Research progress on stereotactic radiotherapy combined with immunotherapy for malignancies / 中华放射肿瘤学杂志

Surgery,radiotherapy and chemotherapy are three traditional treatments for malignant tumors.With the development of medicine,immunotherapy has been gradually adopted as an emerging therapy of malignancies.Recent clinical studies have demonstrated that the combination of radiotherapy and immunotherapy can induce the abscopal effect and improve the prognosis of patients.Compared with the conventional radiotherapy,stereotactic radiotherapy has a larger single dose and higher accuracy,which is more likely to induce the bystander effect and anti-tumor response.The combination of stereotactic radiotherapy and immunotherapy has been proven to be a more promising therapy in certain clinical trials.However,not all types of tumors can benefit from such combined therapy in clinical practice.The optimal dose,fraction pattern and lesion of radiotherapy,immune enhancement and safety remain to be further clarified.In this article,the research progress,related controversies and future research direction of stereotactic radiotherapy combined with immunotherapy for malignancies were reviewed.

Amplification and identification of regulatory T cells derived from mice in vitro / 中国医师杂志

Objective@#To establish a method for in vitro expansion of regulatory T cells (Tregs) for clinical study and immunotherapy.@*Methods@#CD4+ T cells were isolated from BALB/c spleens by magnetic activated cell sorting (MACS), then cultured in 24-well plates coated with anti-CD3/CD28 microbeads in the presence of 100 U/ml interleukin-2 (IL-2) and 5 ng/ml recombinant human transforming growth factor-β (rhTGF-β). The purity of the expanded Tregs were tested by flow cytometry. In vitro inhibition of CD4+ CD25-T cells response by expanded Tregs was measured by mixed lymphocyte reaction test. The number and the viability of the expanded Tregs were detemined by trypan blue staining.@*Results@#Tregs were expanded up to 20 folds after 5 days in culture, and the activity was (93±4)%. The purity of expanded Tregs were (79.1±1.5)%, and maintained suppressive ability.@*Conclusions@#We can obtain large numbers of Tregs with highly purity and suppressive ability, thereby providing a solution to the availability of sufficient Tregs in clinical study and immunotherapy.

Long-term disease-free survival in patient with laryngeal small cell neuroendocrine carcinoma: A case report and literature review / 吉林大学学报(医学版)

Objective: To analyze the clinical characteristics of patient with laryngeal small cell neuroendocrine carcinoma (LSCNC) with long-term disease-free survival, and to provide the basis for the diagnosis and treatment of LSCNC. Methods: The clinical materials of a patient with primary LSCNC were collected, and the clinical features, diagnosis and treatment of LSCNC and extrapulmonary small cell cancer (EPSCC) combined with the relative literatures were analyzed. Results: The patient was a 55-year-old man who had a long history of heavy smoking and drinking, and presented with neck mass. The laryngoscope revealed the uneven mass on the left side of epiglottic laryngeal surface. The subsequent biopsy of the neck mass indicated lymph node metastasis. The patient underwent laryngectomy and bilateral cervical lymph node dissection. The postoperative pathology indicated LSCNC; the immunohistochemistry results showed CD56 (NK-1) (+), CK-pan (+), CgA (-), Syn (-) and Ki-67 (+ 90%). The patient underwent radiotherapy to the tumor bed and lymph node drainage area and 6 courses of chemotherapy with EP regimen. No prophylactic cranial irradiation (PCI) was conducted. Regular follow-up with laryngoscope, crainial MRI and other examinations showed no recurrence and metastasis. The patient still alive with disease-free survival of 38 months now. Conclusion: LSCNC is highly malignant. The diagnosis mainly depends on the pathohistomorphology and immunohistochemistry examination. The combination of surgery, chemotherapy and radiotherapy is commonly used for LSCNC at present. PCI is not recommended routinely. Multimodality treatment can improve the prognosis.

Amplification and identification of regulatory T cells derived from mice in vitro / 中国医师杂志

Objective To establish a method for in vitro expansion of regulatory T cells (Tregs)for clinical study and immunotherapy.Methods CD4 + T cells were isolated from BALB/c spleens by magnetic activated cell sorting (MACS),then cultured in 24-well plates coated with anti-CD3/CD28 microbeads in the presence of 100 U/ml interleukin-2 (IL-2) and 5 ng/ml recombinant human transforming growth factor-β (rhTGF-β).The purity of the expanded Tregs were tested by flow cytometry.In vitro inhibition of CD4 + CD25-T cells response by expanded Tregs was measured by mixed lymphocyte reaction test.The number and the viability of the expanded Tregs were detemined by trypan blue staining.Results Tregs were expanded up to 20 folds after 5 days in culture,and the activity was (93 ± 4) ％.The purity of expanded Tregs were (79.1 ± 1.5) ％,and maintained suppressive ability.Conclusions We can obtain large numbers of Tregs with highly purity and suppressive ability,thereby providing a solution to the availability of sufficient Tregs in clinical study and immunotherapy.

Erratum to: The crystal structure of Ac-AChBP in complex with α-conotoxin LvIA reveals the mechanism of its selectivity towards different nAChR subtypes

In the original publication of the article the keywords are incorrectly online published. The correct keywords should read as α-Conotoxin; Nicotinc acetylcholine receptor; Acetylcholine binding protein; X-ray crystallography".

The crystal structure of Ac-AChBP in complex with α-conotoxin LvIA reveals the mechanism of its selectivity towards different nAChR subtypes

The α3* nAChRs, which are considered to be promising drug targets for problems such as pain, addiction, cardiovascular function, cognitive disorders etc., are found throughout the central and peripheral nervous system. The α-conotoxin (α-CTx) LvIA has been identified as the most selective inhibitor of α3β2 nAChRs known to date, and it can distinguish the α3β2 nAChR subtype from the α6/α3β2β3 and α3β4 nAChR subtypes. However, the mechanism of its selectivity towards α3β2, α6/α3β2β3, and α3β4 nAChRs remains elusive. Here we report the co-crystal structure of LvIA in complex with Aplysia californica acetylcholine binding protein (Ac-AChBP) at a resolution of 3.4 Å. Based on the structure of this complex, together with homology modeling based on other nAChR subtypes and binding affinity assays, we conclude that Asp-11 of LvIA plays an important role in the selectivity of LvIA towards α3β2 and α3/α6β2β3 nAChRs by making a salt bridge with Lys-155 of the rat α3 subunit. Asn-9 lies within a hydrophobic pocket that is formed by Met-36, Thr-59, and Phe-119 of the rat β2 subunit in the α3β2 nAChR model, revealing the reason for its more potent selectivity towards the α3β2 nAChR subtype. These results provide molecular insights that can be used to design ligands that selectively target α3β2 nAChRs, with significant implications for the design of new therapeutic α-CTxs.

Activities of α-conotoxin TxID isomers on human nicotinic acetylcholine receptors / 中国药科大学学报

@#To investigate activities of three isomers of α-conotoxin TxID on human α3β4 and α6/α3β4 nicotinic acetylcholine receptors(nAChRs). The three isomers of α-conotoxin TxID were synthesized using solid phase Fmoc chemistry and fully folded by two-step oxidations. Human α3β4 and α6/α3β4 nAChRs were expressed in oocytes of Xenopus laevis, which were used for bioassay of the three isomers, including inhibition and washout reversibility. There were obvious differences between the inhibition potency of each isomers at human α3β4 and α6/α3β4 nAChRs. The blocking was reversible and washout rapidly. The most potent isomer is the globular form with an IC50 of 9. 3 nmol/L on human α3β4 and α6/α3β4 nAChRs respectively. The 2nd potent isomer was the ribbon form with much less potency, which had an IC50 of > 5 μmol/L. The bead isomer had little or no block on human α3β4 and α6/α3β4 nAChRs with an IC50 of > 10 μmol/L. The three isomers of α-conotoxin TxID were synthesized successfully with two pairs of desired disulfide bond. Inhibition activities of the 3 isomers on human α3β4 and α6/α3β4 nAChRs were obtained respectively, which would be basis for new marine drug development of α-conotoxin TxID.

Synthesis and evaluation of a novel injectable and water-swelling gingival displacement materials / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To synthesize and evaluate a novel injectable and water-swelling gingival displacement materials.</p><p><b>METHODS</b>A kind of water-swelling polymer, kaolin and aluminum chloride were mechanically mixed at certain ratio in water solution, resulting to a novel paste materials for gingival displacement. Then, its stability in aqueous solution and water swelling properties were evaluated in vitro. The effect on gingival displacement was evaluated by animal experiments in dogs. A commercial gingival displacement materials paste of Expasyl was used as control.</p><p><b>RESULTS</b>While contacting with water, the novel gingival displacement paste did not collapse, maintained its integrity structure, and could expand for adsorbing water. Animal experiments in dogs showed that the materials could lead to displace the gingival margins from the dental root surfaces.</p><p><b>CONCLUSION</b>The novel injectable and expanded gingival displacement material is efficient to retract free gingival margin with potential clinical application.</p>

Removing action of compound danshen di wan on carotid atherosclerotic plaques: Multi-central randomized controlled experiment / 中国组织工程研究

BACKGROUND: It is indicated in a large amount of basic researches that compound danshen di wan has a certain action on removing carotid atherosclerotic plaques (CAP), but there is lack of large sample data in randomized, controlled, multi-central clinical experiment.OBJECTIVE: To observe the removing action of compound danshen di wan on CAP, compared with aspirin.DESIGN: Multi-central, randomized controlled experiment was designed.SETTING: Research Center of Traditional Chinese Medicine and Ultrasonic Department of Xinjing Hospital, Fourth Military Medical University of Chinese PLA; Department of Traditional Chinese Medicine of Second Clinical Hospital, Xi' an Jiaotong University and Department of Cardiac Internal Medicine of Xi' an Hospital of Traditional Chinese Medicine.PARTICIPANTS: Totally 162 cases were selected from the clinic of Research Center of Traditional Chinese Medicine, Fourth Military Medical University of Chinese PLA; Department of Traditional Chinese Medicine, Second Clinical Hospital, Xi' an Jiaotong University and Department of Cardiac Internal Medicine of Xi' an Hospital of Traditional Chinese Medicine from 2002 to 2004, aged varied from 40 to 80 years, the thickness of carotid internal media ≥ 1.2 mm. They were randomized into two groups, named danshen di wan group of 89 cases(cases of soft plaque and hard plaque were 49 and 40 respectively) and aspirin group of 73 cases(cases of soft plaque and hard plaque were 42 and 31 respectively).METHODS: In danshen di wan group: compound danshen di wan was administrated orally(10 pills/time, 3 times/day), in aspirin group, enteric soluble aspirin table was administrated orally(75 mg/time, once a day),continuously for 6 months. The alternations of thickness of carotid internal media were determined with ultrasonic B of high resolving power before and after treatment.MAIN OUTCOME MEASURES: Changes in thickness of carotid internal media before and after treatment.RESULTS: By practical measuring analysis, 143 cases accomplished nation of thickness of carotid internal media in the patients with soft plaque:In danshen di wan group, the thickness was remarkably decreased after treatment[ (2. 12 ± 0. 34), (2.44± 0.40) mm, t = 4. 267, P ＜ 0.01 ] . In aspirin group, it was also reduced relatively after treatment[ (2.25 ± 0. 3),of carotid internal media in the patients with hard plaque: The changes were not significant no matter in danshen di wan group or aspirin group in the comparison before and after treatment[in danshen di wan group: (2.46 ±0.42),(2.34 ± 0. 40) mm; in aspirin group: (2.42 ± 0. 44), (2. 36 ± 0. 38) mm,P＞ 0.05].CONCLUSION: Compound danshen di wan acts on removing soft atherosclerotic plaque and its effect is similar to aspirin.

Cloning of novel O-superfamily conotoxin of Conus capitaneus collected from Hainan / 中国海洋药物

Objective To isolate and clone novel O-superfamily conotoxin genes of Conus capitaneus Linnaeus collected from Hainan,which would provide the initial drug leads to investigate and develop Conus capitaneus conotoxins.Methods 3'-RACE(Rapid Amplification of cDNA Ends) was used for cloning the novel O-superfamily conotoxins.The specific amplified cDNA fragments were sequenced and analyzed,as well as the genetic diversity of the O-superfamily conotoxins.Results The full-length cDNA(CaHr91N) of a new O-superfamily conotoxin(CaHr91) was cloned and sequenced from Conus capitaneus Linnaeus.The novel conotoxin precursor CaHr91P with 77 amino acids(aa) encoded by the cDNA consists of three typical regions of signal with 21aa, pro-peptide with 22aa and mature peptide with 34aa.Predicted sequence of the toxin region CaHr91M is "ECREQSQGC TNTSPPCC SGLRC SGQSQGGVC ISN" with a common O-superfamily cysteine pattern C-C-CC-C-C.Percent identities between CaHr91P and other published homologue O-superfamily sequences were compared systematically,as well as research status on conopeptides from C.capitaneus.Conlusion The elucidated cDNA of the novel CaHr91P conotoxin will be the basis for a better understanding of its bioactivity and application,as well as finding more novel conotoxins from C.capitaneus.