Objective To investigate the effect of high-mobility group box protein1 (HMGB1) on the expression of TNF-αand its mechanism in 16HBE in vitro. Methods groups with different HMGB1 (0, 100, 500, 2 000 ng/mL) concentration was set; RAGE antagonizing groups were as control, HMGB1-2000ng, anti-RAGE and anti-RAGE+HMGB1. The changes of TNF-αmRNA and secretion were determined by quantitative PCR and ELISA. RAGE protein level was measured by western blotting. Results HMGB1 intervention and TNF-α expression of 16HBE presented a positive dose-dependent relationship. Thechanges of RAGE was HMGB1positively concentration dependent. In comparison with HMGB1 2 000 ng/mL group, anti-RAGE+HMGB1showed a remarkable reduction of TNF-α secretion. Conclusion In vitro, HMGB1 increases TNF-α expression in 16HBE with a dose-dependent manner through RAGE.

BACKGROUND: Recent studies have proven the existence of the regeneration of central nervous tissue. But abjective results,such as lacked of neurogenesis after injury,also have been found in many experiments. The greatest difficulty in conventional brain transplantation or brain tissue transplantation has been the survival and development of the graft. Additionally,the stability of therapeutic effects and the rehabilitation of brain functions also need confirmation.OBJECTIVE: To investigate an approach to intrathecal injection of neural stem cells(NSCs) in stroke therapy,and observe therapeutic effects and side effects as well,so as to make the evaluation of the safety and feasibility.DESIGN: A confirmative before-after study based on stroke patients.SETTING: A neurology department in a municipal hospital and a microbiology and immunology department affiliated to a university hospital.PARTICIPANTS: From November 2002 to September 2003,26 stroke inpatients in the Neurology Department of Anyang Municipal People' s Hospital were selected. Of all the c ases,3 were diagnosed as acute cerebral hemorrhage,and the other 23 had been suffering strokes for durations ranging from 3months to 30 years,an average of (4.2 ± 6. 6) years. They were 20 male and 6female between the ages from 36 -72 years old,an average of(56.3 ± 12.7)years old. Fifteen of them were ischemic and 11 were hemorrhagic. Nineteen were associated with hypertension,2 with coronary heart disease,4 with diabetes and 4 associated with hyperlipodemia.INTERVENTIONS: On each of the 3 patients with acute cerebral hemorrhage (hematoma volume,35 - 40 mL),a microinvasive intracerebral hematoma puncture was performed,and then a suspension of NSCs were conducted to the stroke by a drainage tube. For the rest of the patients,suspensions were intrathecally administered into the subarachnoid and then flowed to the cerebral surface through cerebrospinal fluid(CSF) circulation. Afterwards,physical therapy(PT),occupational therapy (OT) and speech therapy(ST) were jointly applied to facilitate the rehabilitation of the stroke patients. Therapeutic effects was calculated according to the European stroke scale(ESS) and the Barthel Index(BI) . If ESS index went beyond or equal to 1 score,the case would be defined as effective; otherwise,it would be defined as not effective. Additionally,CT,MRI,EEG,chest x-ray,and blood biochemical variables were also measured.MAIN OUTCOMEMEASURES: Therapeutic effects and side effects were taken as main outcomemeasurements.RESULTS: Of the 23 patients who ntrathecal administration,19had positive therapeutic effect and 4 did not. Post-transplantation ESS was higher than that of pre-transplantation(54. 1 ±21.2 vs 51.4 ±21.1,t = 5.8,P = 0. 000 007 6),while post-transplantation BI also increased significantly as compared with that of pre-transplantation(41.1 ± 31.3 vs 36. 1 ± 32. 1,vasive intracerebral hematoma puncture had successful rehabilitation and regained self-care ability. Of all the patients,4 got a transitory fever and 2felt slight post-operation headache.CONCLUSION: Conclusion can be drawn from the study that stroke patients are ameliorated to various extents after neural stem cell transplantation which has no toxicity or side effects. It shows that neural stem cell transplantation is viable and feasible in improving the motor function and self-care ability in stroke patients.

In recent years, immunotherapy has made a breakthrough in the field of non-small cell lung cancer, reshaping the pattern of lung cancer treatment. However, with the wide application of immunotherapy in clinical practice, immune-related adverse events have attracted increasing attention. Immune pneumonia, as one of the immune-related toxic side effects of greatest concern, affects the treatment process and curative effect and can be a threat to life in serious cases. Given that immune pneumonia has a complicated pathogenesis and diverse clinical manifestations, strengthening the understanding of immune pneumonia is urgently needed. The treatment of immune pneumonia is limited, and additional therapeutic medicines are still awaiting exploration. Therefore, this paper summarizes the progress of the research on immune pneumonia in the treatment of non-small cell lung cancer.

ObjectiveTo establish a quality evaluation method for Zhuye Shigao granules（Zhuye Shigaotang） based on fingerprint and determination of index components， and to investigate the therapeutic effect of Zhuye Shigao granules on mice with cold-dampness pestilence attacking lung syndrome. MethodsThe fingerprint of Zhuye Shigao granules was established by high performance liquid chromatography（HPLC）， and the methods for determination of total calcium， orientin， isoorientin， ginsenosides Rg₁， Re and Rb₁ and other 2 index components were established. Fifty ICR mice were randomly divided into the blank group， model group， Zhuye Shigao granules low， medium and high dose groups（9.3， 18.6， 37.2 g·kg^-1·d^-1）， with 10 mice in each group. In addition to the blank group， the model mice with cold-dampness pestilence attacking lung syndrome was prepared by nasal drip of lipopolysaccharide combined with cold-dampness environment. Each administration group was given the corresponding liquid by gavage according to the dose， while the blank group and model group were given the same volume of normal saline by gavage. Then， the body temperature and organ index of mice in each group were measured， hematoxylin-eosin（HE） staining was used to investigate the lung tissue injury of mice in each group， and enzyme-linked immunosorbent assay（ELISA） was used to detect the changes of tumor necrosis factor-α（TNF-α）， interleukin（IL）-lβ， IL-6， IL-10 levels in serum and lung tissue， as well as immunoglobulin（Ig）A and IgM levels in serum. ResultsThe fingerprint similarity of 10 batches of Zhuye Shigao granules was>0.950， and 20 common peaks were calibrated. Seven of them were identified， including peak 11（isoorientin）， peak 12（orientin）， peak 14（apioside liquiritin）， peak 15（liquiritin）， peak 17（apioside isoliquiritin）， peak 19（isoliquiritin） and peak 20（liquiritigenin）. The results of quantitative analysis showed that the content range of each index component in 10 batches of Zhuye Shigao granules was as follows：Total calcium of 9.978-11.294 mg·g^-1， isoorientin of 0.033-0.041 mg·g^-1， orientin of 0.046-0.055 mg·g^-1， ginsenoside Rg₁+ginsenoside Re of 0.748-0.762 mg·g^-1， ginsenoside Rb₁ of 0.151-0.197 mg·g^-1， liquiritin of 1.106-1.366 mg·g^-1， glycyrrhizic acid of 0.904-1.182 mg·g^-1_. Compared with the blank group， the body temperature of mice in the model group was significantly increased， the organ indexes of liver， lung and spleen were significantly decreased， the organ index of thymus was significantly increased， HE staining of lung tissue showed infiltration of inflammatory cells， a small amount of serous exudation was observed in the alveoli， and lung tissue was damaged. After the intervention of Zhuye Shigao granules， the pathological changes were improved compared with the model group. The expression levels of IL-1β， IL-6 and TNF-α were significantly increased， the expression level of IL-10 was significantly decreased in serum and lung tissue. The levels of IgA and IgM in serum were significantly decreased（P<0.01）. Compared with the model group， the body temperature， the organ indexes and immune factor levels in serum and lung tissue of mice in the Zhuye Shigao granules medium and high dose groups were significantly reduced（P<0.05， P<0.01）. ConclusionIn this study， the quality evaluation of Zhuye Shigao granules was carried out based on fingerprint combined with determination of index components， and the fingerprint of four herbs（Lophatheri Herba， Ophiopogonis Radix， Pinelliae Rhizoma and Glycyrrhizae Radix et Rhizoma） in this formula and the determination of 8 index components were established. The therapeutic effect of Zhuye Shigao granules on mice with cold-dampness pestilence attacking lung syndrome may be related to inhibiting inflammatory response and mediating immune regulation.

To evaluate the prevalence of mosquito-borne viruses in Manshi and Ruili (Yunnan Province, China), we collected 2 149 mosquitoes (17 species) in August 2010. Virus isolation was undertaken by the cul- ture of baby hamster kidney cells (BHK-21 cells). Two virus-like isolates were obtained: DHL10M117 was isolated from collected in Mangshi; DHL10M110 was obtained from Anopheles vagus collected in Rui- li. Both isolates caused cytopathic effects,illness and death in suckling mice inoculated with these isolates via the intracerebral route. Two positive amplicons, 702-bp from the S segment and 456-bp from the M segment,were obtained using reverse transcription-polymerase chain reaction using primers specific for the Akabane virus (AKV). Phylogenetic analysis suggested that these two virus stains had a distant relation- ship with AKVs from Kenya and Australia,but were genetically close to those from Japan,South Korea, and Taiwan. However,they were separate from other Asian strains and grouped into a small branch. The highest nucleotide and amino-acid sequence identity of the S segment was found with the CY-77 strain from Taiwan (96.6% and 99.6% for DHL10M117 and 96.7% and 100% for DHL10M110,respectively). Com- parison of the M segment showed they shared the highest amino acid identity with CY-77 (99.6% and 100%, respectively), whereas the highest nucleotide identity was found with the Iriki strain from Japan (99.6% and 100%, respectively). Compared with the MP496 strain from Kenya,they displayed lower lev- els of sequence homology, at 69.7% and 70.0% for nucleotide sequences of the two loci,and 91. 0% for a- mino acids. Our results identified that DHL10M117 and DHL10M110 were strains of AKV,and provided molecular biological evidence for the existence of AKV in Yunnan Province. These AKV strains that are circulating in Yunnan Province share a close genetic relationship with strains from the rest of Asia. Culex tritaeniorhynchus and Anopheles vagus may serve as transmission vectors.
Amino Acid Sequence ; Animals ; Anopheles ; virology ; Base Sequence ; Bunyaviridae Infections ; virology ; China ; Cricetinae ; Female ; Humans ; Insect Vectors ; virology ; Male ; Mice ; Orthobunyavirus ; classification ; genetics ; isolation & purification ; physiology ; Phylogeny ; Sequence Homology ; Viral Proteins ; chemistry ; genetics

Injectable sustained-release pcDNA3-GRF (1-32) microspheres were prepared by double emulsion-in liquid evaporation process,using biodegrable poly lactic-co-glycolic acid as carrier. The enrapment efficiency, mean particle size, drug content thus prepared were 69%, 2.20 microm, 8% and 70% respectively. The result of transfection in vivo showed that after 30 days, accumulative increased body weights on the group injected with pcDNA3-GRF (1-32) microspheres was significantly higher than those group injected with naked plasmid (12.87%), plasmid-empty microspheres (19.72%) and saline (58.58%) respectively. PCR and RT-PCR showed that the expression level of GRF gene on the group injected with pcDNA3-GRF (1-32) microspheres was the highest. GRF gene released by microspheres was still detected after 30 days. In conclusion, pcDNA3-GRF (1-32) microspheres have a controlled release effect and GRF gene could be successfully transfected into muscle cells of mouse by microspheres with higher efficacy and stronger biological function.
Animals ; Body Weight ; DNA ; administration & dosage ; Growth ; Growth Hormone-Releasing Hormone ; genetics ; Lactic Acid ; administration & dosage ; Male ; Mice ; Microspheres ; Muscle, Skeletal ; metabolism ; Plasmids ; Polyglycolic Acid ; administration & dosage ; Polymerase Chain Reaction

Bone morphogenetic proteins(BMPs)belongs to the transforming growth factor beta(TGF-β)superfamily, which was isolated from the decalcified bone matrix by Urist in 1965.As one of the members of BMPs family, BMP4 not only promotes the differentiation and development of chondrocytes, but also plays an important role in the regulation of eye cells. BMP4 can mediate apoptosis of retinal vascular endothelial cells and inhibit angiogenesis, and is expressed in adult corneal epithelium, corneal cells and corneal endothelial cells. Also, participated in the layered corneal epithelium cells proliferation. In this paper, the recent advances in the study of BMP4 in ocular tissues are summarized, and the function of BMP4 in different developmental stages of eye is briefly reviewed.

ObjectiveTo investigate the pharmacokinetic characteristics of sofosbuvir tablets, and to evaluate the bioequivalence and safety of two preparations. MethodsHealthy volunteers were recruited through the platform of clinical trial recruitment in The Affiliated Hospital of Changchun University of Chinese Medicine. Screening physical examination was performed for fasting group on September 18, 2018 and for postprandial group on September 28, 2018, and the volunteers were enrolled after their physical examination results met the inclusion criteria. The fasting group and the postprandial group, with 40 volunteers in each group, were given oral administration of the test preparation sofosbuvir tablets or the reference preparation sofosbuvir tablets (SOVALDI, 400 mg). This was a randomized, open-label, two-sequence, four-cycle, single-dose, and completely repeated cross-over bioequivalence test in the fasting or postprandial state in the healthy population; in the fasting group, 20 volunteers each received oral administration of the test preparation and the reference preparation, and in the postprandial group, 20 volunteers each received oral administration of the test preparation and the reference preparation. Liquid chromatography-tandem mass spectrometry was used to measure the content of sofosbuvir and its major metabolite GS-331007 in human EDTA-K2 plasma; the plasma concentration of sofosbuvir was measured at 15 time points from 0 hour to 8 hours after administration, and that of GS-331007 was measured at 16 time points from 0 hour to 72 hours after administration. WinNonlin software was used to calculate pharmacokinetic parameters and evaluate bioequivalence. ResultsAfter the administration of the test preparation and the reference preparation in the fasting state, when the pharmacokinetic parameters of sofosbuvir was used to evaluate the bioequivalence of the test preparation and the reference preparation, the ratios of the geometric means of Cmax, AUC0-t, and AUC0-inf were 90.55%, 97.26%, and 94.62%, respectively; when the pharmacokinetic parameters of GS-331007 was used to evaluate the bioequivalence of the test preparation and the reference preparation, the ratios of the geometric means of Cmax, AUC0-t, and AUC0-inf were 98.91%, 98.98%, and 99.46%, respectively. All of the above values were within the range of 80.00%-125.00%. An analysis of variance was performed after the pharmacokinetic parameters of sofosbuvir Cmax, AUC0-t, and AUC0-inf were transformed by natural logarithm, and the results showed that sequence, cycle, and preparation had no marked influence on Cmax, AUC0-t, and AUC0-inf (all P＞0.05). ConclusionThe test preparation of sofosbuvir tablets is bioequivalent to the reference preparation in the fasting and postprandial states.

OBJECTIVEThe aim of this study was to evaluate the effect of postoperative adjuvant therapy on the survival in patients with N1 lymph node metastasis of esophageal squamous cell carcinoma (ESCC).

METHODS110 patients with positive N1 lymph node metastasis of esophageal squamous carcinoma were included in this study. The surgery group included 46 cases and the postoperative adjuvant therapy group included 64 cases (24 cases in the adjuvant chemotherapy subgroup and 40 cases in the adjuvant concurrent chemoradiotherapy). The disease-free survival (DFS) and overall survival (OS) of the two groups were compared and the prognostic factors were analyzed by multivariate Cox model.

RESULTSIn the postoperative adjuvant therapy group, the DFS (16.8 months) and OS (21.3 months) were significantly prolonged compared with those in the surgery group (10.6 months, P=0.007) and (13.7 months, P=0.001), respectively. Postoperative adjuvant chemotherapy significantly extended the OS (31.1 months) of N1-positive patients compared with 13.7 months (P=0.002) in the surgery group. But there were no significant differences between the DFS in the two subgroups (16.3 and 16.8 months, P=0.346) and between the OS (23.4 and 21.3 months, P=0.491). Postoperative adjuvant therapy was an independent prognostic factor in the ESCC patients with N1 lymph node metastasis.

CONCLUSIONPostoperative adjuvant therapy can improve the prognosis and prolong the survival time in ESCC patients with positive N1 lymph node metastasis.

Carcinoma, Squamous Cell ; mortality ; secondary ; therapy ; Chemoradiotherapy ; Chemotherapy, Adjuvant ; Disease-Free Survival ; Esophageal Neoplasms ; mortality ; pathology ; therapy ; Humans ; Lymph Nodes ; Lymphatic Metastasis ; Postoperative Care ; Prognosis ; Retrospective Studies

Humans ; Joint Dislocations ; Lunate Bone ; Wrist Injuries