Postmortem Interval Estimation Using Protein Chip Technology Combined with Multivariate Analysis Methods / 法医学杂志

OBJECTIVES@#To estimate postmortem interval (PMI) by analyzing the protein changes in skeletal muscle tissues with the protein chip technology combined with multivariate analysis methods.@*METHODS@#Rats were sacrificed for cervical dislocation and placed at 16 ℃. Water-soluble proteins in skeletal muscles were extracted at 10 time points (0 d, 1 d, 2 d, 3 d, 4 d, 5 d, 6 d, 7 d, 8 d and 9 d) after death. Protein expression profile data with relative molecular mass of 14 000-230 000 were obtained. Principal component analysis (PCA) and orthogonal partial least squares (OPLS) were used for data analysis. Fisher discriminant model and back propagation (BP) neural network model were constructed to classify and preliminarily estimate the PMI. In addition, the protein expression profiles data of human skeletal muscles at different time points after death were collected, and the relationship between them and PMI was analyzed by heat map and cluster analysis.@*RESULTS@#The protein peak of rat skeletal muscle changed with PMI. The result of PCA combined with OPLS discriminant analysis showed statistical significance in groups with different time points (P<0.05) except 6 d, 7 d and 8 d after death. By Fisher discriminant analysis, the accuracy of internal cross-validation was 71.4% and the accuracy of external validation was 66.7%. The BP neural network model classification and preliminary estimation results showed the accuracy of internal cross-validation was 98.2%, and the accuracy of external validation was 95.8%. There was a significant difference in protein expression between 4 d and 25 h after death by the cluster analysis of the human skeletal muscle samples.@*CONCLUSIONS@#The protein chip technology can quickly, accurately and repeatedly obtain water-soluble protein expression profiles in rats' and human skeletal muscles with the relative molecular mass of 14 000-230 000 at different time points postmortem. The establishment of multiple PMI estimation models based on multivariate analysis can provide a new idea and method for PMI estimation.

Clinicopathological features and prognostic factors of patients with lung metastasis of stage Ⅰa~Ⅲb cervical cancer / 中华肿瘤杂志

Objective: To investigate the clinicopathological features and prognostic factors of lung metastasis in patients with cervical cancer after treatment. Methods: The clinicopathological data of 191 patients with lung metastasis of stage Ⅰa-Ⅲb cervical cancer (FIGO 2009 stage) treated in Sichuan Cancer Hospital from January 2007 to December 2020 were analyzed retrospectively. Kaplan Meier method and Log rank test were used for survival analysis, and Cox regression model was used for prognostic factors analysis. Results: Among 191 patients with lung metastasis of cervical cancer, pulmonary metastasis was found in 134 patients (70.2%) during follow-up examination, and 57 patients (29.8%) had clinical symptoms (cough, chest pain, shortness of breath, hemoptysis, and fever). The time from the initial treatment of cervical cancer to the discovery of lung metastasis was 1-144 months in the whole group, with a median time of 19 months. Univariate analysis of the prognosis of lung metastasis after treatment of cervical cancer showed that the diameter of cervical tumor, lymph node metastasis, positive surgical margin, disease-free interval after treatment of cervical cancer, whether it is accompanied by other metastasis, the number, location and maximum diameter of lung metastasis, and the treatment method after lung metastasis are related to the prognosis of patients with lung metastasis of cervical cancer. Multivariate analysis showed that the number of lung metastases and other site metastases in addition to lung metastases were independent factors affecting the prognosis of patients with lung metastases of cervical cancer (P<0.05). Conclusions: For patients with cervical cancer, attention should be paid to chest CT examination during follow-up to guard against the possibility of lung metastasis after treatment. Besides lung metastasis, other site metastasis and the number of lung metastasis are independent factors affecting the prognosis of patients with lung metastasis of cervical cancer. For patients with lung metastasis after treatment of cervical cancer, surgical treatment is an effective treatment. It is necessary to strictly grasp the surgical indications, and some patients can achieve long-term survival. For patients with lung metastasis of cervical cancer who are not suitable for resection of lung metastasis, the remedial treatment of chemotherapy with or without radiotherapy is still a recommended choice.

Systematic review and Meta-analysis of efficacy and safety of Kushen Gelatum combined with antibiotics in treatment of bacterial vaginosis / 中国中药杂志

This study aims to systematically evaluate the clinical efficacy and safety of Kushen Gelatum combined with antibiotics for treating bacterial vaginosis. The randomized controlled trial(RCT) of Kushen Gelatum for treating bacterial vaginosis were retrieved from CNKI, Wanfang, VIP, SinoMed, PubMed, and Cochrane Library with the time interval from inception to January 2023. Data were extracted from the included RCT by 2 investigators, including the sample size, characteristics of patients, interventions and controls, outcome indicators, and adverse effects. The Cochrane collaboration network's bias risk assessment tool was used for methodolo-gical quality evaluation of the included trials. RevMan 5.4 was employed to perform the Meta-analysis. A total of 19 RCTs were inclu-ded, involving 1 980 patients with bacterial vaginosis. Meta-analysis showed that, compared with nitroimidazoles alone, Kushen Gelatum + nitroimidazoles improved the total response rates in terms of clinical symptoms and laboratory tests(RR=1.24, 95%CI[1.13, 1.36], P<0.000 01), laboratory tests(RR=1.16, 95%CI[1.06, 1.26], P=0.000 9), and clinical symptoms(RR=1.26, 95%CI[1.08, 1.46], P=0.003), and reduced the leukocyte esterase positive rate(RR=0.29, 95%CI[0.17, 0.48], P<0.000 01) and the recurrence rate(RR=0.37, 95%CI[0.23, 0.58], P<0.000 1). Compared with lincomycin antibiotics(clindamycin) alone, Kushen Gelatum + lincomycin antibiotics(clindamycin) improved the total response rates in terms of clinical symptoms and laboratory tests(RR=1.18, 95%CI[1.06, 1.31], P=0.003) and laboratory tests(RR=1.27, 95%CI[1.04, 1.54], P=0.02), reduced the recurrence rate(RR=0.20, 95%CI[0.05, 0.75], P=0.02), and shortened the time to relief of burning sensation(MD=-1.70, 95%CI[-2.15,-1.26], P<0.000 01), vaginal itching(MD=-0.82, 95%CI[-1.30,-0.34], P=0.000 8), and abnormal leucorrhea(MD=-1.52, 95%CI[-1.98,-1.06], P<0.000 01). Compared with nitroimidazoles + probiotics, Kushen Gelatum + nitroimidazoles + probiotics improved the total response rate in terms of clinical symptoms and laboratory tests(RR=1.18, 95%CI[1.02, 1.36], P=0.03) and reduced the recurrence rate(RR=0.27, 95%CI[0.09, 0.76], P=0.01). Kushen Gelatum combined with antibiotics demonstrates a potential therapeutic effect on bacterial vaginosis, whereas the number and quality of the relevant clinical studies remain to be improved. The process of clinical trial should be standardized to improve the quality of evidence, so as to provide strong evidence to guide the application of Kushen Gelatum in clinical practice.

Effect of baicalin modulation of TLR4/MAPK/NF-ΚB signaling pathway on renal fibrosis in rats with diabetic nephropathy / 中国药理学通报

Aim To investigate the protective effect of baicalin on renal fibrosis in rats with diabetic nephrop- III (Col- III) athy (DN) and to investigate its mechanism of action. Methods A rat model of diabetic nephropathy was constructed. The rats were randomly divided into control group, model group, baicalin low dose group, baicalin medium dose group, baicalin high dose group and metformin group, with 10 rats in each group. Except for the control group, all rats in each group were fed with streptozotocin 65 mg • kg -

Analysis of prognostic factors of extranodal NK/T-cell lymphoma treated with pegaspargase/L-asparaginase: a multicenter retrospective study / 中华血液学杂志

Objective: To explore the prognostic factors of extracellular NK/T cell lymphoma (ENKTL) treated with pegaspargase/L-asparaginase. Methods: The clinical data of 656 ENKTL patients diagnosed at 11 medical centers in the Huaihai Lymphoma Working Group from March 2014 to April 2021 were retrospectively analyzed. The patients were randomly divided into two groups: a training set (460 cases) and a validation set (196 cases) at 7∶3, and the prognostic factors of the patients were analyzed. A prognostic scoring system was established, and the predictive performance of different models was compared. Results: Patients' median age was 46 (34, 57) years, with 456 males (69.5% ) and 561 nasal involvement (85.5% ). 203 patients (30.9% ) received a chemotherapy regimen based on L-asparaginase combined with anthracyclines, and the 5-year overall survival rate of patients treated with P-GEMOX regimen (pegaspargase+gemcitabine+oxaliplatin) was better than those treated with SMILE regimen (methotrexate+dexamethasone+cyclophosphamide+L-asparaginase+etoposide) (85.9% vs 63.8% ; P=0.004). The results of multivariate analysis showed that gender, CA stage, the Eastern Cooperative Oncology Group performance status (ECOG PS) score, HGB, and EB virus DNA were independent influencing factors for the prognosis of ENKTL patients (P<0.05). In this study, the predictive performance of the prognostic factors is superior to the international prognostic index, Korean prognostic index, and prognostic index of natural killer lymphoma. Conclusion: Gender, CA stage, ECOG PS score, HGB, and EB virus DNA are prognostic factors for ENKTL patients treated with pegaspargase/L-asparaginase.

Noradrenaline modulates the spontaneous firing activities of Purkinje cells via α2-adrenergic receptor in mouse cerebellar cortex / 生理学报

Cerebellar Purkinje cells (PCs) exhibit two types of discharge activities: simple spike (SS) and complex spike (CS). Previous studies found that noradrenaline (NA) can inhibit CS and bidirectionally regulate SS, but the enhancement of NA on SS is overwhelmed by the strong inhibition of excitatory molecular layer interneurons. However, the mechanism underlying the effect of NA on SS discharge frequency is not clear. Therefore, in the present study, we examined the mechanism underlying the increasing effect of NA on SS firing of PC in mouse cerebellar cortex in vivo and in cerebellar slice by cell-attached and whole-cell recording technique and pharmacological methods. GABAA receptor was blocked by 100 µmol/L picrotoxin in the whole process. In vivo results showed that NA significantly reduced the number of spikelets of spontaneous CS and enhanced the discharge frequency of SS, but did not affect the discharge frequency of CS. In vitro experiments showed that NA reduced the number of CS spikelets and after hyperpolarization potential (AHP) induced by electrical stimulation, and increased the discharge frequency of SS. NA also reduced the amplitude of excitatory postsynaptic current (EPSC) of parallel fiber (PF)-PC and significantly increased the paired-pulse ratio (PPR). Application of yohimbine, an antagonist of α2-adrenergic receptor (AR), completely eliminated the enhancing effect of NA on SS. The α2-AR agonist, UK14304, also increased the frequency of SS. The β-AR blocker, propranolol, did not affect the effects of NA on PC. These results suggest that in the absence of GABAA receptors, NA could attenuate the synaptic transmission of climbing fiber (CF)-PC via activating α2-AR, inhibit CS activity and reduce AHP, thus enhancing the SS discharge frequency of PC. This result suggests that NA neurons of locus coeruleus can finely regulate PC signal output by regulating CF-PC synaptic transmission.

Acupoint selection rules of acupuncture and moxibustion for stroke-related pneumonia / 中国针灸

Using data mining technology, the rules of acupoint selection of acupuncture and moxibustion were explored in treatment of stroke-related pneumonia. The clinical articles of acupuncture and moxibustion in treatment of stroke-related pneumonia were retrieved from CNKI, SinoMed, Wanfang and VIP databases from their inception through to January l 2021, and then, the acupuncture-moxibustion prescription database was set up for stroke-related pneumonia. SPSS Modeler 18.0 Apriori algorithm was adopted to analyze the association rules of acupoints and draw complex network diagrams. SPSS26.0 was used in clustering analysis of acupoints. Finally, a total of 44 articles were included, with 51 acupoint prescriptions and 82 acupoints extracted. The total frequency of acupoints was 340 times. The high-frequency acupoints in treatment with acupuncture and moxibustion for stroke-related pneumonia were Feishu (BL 13), Fenglong (ST 40), Hegu (LI 4), etc. These acupoints were mainly distributed on the limbs and back and mostly from yang meridians. Of these extracted acupoints, the five-shu points, convergent points and back-shu points were selected specially. Regarding acupoint combination, the association of Quchi (LI 11) had the highest support with Hegu (LI 4) and Zusanli (ST 36). The core prescription of acupuncture-moxibustion treatment for stroke-related pneumonia could be composed of Quchi (LI 11), Hegu (LI 4), Zusanli (ST 36), Fenglong (ST 40) and Taichong (LR 3).

Research on the Identification Model of Medical Damage / 法医学杂志

OBJECTIVES@#To understand the perceptions of doctors, patients and forensic examiners on the current situation of medical disputes and medical damage identification in China, and to explore the medical damage identification model that is more conducive for the resolution of medical disputes.@*METHODS@#A questionnaire was designed, and in-service clinicians, forensic examiners and inpatients in Sichuan Province and Chongqing City were randomly selected from April to November 2019. SPSS 22.0 software was used to analyze the data of various survey results.@*RESULTS@#Compared with patients (24.92%), doctors (61.72%) believed that the current doctor-patient relationship was more tense than before; both doctors and patients were more inclined to choose voluntary consultation and people's mediation to resolve medical disputes; forensic examiners have the highest level of cognition of medical and health-related laws and regulations, followed by doctors and patients; 66.72% of doctors and 78.41% of patients believed that medical damage identification was necessary, and they were more inclined to entrust forensic identification institutions; different groups all believed that forensic examiners and doctors should participate in the identification together, 80.94% of doctors believed that the appraisal institutions should be responsible for the forensic opinion, not the appraiser.@*CONCLUSIONS@#It is suggested that the Medical Association identification and forensic identification should learn from each other and formulate basic unified rules for the identification of medical damage. It is suggested to standardize the behavior of medical damage forensic identification institutions and appraisers, to improve their own appraisal level, actively invite clinical medical experts for consultation in identification, and promote the standardized, scientization of forensic identification.

Comparison of Medical Dispute Resolution Mechanisms in China and Abroad / 法医学杂志

Medical disputes are one of the common problems concerned by the whole world. All countries and regions have established their own medical dispute resolution mechanisms, in accordance with their own national conditions. Medical dispute identification opinions, as one of the important bases for identifying the responsibilities of both doctors and patients, play a pivotal role in the process of dispute settlement. A reasonable medical dispute resolution mechanism and standardized medical dispute identification model can help resolve disputes flexibly and reduce the conflict between doctors and patients. This paper briefly compares the medical dispute resolution mechanism and identification mode of China and several other representative countries (the United States, Britain, France, Germany, Italy, Japan, etc.), and discusses their respective characteristics and shortcomings, to bring some enlightenment to the medical dispute resolution and identification in our country.

Osteomodulin (OMD) as A Potential Prognostic Marker of GastricCancer and A New Immunotherapy Target / 中国生物化学与分子生物学报

Preoperative detection of biomarkers that can predict postoperative survival of gastric cancer patients has important implications for surgical procedures, postoperative chemoradiotherapy and followup. Using multi-center cancer database and online analysis and verifying by qRT-PCR and Western blotting, we found that Osteomodulin (OMD) was highly expressed in gastric cancer tissues (P =0. 015) and could affect the survival of gastric cancer patients (P < 0. 001) and can be detected preoperatively to evaluate the prognosis of gastric cancer patients. The mRNA expression of OMD was significantly correlated with age (P = 0. 034), Lauren typing (P < 0. 001) and clinical stage (P =0. 001) of gastric cancer patients. It also associated with a variety of immune cells (dendritic cellsresting, eosinophils) and the immune checkpoint regulator ENTPD1 (rho = 0. 634, P < 0. 001) and chemokine CXCL12 (rho = 0. 625, P < 0. 001), which affects the occurrence and development of gastriccancer through the immune microenvironment. Therefore, OMD may become a clinically feasible prognostic biomarker of gastric cancer and a new target for immunotherapy.

Soy isoflavones inhibit inflammation and apoptosis of hippocampal neurons induced by Ap, -42 through NF-kB p65 signaling pathway / 中国药理学通报

Aim To explore the effeet of soy isofla- vones (SI) on p-amyloid 1 -42 ( Ap, _42 ) -induced hippocampal neuroinflammation and neuronal apoptosis and the underlying mechanism.Methods The prima¬ry hippocampal neurons cultured in vitro were divided into control group (control), Ap,_42 treatment group f model) , SI low-dose group ( Sl-L, 10 mg • L 1 ) , and SI medium-dose group (SI-M, 20 mg • L_l ) and SI high-dose group (SI-H, 40 mg • L 1 ).The model group was treated with 30 (xmol • L"1 Ap, _42 for 48 h; the SI-L, SI-M and SI-H groups were treated with SI for 2 hours, and Ap,_42 was treated for 48 h; the con¬trol group was routinely cultured for 48 h.MTT method was used to detect the survival rate of hippocampal neurons; TUNEL staining was used to detect the apop¬tosis rate of hippocampal neurons; Western blot was used to detect COX-2, TNF-a, NF-kB p65 , P-NF-kB p65, Bcl-2 and caspase-3 protein expression levels.Results Compared with the control group, the surviv¬ al rate of hippocampal neurons was significantly re- duced (P <0.01) , and the apoptotie rate significantly increased (P<0.01).COX-2, TNF-a, p-NF-KB p65 , caspase-3 protein expressions markedly increased (P <0.05 or P <0.01 ) , and the expression of Bcl-2 protein significantly decreased in the model group ( P <0.01 ).Compared with the model group, the surviv¬al rate of hippocampal neurons, Bcl-2 protein in-creased, and the apoptotic rate, the expression of COX-2, TNF-a, p-NF-KB p65 , caspase-3 protein de¬creased (P < 0.05 or P < 0.01 ) in SI each dose group.Conclusion SI can reduce the hippocampal neuroinflammation and neuronal apoptosis induced by APi _42 by inhibiting the activation of NF-kB p65 signa¬ling pathway.

Relationship between Wound Age and Serum Marker Metabolites of Rats Skin Incised Wound / 法医学杂志

Objective To observe the metabolomics changes of serum after skin incision of rats and to determine the wound age of skin incision. Methods A rat skin incision model was established, 21 SD rats were divided into 1 h, 2 h, 4 h, 8 h, 16 h, 24 h after skin incision groups and the control group, then blood was taken from rats in the experimental groups at the corresponding time points after injury, and taken from the control group directly. Gas chromatography-mass spectrometry （GC-MS） technology was used to detect serum metabolites and screen marker metabolites, then orthogonal partial least square-discriminant analysis （OPLS-DA） model was used to establish a regression model for the relationship between marker metabolite content and wound age to determine wound age of skin. Results GC-MS was used to detect the serum collected, and 21 marker metabolites were obtained through initial screening, and 4 marker metabolites were further analyzed and screened using multivariate statistical analysis methods. There was no correspondence between the change rule of the serum content and wound age, therefore it cannot be used directly to determine wound age. OPLS model could be used to obtain regression models of the content and wound age of 21 marker metabolites and 4 marker metabolites, both of which can determine wound age, but the prediction accuracy of the regression model of 21 marker metabolites was significantly higher. Conclusion Using metabolomics to establish a regression model of the metabolite content and wound age has the potential to be applied to skin incision wound age determination.

Quchi (LI 11) for treatment of allergic diseases based on coincidence of key therapeutic characteristics of acupoints and pathogenesis of allergic diseases / 中国针灸

According to the etiology, allergic diseases are related to wind and heat; according to the pathogenesis, most of allergic diseases are

Analysis on specific effect of acupoints at the upper arms in treatment of scrofula and goiter / 中国针灸

Through analyzing the indication distribution of the different acupoints located at the upper limbs recorded in

Erratum to: The treatment effect of novel hGHRH homodimer to male infertility hamster

The authors note that on page 637 (Author Name), author affiliation of Tao Liu “Tao Liu⁶” should instead appear as “Tao Liu⁵.”

Exploration into the simplex reinforcing-reducing manipulation of acupuncture / 中国针灸

Four controversial types of simplex reinforcing-reducing manipulation of acupuncture and their possible meanings were summarized to explore several key elements of reinforcing-reducing manipulation of acupuncture, in addition, the simplex reinforcing-reducing manipulation of acupuncture was classified by single factor. It is concluded that the definition of simplex reinforcing-reducing manipulation of acupuncture should try not to include other non-manipulative elements. According to single factor, it can be divided into: needle-oriented reinforcing-reducing manipulation, twisting reinforcing-reducing manipulation, lifting and interpolating reinforcing-reducing manipulation, fast and slow reinforcing-reducing manipulation, breathing reinforcing-reducing manipulation, opening and closing reinforcing-reducing manipulation. In addition, after considering the effect and principle of number reinforcing-reducing manipulation, it can be considered.

The treatment effect of novel hGHRH homodimer to male infertility hamster

Extra-hypothalamic growth hormone-releasing hormone (GHRH) plays an important role in reproduction. To study the treatment effect of Grin (a novel hGHRH homodimer), the infertility models of 85 male Chinese hamsters were established by intraperitoneally injecting 20 mg/kg of cyclophosphamide once in a week for 5 weeks and the treatment with Grin or human menopausal gonadotropin (hMG) as positive control was evaluated by performing a 3-week mating experiment. 2–8 mg/kg of Grin and 200 U/kg of hMG showed similar effect and different pathological characteristics. Compared to the single cyclophosphamide group (0%), the pregnancy rates (H-, M-, L-Grin 26.7, 30.8, 31.3%, and hMG 31.3%) showed significant difference, but there was no difference between the hMG and Grin groups. The single cyclophosphamide group presented loose tubules with pathologic vacuoles and significant TUNEL positive cells. Grin induced less weight of body or testis, compactly aligned tubules with little intra-lumens, whereas hMG caused more weight of body or testis, enlarging tubules with annular clearance. Grin presented a dose-dependent manner or cell differentiation-dependentincrease in testicular GHRH receptor, and did not impact the levels of blood and testicular GH, testosterone. Grin promotes fertility by proliferating and differentiating primitive cells through up-regulating testicular GHRH receptor without triggering GH secretion, which might solve the etiology of oligoasthenozoospermia.

Clinical Analysis of 40 Patients with Eosinophilic Lung Diseases in Peking Union Medical College Hospital / 中国医学科学院学报

Objective To summarize the clinical features of eosinophilic lung diseases(ELD). Methods We retrospectively analyzed the clinical manifestations,laboratory findings,accessory examination results,and pathology of 40 patients who were diagnosed with ELD and hospitalized in Peking Union Medical College Hospital from January 2013 to December 2016.Results There were 19 males and 21 females,and the average age was(48.58±18.25) years.The diagnoses included allergic bronchopulmonary aspergillosis(n=20),eosinophilic granulomatosis with polyangiitis(also known as churg-strauss syndrome)(n=10),chronic eosinophilic pneumonia(n=8),parasitic infection(n=1),and drug-induced eosinophilic pneumonia(n=1).Eosinophils counts in peripheral blood were increased in 35 patients(87.5%),and eosinophils counts in bronchoalveolar lavage fluids increased in 17 of 18 patients(94.4%).Arterial blood gas analysis showed varying degrees of hypoxemia in 23 patients(57.5%),and pulmonary function test showed ventilatory dysfunction in 27 patients(67.5%) and defect in diffusion capacity in 12 patients(30.0%).Chest CT revealed bilateral flaky,streaky,or diffuse ground-glass infiltrates and consolidations;in addition,central cylindrical bronchiectasis and mucous plugging with "finger-in-glove" pattern were seen in patients with allergic bronchopulmonary aspergillosis.Diffuse eosinophil infiltration was revealed in lung or other tissue biopsy.Glucocorticoids alone or combined with other therapies were effective in most patients.Conclusions ELD has a wide range of clinical presentations and can easily be misdiagnosed.Increased eosinophils count in peripheral blood and bronchoalveolar lavage fluids combined with infiltration manifestations in chest imaging are helpful for the diagnosis of ELD.Oral administration of glucocorticoids is the primary therapy for ELD.

Ferulic acid promoting apoptosis in human osteosarcoma cell lines

Objective: To explore the promoting apoptosis and antitumor activities of ferulic acid [FA] in human osteosarcoma and its potential mechanism

Methods: The SaOS-2 and MG63 osteosarcoma cell lines were opted to experiment and these cells were, respectively, cultured with various concentrations of FA [0 microM, 10 microM, 20 microM, 40 microM] for 72 hours at 37°C. The viabilities of the FA treated cells were monitored by MTT. Apoptosis cells were evaluated using annexin V/PI by flow cytometry. Apoptosis proteins caspase-3, procaspase-3, Bcl-2 and Bax were detected by western blot. Expressions of apoptotic genes Bcl-2 and Bax were quantified by qPCR

Results: The cell viabilities were critically declined in the concentration-dependent manner in FA groups [P < 0.01]. The apoptosis cells were increased proportionately with the concentration of FA [P < 0.05]. The procaspase-3 protein contents, and Bcl-2 mRNA and protein contents were significantly decreased while caspase-3 protein contents, and Bax mRNA and protein contents were concomitantly increased in the concentration-dependent manner in FA groups [P < 0.05]. The response to FA by the SaOS-2 osteosarcoma cell was similar with the MG63 osteosarcoma cell [P > 0.05]

Conclusion: Ferulic acid could significantly descend osteosarcoma cell viability through the promoting apoptosis pathway in which FA activates both caspase-3 and Bax and inactivates Bcl-2

Effects of siRNA-mediated HIF-1alpha gene silencing on angiogenesis in osteosarcoma

Objective: To explore angiogenesis in osteosarcoma under the condition of hypoxia-inducible factor [HIF]-1alpha gene silenced by small interference RNA [siRNA]

Methods: The SaOS-2 osteosarcoma cells, transfected with the recombinant plasmid p Silencer 2.1-HIF-1alpha or p Silencer 2.1-SCR, were classified as HIF-1alpha/siRNA group or SCR/siRNA group, respectively. In which, vascular endothelial growth factor [VEGF] immunohistochemistry were performed. HIF-1alpha and VEGF protein contents were detected by western blot. Gene expressions of HIF-1alpha and VEGF were quantified by qPCR. Then the transfected SaOS-2 cells were inoculated in nude mice and transplantation tumor were checked via HE staining, VEGF and CD34 immunohistochemistry, and calculation of microvascular density [MVD]

Results: In vitro, VEGF immunohistochemistry stains, HIF-1alpha and VEGF protein contents, and the relative expressions of HIF-1alpha mRNA and VEGF mRNA in HIF-1alpha/siRNA group were obviously reduced. In vivo, morphological observation illustrated that heteromorphism were not obvious in the cells of HIF-1alpha/siRNA group and vascular systems were sparse in its transplantation tumor tissue, and immunohistochemistry revealed that both VEGF and CD34 stains were significantly decreased in HIF-1alpha/siRNA group, and MVD in HIF-1alpha/siRNA group [7.3+/-1.1] were obviously less than that in SCR/siRNA group [17.2+/-3.2] [P<0.05]

Conclusion: Angiogenesis in osteosarcoma can be inhibited by siRNA-mediated HIF-1alpha gene silencing, which is expected to provide a novel and attractive target of therapeutic strategies of osteosarcoma