); in addition, the patients were divided into with (n =34) and without carotid stenosis (n=49) groups according to the results of digital subtraction angiography (DSA). The effects of the risk factors for cerebrovascular diseases and OSAHS on carotid stenosis in patients with ICDV were analyzed.Results There were significant differences in the proportions of alcohol consumption (χ2=8.56, P =0. 036), hypertension (χ2 = 13.20, P =0. 004) and carotid stenosis (χ2 =22.97, P =0. 006) between the no OSAHS and the mild, moderate and severe OSAHS groups. The univariate analysis showed that age (OR = 1. 066, 95% CI 1. 023- 1.112; P = 0. 003),hypertension (OR =3.587, 95% CI 1. 294- 9. 949; P =0. 014), alcohol consumption (OR =5.275,95% CI 1.855-15.001; P= 0.002) and OSAHS (OR= 1.073, 95% CI 1.033-1.115; P = 0. 000) were the risk factors for carotid stenosis. The multivariate logistic regression analysis showed that age (OR = 1. 113, 95% CI 1. 047-1. 182; P =0. 001), OSAHS (OR = 1. 096, 95% CI 1. 034-1. 160; P = 0. 000), and alcohol consumption (OR = 4. 292,95% CI 1. 217-15. 139; P = 0. 024) were the independent risk factors for carotid stenosis.Spearman rank correlation analysis suggested that the AHI levels were positively correlated with the degree of carotid stenosis (r = 0. 435, P = 0. 000). There were significant differences among the without stenosis (n =34), unilateral stenosis (n =22), and bilateral stenosis (n=27)groups (12.97 ± 10.04 vs. 21.40 ± 16.38 vs. 29.33 ± 13.81, F= 11.64, P<0.01).Conclusions OSAHS is an independent risk factor for carotid stenosis and it was positively correlated with the severity of carotid stenosis. AHI may reflect the degree of carotid stenosis and the range of neck vascular involvement to some extent.

Objective To investigate the expression of COX-2,MMP-2 and TIMP-2 ,the pathological fea-tures ,and their relationship in breast cancer. Methods The expressions of COX-2 ,MMP-2 and TIMP-2 were deter-mined by S-P immunohistochemical method on tissue chips,which containing 127 cases of breast carcinoma. Results The positive rates of COX-2,MMP-2 and TIMP-2 protein were 81.1 (103/127)% ,96.9(123/127)% and 60.6 (77/127) % respectively;The expression of COX-2 was positively related to auxiliary lymph node metastasis and TNM staging (P<0.01 and P<0.05, respectively), and inversely related to PR expression (P<0.05). Further-more,the expression of COX-2 was positively correlated with MMP-2 (r=0. 290 ,P<0.01). Conclusions The ex-pression of COX-2 might be closely related to the invasion and metastasis of breast cancer and has a close relation-ship with MMP-2. The levels of MMP-2 might be partly regulated by COX-2.

Objective To evaluate the effects of dezocine on diabetic neuropathic pain (DNP ) and expression of NMDA receptor subunit 2B (NR2B) in the spinal dorsal horns of rats .Methods Forty-eight male Sprague-Dawley rats , aged 4 weeks , weighing 150-170 g , with DNP induced by intraperitoneal injection of streptozocin (STZ) 50 mg/kg (successful induction of diabetes was defined as blood glucose >16.7 mmol/L) , were randomly divided into 2 groups ( n=24 each) using a random number table:DNP group and dezocine group (group D) .Twenty-four normal rats were chosen and served as normal control group (group C) .In group D , dezocine 2.52 mg/kg was injected intramuscularly once a day for 7 consecutive days starting from 2nd week after STZ injection ,while the rats in DNP and C groups received the equal volume of normal saline .Paw withdrawl threshold (PWT) to mechanical stimulation was measured before dezocine injection (T0 ) ,and on 1st ,3rd ,5th and 7th days after dezocine injection (T1-4 ) and on 7th day after the end of dezocine injection (T5 ) .Twelve rats in each group were sacrificed after measurement of PWT at T4 ,and T5 .The lumbar segments of the spinal cord were removed for determination of NR2B protein expression (by immuno-histochemistry and Western blot ) and NR2B mRNA expression (by RT-PCR ) in the spinal dorsal horns .Results Compared with group C ,the PWT at T0-5 in group DNP and at T0 and T5 in group D was significantly decreased , and the expression of NR2B protein and mRNA at T4 ,5 in DNP group and at T5 in D group was up-regulated ( P<0.05) .Compared with group DNP ,the PWT was significantly increased at T1-4 ,the expression of NR2B protein and mRNA was down-regulated at T4 ( P<0.05) ,and no significant changes were found in the parameters mentioned above at T5 in group D ( P>0.05) . The PWT was significantly lower at T0 and T5 ,and the expression of NR2B protein and mRNA was higher at T5 than at T4 in group D ( P<0.05 ) .Conclusion Dezocine can effectively relieve DNP in rats and inhibition of NR2B expression in the spinal dorsal horns is involved in the mechanism .

Objective To study the role of endothelial cells on the inflammatory cytokine release in septic shock through the septic shock serum stimulating human primary endothelial cells (HPAEC) and peripheral blood mononuclear cells(PBMC).Methods PBMC isolated from healthy people by density gradient centrifugation.HPAEC cell surface markers CD144 and von Willebrand factor(vWF) molecule expression by RT-PCR and Western blot.Serum levels of IL-6,TNF-α,MCP-1 from septic shock patients and healthy human detected by ELISA.HPAEC and PBMC were stimulated with the isolated serums and LPS,respectively.ELISA was used to detect the supernatant IL-6,TNF-α,MCP-1 levels.HPAEC membrane molecules ICAM-1 expression was detected by flow cytometry with serum shock and LPS stimulation.Supernatant levels of IL-6,TNF-α,MCP-1 of HPAEC with S1P1 receptor agonist CYM-5442 pretreatment was detected by ELISA after shock serum stimulation.Results Endothelial cell markers CD144 and vWF molecules could be detected in the HPAEC.Levels of inflammatory cytokines IL-6,TNF-α,MCP-1 in patients with septic shock serum were significantly higher than healthy people (P＜0.01).PBMC and HPAEC with LPS or shock serum treatment respectively,compared with normal group,levels of inflammatory cytokines in the culture supernatant were significantly higher(P＜0.01).For PBMC,the level of inflammatory cytokines between shock group and LPS group were not significantly different (P＞0.05).But for HPAEC,levels of inflammatory cytokines in the supernatant of the shock group compared to the LPS group was significantly higher (P＜0.01).Similarly,when two cells after LPS stimulation,IL-6,TNF-α levels of HPAEC's supernatant were significantly lower than PBMC' s (P＜0.01),MCP-1 levels was no difference (P＞ 0.05).But when the stimulation of shock serum,HPAEC of IL-6,TNF-α levels and PBMC no significant difference (P ＞0.05).MCP-1 was significantly increased (P＜0.01).Shock patients serum stimulation S1P1 receptorspecific agonist CYM-5442 pretreatment of HPAEC with pretreatment of S1P1 receptor specific agonist CYM-5442,the culture supernatant of inflammatory cytokines IL-6,TNF-α,MCP-1 levels were significantly lower (P＜0.01).Conclusion Endothelial cells may play a central role on the release of inflammatory cytokine during septic shock.

Objective To analyze and determine the efficient T- and B-combined (T/B) antigenic epitopes in Helicobacter pylori adhesin A. Methods Recombinant HpaA (rHpaA) was expressed for immunizing rabbit to generate antiserum. T- and B-cell epitopes in HpaA molecule were predicted by using bioinformatic technique. The segments to encode T/B combined epitope peptides were amplified by PCR and the phage display systems of T/B combined epitopes were subsequently constructed. PEG/NaCl precipitation method was applied to extract the recombinant phage PⅢ (rPⅢ) that displayed T/B combined epitopes. By using either commercial IgG against whole-cell of Helicobacter pylori or rHpaA antiserum as the primary antibody, the T/B combined epitopes displayed in rP Ⅲ s were screened and identified by Western blot and ELISA. MTT was applied to determine the proliferation of rHpaA-immunized mouse splenocytes after stimulation of the different recombinant rPⅢ proteins. Results In the HpaA molecule there were five T/B combined epitopes: HpaA10, HpaA37, HpaA79, HpaA116 and HpaA143. All the T/B combined epitopes were successfully displayed on the surface of PⅢ protein of phage M13. The results of Western blot, ELISA and MTT confirmed that HpaA116 was the predominant antigenic epitope, both HpaA37 and HpaA79 were the efficient antigenic epitopes. However, HpaA10 and HpaA143 were identified as ineffective antigenic epitopes. Conclusion The phage display systems of T/B combined epitope peptides of H. pylori adhesin A have been successfully generated in this study. HpaA37 and HpaA79, especially HpaA116 are the efficient T/B combined antigenic epitopes in HpaA of H. pylori.