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Objective:To investigate the role of immune and inflammatory responses in the pathogenesis of gastroesophageal reflux disease (GERD).Methods:Fifteen patients with GERD who underwent biopsy and histopathology during painless gastroscopy in the digestive endoscopy center of the General Hospital of the PLA from December 2018 to September 2019 were analyzed retrospectively. They were divided into three groups: 6 cases of non erosive reflux disease (NERD), 8 cases of reflux esophagitis (RE) and 1 case of RE with high-grade intraepithelial neoplasia (RE-HIN). HE staining was used to analyze the inflammatory reaction of each group. The expression of cyclooxygenase-2(COX-2), inducible nitric oxide synthase (iNOS), interleukin (IL)-1β, myeloperoxidase (MPO), IL-4, nuclear factor kappa-B(NF-κB), IL-8, reactive oxygen species (ROS-1) were monitored by immunohistochemistry. Chi square test was used to analyze the positive rate of immune and inflammatory indexes in each group.Results:The three groups showed mild and moderate inflammatory cell infiltration, mostly lymphocyte infiltration, and basal cell hyperplasia in 3 cases. There was no significant difference between inflammatory cell infiltration, basal hyperplasia and inflammatory grade in NERD and RE( P>0.05). Immune and inflammatory factors COX-2(positive rate:NERD 4/6, RE4/8, RE-HIN 1/1), iNOS(positive rate:NERD 4/6, RE 3/8, RE-HIN 0), IL-1β(positive rate:NERD 6/6, RE 7/8, RE-HIN 1/1), MPO(positive rate:NERD 4/6, RE 7/8, RE-HIN 1/1), IL-4(positive rate:NERD 3/6, RE 4/8, RE-HIN 0), IL-8(positive rate:NERD 2/6, RE 6/8, RE-HIN 1/1), ROS-1(positive rate:NERD 3/6, RE 1/8, RE-HIN 0) and signal pathway NF- κ B (positive rate:NERD 4/6, RE 8/8, RE-HIN 1/1) were positive in three groups. The expressions of IL-1β, MPO and NF-κB were statistically significant among the three groups ( P<0.05). Conclusions:There is an inflammatory cascade mediated by immune inflammatory factors and mediators in GERD patients, and NF-κB signaling pathway is involved. It provides a basis for finding targets to block immune and inflammatory responses in the later stage to treat GERD.
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Objective:To assess the efficacy of the newly constructed system for screening, managing and monitoring congenital heart disease (CHD) in neonates of Hainan Province, thus providing references for a further promotion.Methods:Clinical data of neonatal CHD in Hainan Province from January 1, 2019 to December 31, 2021 were retrospectively analyzed, including screening, diagnosis and treatment, prognosis and follow-up.Relying on Hainan Women and Children′s Medical Center as the leading unit, a neonatal CHD screening, diagnosis, treatment, and monitoring system was established.A dual-indicator method was adopted, that was, screening staffs in Hainan Province performed CHD screening in living neonates by cardiac auscultation and pulse oximetry (POX) within 6-72 h after birth.Echocardiographic examinations for the screened living neonates were performed in the 31 authorized diagnosis institutions.Evaluations, interventions and treatment of living neonates with CHD were performed in 6 authorized tertiary hospitals.Data of screening, diagnosis, evaluation and treatment were filled in, uploaded and managed online through the neonatal CHD screening information management system.The research team of our hospital was responsible for the data management and monitoring.Results:From January 1 st, 2019 to December 31 st, 2021, there were 329 387 living neonates in Hainan Province, and 321 447 (97.59%) were screened for CHD, and the annual screening rate increased year by year.The positive rate of CHD screening was 2.50%(8 032/321 447). The rate of cardiac ultrasound examination within 1 week of CHD positive screening was 94.66%(7 603/8 032). The referral rate of severe CHD was 100.00%(154/154). The overall prevalence of CHD in neonates of Hainan Province was 3.419‰ (1 099/321 447). Atrial septal defect was the most common CHD lesion, with a proportion of 38.40%(422/1 099). The sensitivity of cardiac auscultation, POX and their combination for CHD detection were 69.15%, 33.49% and 91.90%, respectively, and the specificity were 98.36%, 99.43% and 97.81%, respectively.At the initial screening, the ratio of dual-positive of cardiac auscultation and POX in neonates with severe CHD (serious and critical CHD) was significantly higher than that of a single positive indicator ( χ2=36.502, 46.214, respectively; all P<0.001). All neonates with CHD were evaluated.Fifteen neonates with severe CHD died.From 2019 to 2021, the standardized mortality rate of children aged 0-1 years with CHD in Hainan province was 4.67/100 000 (15/321 447). Conclusions:Dual-indicator screening for CHD (cardiac auscultation plus POX) is reliable, non-invasive, and simple, which is conducive to be clinically promoted.Introducing and promoting an appropriate technology for screening, diagnosis, and evaluation of neonatal CHD are extremely significant since they may have contributed to the timely diagnosis and treatment of CHD, especially severe CHD, thus lowering the mortality.
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@#Tumor-associated macrophage promotes the progression of glioblastoma (GBM) by infiltrating into tumor tissue, yet its mechanism has not been fully elucidated.This paper aimed to investigate the mechanism of M2 macrophages in affecting the migratory capacity of GBM via secreting exosomes.Ultracentrifugation was used to extract exosomes; RNA sequencing was carried out to screen differentially expressed miRNAs; target prediction database was used to predict the possible target proteins of miRNA; Dual-luciferase reporter assay was performed to verify the interaction between miRNA and target genes; and the proliferation ability of tumor cells was detected by subcutaneous xenograft model in nude mice.Results showed that tumor-related macrophages were mainly M2 macrophages, and that exosomes secreted by M2 macrophages could promote the migration of glioma cells.Meanwhile, exosomes secreted by M2 macrophages transported miR-1260b and affected the migration of glioma cells through directly targeted AJAP1, suggesting that exosomes secreted by macrophages could affect the migration ability of GBM through transporting miR-1260b.
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Objective:To explore the value of synthetic MRI and dynamic contrast-enhanced MRI (DCE-MRI) in differentiating benign and malignant breast lesions.Methods:Totally 116 patients (female, aged 22-80 years) with suspicious breast lesions found by clinical examination in General Hospital of Ningxia Medical University from September 2020 to March 2021 were retrospectively analyzed. All patients received synthetic MRI and DCE-MRI scans, and the quantitative parameters of synthetic MRI [T 1, T 2, proton density (PD) value before enhancement of breast lesions (T 1pre, T 2pre, PD pre) and T 1, T 2, PD value after enhancement of breast lesions (T 1Gd, T 2Gd, PD Gd)] were measured. The difference before and after enhancement (?T 1, ?T 2, ?PD) and the ratio before and after enhancement(T 1ratio, T 2ratio, PD ratio) were calculated. All lesions were confirmed by biopsy or surgery. Independent sample t test was used to compare the differences of parameters between benign and malignant lesions, and parameters with statistically significant differences were analyzed by multivariate logistic regression to screen out independent predictors for the diagnosis of breast malignant lesions. The efficacy of quantitative parameters alone and in combination with DCE-MRI, was evaluated by the receiver operating characteristic (ROC) curve. Results:Of 116 patients, 27 patients were benign and 89 patients were malignant. T 2pre, PD pre, T 2Gd, PD Gd, ?T 2, T 1ratio and T 2ratio values of malignant patients were lower than those of benign lesions ( P<0.05). Multivariate logistic regression analysis showed that T 2pre (OR=0.961,95%CI 0.926-0.993, P=0.022)and PD pre(OR=0.945, 95%CI 0.908-0.979, P=0.003) were independent predictors of breast malignant lesions. The area under ROC curve (AUC) of T 2pre+PD pre in differentiating benign and malignant breast lesions was 0.77, and the accuracy, sensitivity, specificity, positive predictive value and negative predictive value were 0.84, 0.87, 0.68, 0.92 and 0.56, respectively. The AUC of T 2pre+PD pre combined with DCE-MRI in differentiating benign and malignant breast lesions was 0.95, indicating the best diagnostic efficiency, and higher than that of DCE-MRI alone (0.92)( Z=2.20, P<0.05). Conclusions:T 2pre and PD pre values of synthetic MRI can be used as reference parameters for differentiating benign and malignant breast lesions. Synthetic MRI combined with DCE-MRI can effectively improve the diagnostic efficiency of breast malignant lesions.
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The introduction of the mevalonate pathway (MVA pathway) in recombinant Escherichia coli can improve the synthesis of terpenoids. But the imbalance expression of MVA pathway genes and accumulation of intermediates inhibit cell growth and terpenoids production. In this study, each gene of MVA pathway and key genes of lycopene synthesis pathway were cloned in plasmid to express in the recombinant E. coli LYC103 with optimizing the expression of the key genes of the 2-methyl-D-erythritol-4-phosphate pathway (MEP pathway), chromosome recombinant MVA pathway and the lycopene synthesis pathway. The results showed that the overexpression of ispA, crtE, mvaK1, idi and mvaD genes did not affect the cell growth, while lycopene production increased by 13.5%, 16.5%, 17.95%, 33.7% and 61.1% respectively, indicating that these genes may be the rate-limiting steps for the synthesis of lycopene. mvaK1, mvaK2, mvaD of MVA pathway were the rate-limiting steps and were in an operon. The mvaK1, mvaK2, mvaD operon was regulated by mRS (mRNA stabilizing region) library in front of mvaK1, obtaining strain LYC104. Lycopene yield of LYC104 was doubled and cell growth was increased by 32% compared with the control strain LYC103. CRISPR-cas9 technology was used to integrate idi into chromosome at lacZ site to obtain LYC105 strain. Cell growth of LYC105 was increased by 147% and lycopene yield was increased by 2.28 times compared with that of LYC103. In this study, each gene of lycopene synthesis pathway was expressed in plasmid to certify the rate-limiting gene based on the complete MVA pathway on the chromosome. Then the rate-limiting gene was integrated in chromosome with homologous recombination to release the rate-limiting, which providing a new strategy for the construction of high-yield strains for metabolic engineering.
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Objective:To investigate risk factors for infection of biofilm-forming Klebsiella pneumonia and prognosis of patients. Methods:The clinical data of 125 patients with Klebsiella pneumoniae bloodstream infection admitted in Tianjin Medical University General Hospital from January to December 2019 were analyzed retrospectively. According to the presence of Klebsiella pneumoniae biofilm, patients were divided into biofilm positive group ( n=81) and biofim negative group ( n=44). In biofilm positive group 17 patients died (fatal group) and 64 survived (survival group) during 3-month follow-up. The antimicrobial resistance of the strains was analyzed, and multivariate logistic regression analysis was used to investigate the risk factors of biofilm-forming Klebsiella pneumoniae infection and the risk factors of death in biofilm positive group. Results:A total of 125 strains of Klebsiella pneumoniae were isolated from 125 patients, of which 81(64.80%) strains were biofilm positive. Antimicrobial resistance analysis showed that the resistance rate of biofilm positive group to aztreonam, amikacin, ciprofloxacin and levofloxacin was significantly higher than that in the biofilm negative group ( χ2=5.94, 4.03, 5.05 and 4.15, P<0.05). Multivariate logistic regression analysis showed that endotracheal intubation ( OR=3.460, 95% CI 2.890-14.445, P<0.05) and administration of immunosuppressants ( OR=6.945, 95% CI 1.160-21.567, P<0.05) within 3 months before infection were independent risk factors for biofilm-forming Klebsiella pneumoniae bloodstream infection. The use of tegacycline ( OR=4.886, 95% CI 1.123-21.263, P<0.05) within 3 months before death was independent risk factors for the death of biofilm-positive Klebsiella pneumoniae bloodstream infection. Conclusions:In order to reduce the incidence and fatality of biofilm-forming Klebsiella pneumoniae bloodstream infection, antibiotics should be used rationally and invasive procedures should be minimized.
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The authors introduced the practice of shared governance in ICU nursing management. According to the standard of magnetic hospital, a shared governance committee was established to guide and motivate the clinical front-line nurses in ICU to participate in nursing affairs management and decision-making. After three years of practice, ICU nursing management mode has achieved the goal of participating in shared governance, enhancing nurses′ perception of patient safety culture and improving the level of work engagement.
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Isoprenoids are all derived from two five-carbon building blocks called isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), which are synthesized either by the mevalonate (MVA) pathway or 2-C-methyld-D-erythritol-4-phosphate (MEP) pathway. In this study, the MVA pathway genes were integrated into the chromosome of LYC101, in which the expression of key genes in the MEP synthesis pathway and lycopene synthesis pathway were optimized by artificial regulatory parts, to further improve the production of isoprenoids in Escherichia coli. The plasmids pALV23 and pALV145 were screened from a plasmid library that constructed by using the RBS library to link the genes of the MVA pathway, which greatly increased the production of β-carotene. The effects of plasmids pALV23 and pALV145 on the lycopene production in low and high lycopene production strain, LYC001 and LYC101, were compared, respectively. The production of lycopene was increased by plasmids pALV23 and pALV145 in both strains. In high lycopene production strain LYC101, pALV23 produced more lycopene than pALV145. Then, the MVA gene together of promoter of pALV23 was integrated into the chromosome of LYC101 at poxB site using method of homologous recombination helped by CRISPR-Cas9 system, resulted in genetically stable strain, LYC102. The yield of lycopene of LYC102 was 40.9 mg/g DCW, 1.19-folds higher than that of LYC101, and 20% more than that of LYC101 with pALV23. Simultaneous expression of MVA pathway and MEP pathway in recombinant E. coli can effectively increase the yield of terpenoids. In this study, a plasmid-free, genetically stable, high-yielding lycopene strain was constructed, which could be used for industrialization. Also, the platform strain can be used for the synthesis of other terpenoids.
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Chromosomes, Bacterial , Escherichia coli , Lycopene , Mevalonic Acid , beta CaroteneABSTRACT
Objective To study the detection of cytomegalovirus (CMV)-DNA in different kinds of blood and urine sample .Methods The CMV-DNA loads in 3 different kinds of blood sample from 52 patients and 3 different kinds of urine sample from 85 patients were detected by real-time fluorescence quantitative polymer-ase chain reaction(FQ-PCR) .The differences in CMV-DNA detection rate and virus loads were compared a-mong different kinds of blood and urine samples .Results The CMV-DNA detection rates in serum ,whole blood ,plasma and peripheral blood mononuclear cell (PBMC) from 52 patients were 48 .08％ ,71 .15％ ,57 .69％ and 69 .23％ respectively .The CMV-DNA detection rates of whole blood and PBMC were higher ,the difference was statistically significant (P<0 .05) .The quantitative results of PBMC was higher .The CMV-DNA detec-tion rates of mixed urine ,urine supernatant and urine sediment from 85 patients were 72 .94％ ,62 .35％ and 84 .71％ respectively ,the CMV-DNA detection rate of urine sediment was higher ,while the quantitative re-sults of mixed urine was higher .Conclusion The CMV-DNA detection results of blood and urine have large difference ,therefore using the same kind of sample for conducting detection has an important clinical signifi-cance in the clinical diagnosis ,treatment and monitoring of CM V infection .
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Objective To investigate the clinical efficacy and application value of submucosal tunneling endoscopic resection(STER)for upper gastrointestinal submucosal tumors(SMTs). Methods A retrospective analysis was performed on the endoscopic and clinical data of 44 cases with SMTs who received STER from January 2015 to June 2016 in Chinese PLA General Hospital. Results The rate of complete resection was 88.6%(39/44). The operating time was 60.1±30.6 min. The hospitalization time was 10.1± 3.3 days. The rate of complications was 6.8%(3/44). The diagnosis of SMTs by pathology and endoscopic ultrasonography(EUS),the size of SMTs measured by EUS and ruler after STER,and the growing direction judged by EUS and CT were consistent. Conclusion STER for SMTs has a higher complete resection rate, shorter operating time and hospitalization time, and fewer complications. EUS combined with CT is an effective method for preoperative evaluation.
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Objective To study effects of shuganjieyu capsule of plasma glucose and lipid metabolism and sleep and body weight in patients with depression.Methods 90 patients of depression who received therapy from July 2013 to July 2016 in our hospital were selected.According to random number table, those patients were divided into the observation group ( n =45 ) and the control group ( n =45 ) .The control group was treated with citalopram, while the observation group was treated with shuganjieyu capsule, then glucose and lipid metabolism, sleep, weight and clinical curative effect were compared.Results After treatment, the fasting blood glucose (FBG), postprandial 2h blood glucose (2 hPG), triacylglycerol (TG), total cholesterol (TC) in the observation group were lower than the control group(all P<0.05), the sleep state self rating scale (SRSS) and the Pittsburgh sleep quality index ( PSQI) in the observation group were lower than the control group ( all P <0.05 ) .The body weight and body mass index in the observation group were lower than the control group (all P<0.05).The total effective rate in the observation group 95.56% (43/45)was higher than the control group 77.78% (35/45) (P<0.05); the incidence of adverse reactions in observation group 20.00% (9/45) was lower the control group 46.67% (21/45) ( P <0.05).Conclusion Shuganjieyu capsule is well for depression, which can conducive to maintaining glucose and lipid metabolism, weight stability, and improve sleep quality and clinical symptoms, and high security,it's worthy of application and promotion.
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Objective To explore the feasibility of endothelial function of dorsal artery of foot in patients with type 2 diabetes(T2DM) by high frequency ultrasound combined with warm bath test.MethodsThirty-five patients with T2DM and thirty normal people were collected,all subjects were examined by high frequency ultrasound.Diameter of brachial artery in baseline and after reactive hyperaemia were detected;Diameter of dorsal artery of foot in baseline and after the foot immersed in 40℃ warm water for 5 minutes were acquired.Flow mediated dilatation of dorsalis pedis artery(FMDDPA) and flow mediated dilatation of brachial artery(FMDBA) were calculated and compared.Multiple stepwise regression analysis was used to examine the correlation between FMDDPA and hemoglobin A1c(HbAlc). Results The FMDDPA and FMDBA were decreased in patients with T2DM (P<0.01).The FMDDPA and FMDBA were correlated significantly(r=0.864,P<0.01).In stepwise regression analysis,HbA1c is the most affecting factor for FMDDPA (R2=0.321,P<0.01).Conclusions Ultrasound combined with warm bath test can be used to detect the change of endothelial function of dorsal artery of foot in patients with T2DM,which have a certain clinical application value in endothelial function of terminal limb artery.
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Objective To explore the curative effect of quetiapine combined with paroxetine in the treatment of bipolar depression,and to provide reference for the choice of clinical treatment.Methods According to the digital table,96 patients with bipolar disorder were randomly divided into observation group and control group,48 cases in each group.The control group was treated with quetiapine orally,the observation group was treated with quetiapine combined with paroxetine.After 6 weeks of treatment,the clinical curative effect,the Montgomery Asberg Depression Rating Scale and the Beck pull fan Sen Mania Rating table score before and after treatment were compared between the two groups.At the same time,the adverse reactions of the two groups were compared.Results After 6 weeks of treatment,the total effective rate of the observation group was 70.83％,which was significantly higher than 50.00％ in the control group,the difference was statistically significant(x2 =4.356,P =0.037).After 6 weeks of treatment,the Montgomery Asberg Depression Rating quantity score in the observation group was (14.01 ± 2.95) points,which was significantly lower than (18.07 ± 3.05)points in the control group,the difference was statistically significant (t =5.527,P =0.036).The Baker-pull fan Sen Mania Rating score before and after treatment between the two groups had no statistically significant differences.There were no serious adverse reactions in the two groups.Conclusion Quetiapine combined with paroxetine can effectively inhibit bipolar depressive episodes,avoid manic episodes,has good clinical efficacy and high safety,it is worthy of popularization and application.
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Objective To determine the optimal cutoff value of carcinoembryonic antigen (CEA) and amylase in cyst fluid for mucinous cystic neoplasm,to explore the difference of cutoff value of CEA between Chinese and western populations and the diagnostic accuracy of CEA combined with amylase for mucinous pancreatic cysts.Methods A total of 116 patients received EUS-FNA in Chinese PLA General Hospital from April 2014 to May 2016 with cyst fluid for biochemical and histological examinations and biopsy for pathological examination.The optimal value of amylase and CEA for mucinous pancreatic cysts were obtained from receiver operator characteristics curve of CEA and amylase.The accuracy,sensitivity and specificity of CEA,amylase and combination of both were calculated.Results Diagnosis of 70 patients were confirmed by surgery,forceps or cytology.There were 32 cases of non-mucinous pancreatic cysts including 6 pseudocysts and 26 serous cystadenoma.There were 38 cases of mucinous pancreatic cysts including 31 mucinous cystic neoplasm and 7 intraductal papillary mucinous neoplasm.The optimal cutoff of CEA was 72.35 ng/mL.The accuracy,sensitivity and specificity were 84.3％,81.6％ and 87.5％ respectively.The accuracy,sensitivity and specificity for mucinous cystic neoplasm were 80.0％,71.1％ and 90.6％ respectively when adopting CEA ＞ 192 ng/mL.The optimal cutoff of amylase was 461.70 IU/L,and accuracy,sensitivity and specificity were 57.1％,68.4％,43.8％ respectively.Combination of CEA＞72.35 ng/mL and amylase＜461.7 IU/L yielded higher accuracy (85.7％) and specificity (93.8％) with lower sensitivity (78.9％).Conclusion CEA can be used in the differential diagnosis of mucinous and nonmucinous pancreatic cysts.The optimal cutoff of CEA in Chinese was lower than that in western populations.And the combined analysis of CEA and amylase could increase the diagnostic accuracy.However,in order to confirm this conclusion,a study of larger scale is needed.
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OBJECTIVE:To establish the quality standard for Chaige ganmao mixture. METHODS:TLC was conducted to iden-tify the Scutellaria baicalensis and Paeonia lactiflora;HPLC was conducted to determine the content of puerarin. The column was Zorbax Eclipse XDB-C18 with mobile phase of methanol-water(25:75,V/V)at flow rate of 1.0 ml/min,detection wavelength was 250 nm,and column temperature was 25℃,and the injection volume was 5μl. RESULTS:TLC showed clear spots and good sepa-ration. The linear range of puerarin was 18-198 μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were low-er than 2%,recovery was 99.04%-99.75%(RSD=0.28%,n=6). CONCLUSIONS:The standard can be used for the quality con-trol of Chaige ganmao mixture.
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Patients with metastatic prostate cancer are typically managed with androgen deprivation therapy. Most patients initially respond to treatment, but many eventually develop castration-resistant prostate cancer. Neuroendocrine prostate cancer (NEPC) is a highly aggressive subtype of castration-resistant prostate cancer, which often results from neuroendocrine differentiation of prostate cancer cells. NEPC has a dismal outcome with an average survival of less than 1 year and exhibits less response to radiochemotherapy. At least 25%of patients with lethal castration-resistant prostate cancer are predicted to eventually develop this type of highly-aggres-sive NEPC. However, research on the molecular mechanism of NEPC is limited;thus, further studies are needed to explore the develop-ment and application of anticancer drugs for NEPC.
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ObjectiveTo observe the effect ofTong Du acupuncture-moxibustion, conventional acupuncture, and medication on the bone marrow stem cells (BMSCs) mobilization markers CD34+and CD133+, neurological deficit score in patients with ischemic stroke.Method Totally 120 eligible subjects with ischemic stroke were randomized into aTong Du acupuncture-moxibustion group (group A), a conventional acupuncture group (group B), and a medication group (group C), 40 in each group. In addition to the conventional medication given to all three groups, patients in group A also received moxibustion at Baihui (GV20), bloodletting at Dazhui (GV14), and acupuncture at Fengchi (GB20), Quchi (LI11), Hegu (LI4), Zusanli (ST36), and Sanyinjiao (SP6); patients in group B also received acupuncture at Fengchi, Quchi, Hegu, Zusanli, and Sanyinjiao; group C received patent Chinese medicine Tong Xin Luo capsules. The intervention lasted 14 d in all three groups. The CD34+and CD133+and neurological deficit score were evaluated and compared before and after intervention.Result The three treatment methods all significantly increased the level of CD34 + and CD133+in peripheral blood (P0.05); the three treatment methods all improved the neurological deficit score to various extent, and the result in group A was significantly different from that in group B and C (P<0.05,P<0.01), and there was also a significant difference between group B and C (P<0.05).ConclusionsTong Du acupuncture-moxibustion, conventional acupuncture, and medication can influence the BMSCs and neurological deficit to different extent; the effect ofTong Du acupuncture-moxibustion on the BMSCs and neurological deficit is more significant than that of the conventional acupuncture and medication, suggesting thatTong Du acupuncture-moxibustion method should have advantage in treating cerebral infarction.
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BACKGROUND:Tissue engineering provides new ideas and approaches for repair of cartilage defects. OBJECTIVE:To develop a complete set of solutions for construction of tissue engineered cartilagein vitro, with chondrocytes as seed cels and cross-linked sodium hyaluronate as scaffold materials. METHODS: New Zealand rabbit articular chondrocytes were isolated, counted, and then cultured and passaged to prepare cellsuspension. Toluidine blue staining, RT-PCR and immunocytochemistry were exerted to evaluate the cultured cels. Chondrocytes were seeded and co-cultured with cross-linked sodium hyaluronate scaffold for 21 days. Then, RNA was isolated for RT-PCR, and frozen sections were prepared for morphological observation and immunohistochemistry study. RESULTS AND CONCLUSION:The chondrocytes could adhere to the cross-linked sodium hyaluronate scaffold and aggregate, growing between fibers or adhering to the scaffold in a monolayer manner. The transcripts of cartilage specific aggrecan gene and colagen type II alpha 1 gene and cartilage specific protein colagen type II were expressed in cel-scaffold complexes to maintain the phenotype of chondrocytes. Cel-scaffold complexes co-cultured in vitro can form cartilage extracelular matrix, by which tissue engineered cartilage is expected to be obtained.
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An HPLC method has been developed to determine polydatin in giant knotweed rhizome. In order to systematically validate the method, specificity, precision, linearity of reference solution and test solution, repeatability, reproducibility, accuracy, stability and robustness were measured. In the robustness test, a one-variable-at-a-time procedure was applied to evaluate the influence of slight variations in method factors, including the flow rate, the column temperature, the extraction time, and etc., on the assay result of polydatin. No significant differences were found when the process parameters changed during the experimental domain. And system suitability test limits were defined based on the robustness test. Results showed that the developed method was accurate, reproducible and robust.
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Objective To study the expression of Gli1 protein and vascular endothelial growth factor (VEGF)-C in papillary thyroid carcinoma and their relationship with cervical lymph node metastasis.Methods The expression of Gli1 protein and VEGF-C were investigated by immunohistochemical EnVision method in 67 cases of papillary thyroid carcinoma and 32 cases of nodular goiter pathological specimen tissues.Nascent microvascular and micro-lymphatic of all pathological speimen tissues were examined by immunohistochemical monoclonal antibody CD34 and D2-40 staining assay respectively,and the microvascular density (MVD) and micro-lymphatic vessel density (MLVD) were calculated.The relationship between the positive expression of Gli1 protein and MVD,between the positive expression of VEGF-C and MLVD,and their relationship with cervical lymph node metastasis were analyzed.Results The positive rates of Gli1 protein,VEGF-C and MVD,MLVD were significantly higher in papillary thyroid carcinoma than those in nodular goiter [67.16％(45/67) vs.43.75％(14/32),70.15％(47/67) vs.31.25％(10/32),23.14 ± 2.06 vs.2.14 ± 0.31,13.36 ± 1.32 vs.3.53 ± 0.65,P ＜ 0.05].The positive expression of Gli1 protein was related to the patient' s age and the tumor size (P ＜ 0.05),while the positive expression of VEGF-C was not related to the patient' s age and the tumor size (P ＞ 0.05).The positive expression of Gli1 protein and VEGFC were significantly higher in TNM stage Ⅲ-Ⅳ of papillary thyroid carcinoma than those in TNM stage Ⅰ-Ⅱ (P ＜ 0.01) and also was significantly higher with cervical lymph node metastasis than without cervical lymph node metastasis (P ＜ 0.01).MVD and MLVD were significantly higher with cervical lymph node metastasis than without cervical lymph node metastasis (P ＜ 0.05).The positive expression of Gli1 protein was positively correlated with MVD (r =0.784,P＜ 0.01),the positive expression of VEGF-C was positively correlated with MLVD (r =0.529,P ＜ 0.01),the positive expression of Gli 1 protein was positively correlated with the positive expression of VEGF-C (r =0.586,P ＜0.01).Conclusions Gli1 protein which may participate in the formation of nascent microvascular is abnormally activated by the Hedgehog signaling pathway to express,VEGF-C which may be participate in the formation of nascent micro-lymphatic is mediated and started by the MAPK signaling pathways to express.Expressions of Gli1 protein is positively correlated with expressions of VEGF-C,therefore the Hedgehog signaling pathway may be associated with the MAPK signal pathway.Suppressing nascent microvascular and micro-lymphatic may become new target to blockingup papillary thyroid carcinoma cervical lymph node metastasis.