1.Construction and identification of recombinant adenovirus containing human vascular endothelial growth factor 165 gene
Yanfu HAN ; Jianxing SONG ; Jun LIU
Chinese Journal of Tissue Engineering Research 2007;0(24):-
AIM: Recombinant adenovirus possesses high transfection efficiency and wide host range. This study was designed to construct the recombinant adenovirus vector containing human vascular endothelial growth factor 165 (VEGF165), so as to lay a foundation for the subsequent gene transfection, microencapsulated genetically engineered cells and animal experiments. METHODS: The experiment was conducted in the Laboratory of Cardiothoracic Surgery (the National Key Laboratory), Changhai Hospital of The Second Military Medical University of Chinese PLA from January to May in 2007. Experiment materials: pAxCAwt.VEGF165 was provided by Institute of Cardiothoracic Surgery of Changhai Hospital. pAxCAwt.VEGF165 and DNA-TPC were cotransfected into human embryonic kidney 293 cells by lipofection method. Being propagated, recombinant replication-deficient adenovirus named Ad.VEGF165 was obtained. The target gene of recombinant adenovirus was identified by polymerase chain reaction (PCR) and restriction enzyme digestion. The titer of virus was detected by 50% tissue culture infective dose method. RESULTS: Construction of recombinant adenovirus Ad.VEGF165: The pAxCAwt.VEGF165 and DNA-TPC were successfully cotransfected into human embryonic kidney 293 cells by lipofection method, and replication-deficient adenovirus vectors coding for VEGF165 gene were generated. Identification of recombinant adenovirus Ad.VEGF165: Two fragments of PCR products (597 bp and 146 bp) were obtained by NcoI restriction enzyme. The result was consistent with that calculated with Gene Tool software. The virus titers was 2.2?1015 pfu/L. CONCLUSION: DNA-TPC and pAxCAwt.VEGF165 can be used to construct replication-deficient recombinant adenovirus Ad.VEGF165 in a high titer, low toxicity, high efficiency and safe transfection in vitro.
2.Effects of extract from fermented buckwheat flower and leaf(EFBFL)on myocardial injury in type 2 diabetic db/db mice
Jianxing WANG ; Yan WANG ; Man LIU ; Yu MU ; Bo ZHANG ; Shuying HAN
Chinese Pharmacological Bulletin 2017;33(7):1026-1031
Aim To discuss the effectsof extract from fermented buckwheat flower and leaf(EFBFL) on myocardial injury in spontaneously obese type Ⅱ diabetic db/db mice and its mechanism.Methods 9-week-old male db/db mice were randomly divided into high level EFBFL dose group(EFBFL-H, 0.1 g·kg-1), low level EFBFL dose group(EFBFL-L, 0.05 g·kg-1),metformin hydrochloridecontrol group, model control group, and normal control group, with 10 mice in each group.All groups were treated with 8 wks of drugs by gastric perfusion.The random blood glucose(RBG) was tested respectively at the end of 2nd, 4th, 6th, and 8th wk.Finally, the levels of creatine kinase(CK) creatine kinase MB(CK-MB), andadvanced glycosylation endproducts(AGEs) were detected after 8 wks.The morphological changes of myocardium were observed under light microscope by HE staining, and the ultrastructure of myocardium was observed under electron microscope.Immunohistochemical method and Western blot were used to detect myocardial tissue glucose transporter-4(Glut-4).Results EFBFLcould repress patho-proceeding of myocardial fibrosis efficiently, and significantly decrease the level of blood glucose, CK,CK-MB, and AGEs in db/db mice.Meanwhile, it could increase the expression of Glut-4 in myocardial tissues of mice.Conclusions EFBFL can prevent myocardial injury in spontaneously obese type Ⅱ diabetic db/db mice.The possible mechanism may be related to lowering the level of blood glucose and serum AGEs and up-regulating Glut4of cardiac muscle.
3.Gene mutations and prenatal diagnosis in families with maple syrup urine disease
Nan YANG ; Liqin ZHANG ; Lianshu HAN ; Jun YE ; Wenjuan QIU ; Huiwen ZHANG ; Zhuwen GONG ; Yafen ZHANG ; Jianxing ZHU ; Xuefan GU
Chinese Journal of Perinatal Medicine 2012;15(8):494-499
Objective Maple syrup urine disease (MSUD) is a rare metabolic disorder caused by deficiency of the activity of branched-chain 2-keto acid dehydrogenase complex.The complex contains E1α,E1β and E2 subunits which are encoded by BCKDHA,BCKDHB or DBT genes respectively.Mutation in any gene will cause MSUD.The aim of this study was to analyze the gene mutations of four cases with MSUD and carry out prenatal diagnosis for these four families for MSUD.Methods From 2005 to 2010,four neonates (two males and two females) were diagnosed as MSUD at 2,5,10and 26 days of life.The coding regions of BCKDHA gene and BCKDHB gene in the above four cases were amplified by polymerase chain reaction and analyzed by direct DNA sequencing.During the second pregnancy of the same mother,the amniotic fluid was drawn out at 16-20 weeks for gene mutation analysis after the amniocytes were cultured.Results Mutation analysis revealed six mutations in four patients,including four novel mutations (c.308T>C,c.562G>T,c.1279C>G and c.1280-1291de112) and two previously reported mutations.Five mutations (c.308T>C,c.562G >T,c.868G>A,c.1279C>G and c.1280-1291de112) were detected on BCKDHA gene in three patients.While one mutation (c.853C>T) was found on BCKDHB gene in one patient.Only one mutation was found in the amniocytes of each patient's mother at their second pregnancies suggesting a MSUD heterozygous fetus.Conclusions Analysis of BCKDHA and BCKDHB allowed preliminary understand of gene mutations in the four MSUD families,and made prenatal diagnosis possible,which helped in consultation in the second pregnancy.
4.Therapeutic effects of oral appliance combined with uvulopalatopharyngoplasty on obstructive sleep apnea hypopnea syndrome.
Jianxing HAN ; Bei WANG ; Yunfang AN ; Yuan MA ; Yan LIU ; Yufeng MA
Chinese Journal of Stomatology 2014;49(2):69-72
OBJECTIVETo explore the long-term effects of the uvulopalatopharyngoplasty (UPPP) combined with oral appliance (OA) in the treatment of obstructive sleep apnea hypopnea syndrome (OSAHS).
METHODSThirty patients with severe OSAHS confirmed by polysomnography (PSG) according to their apnea hypopnea index (AHI) and lowest SaO2 during sleep were selected using random permutation table and divided into only UPPP group(U group, n = 15) and UPPP with OA group (UA group, n = 15). The follow-up period was 2 years. PSG was performed in 0.5 year and 2 years after operation. AHI, lowest SaO2 and Tmax were tested and evaluated.
RESULTSThe effectiveness of two groups was the same after 0.5 year. Two years after operation, the values [AHI: (18.06 ± 2.24) times/h; lowest SaO2: (88.64 ± 10.37)% and Tmax: (20.5 ± 17.6) s] in UA group were better than that [AHI: (49.73 ± 3.35) times/h; lowest SaO2: (79.56 ± 4.87)% and Tmax: (41.3 ± 19.7) s] in U group. The number of effectiveness was 9 and the number of ineffectiveness was 6 in U group, while in UA group, the number of effectiveness was 14 and the number of ineffectiveness was 1(P < 0.05).
CONCLUSIONSLong-term result of combined treatment was better than that of UPPP only.
Humans ; Orthodontic Appliances ; Palate ; surgery ; Pharynx ; surgery ; Polysomnography ; Sleep ; Sleep Apnea, Obstructive ; surgery ; Uvula ; surgery
5.Transnasal endoscopic marsupialization for treatment of maxillary cysts.
Yongjin JI ; Qingfeng LI ; Jianxing HAN ; Changqing ZHAO
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2012;26(17):798-800
OBJECTIVE:
To evaluate the effectiveness and usefulness of transnasal endoscopic surgery for the treatment of maxillary cysts.
METHOD:
Transnasal endoscopic surgery was performed in 13 patients with maxillary cysts that extended to the maxillary sinus or the nasal bottom. Five patients had a radicular cyst, three patients had a dentigerous cyst, three patients had a nasolabial cyst and two patients had a median cyst. After the resection of anterior edge of the inferior turbinate or the nasal bottom, the lateral wall of the inferior nasal meatus was opened. Then, the cyst wall of the maxillary sinus was partially or completely removed under the endoscope.
RESULT:
The cyst walls were completely or partial removed in 13 patients with maxillary cysts. There were no complications, and postoperative courses were uneventful. The follow-up period ranged from 6 to 36 months, and no recurrence were noted in any of the cases.
CONCLUSION
Endoscopic transnasal surgery for the maxillary cyst is less invasive than conventional dental approach, and most of the affected teeth can be preserved. This technique appears to be a simple and highly effective surgical treatment for the treatment of patients with maxillary cysts that extend to the maxillary sinus or the nasal bottom.
Adolescent
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Adult
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Cysts
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surgery
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Endoscopy
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methods
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Female
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Humans
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Male
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Maxilla
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surgery
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Middle Aged
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Nasal Cavity
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surgery
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Young Adult
6. Effects of anteriolateral thigh perforator flap and fascia lata transplantation in combination with computed tomography angiography on repair of electrical burn wounds of head with skull exposure and necrosis
Xiaoqing LI ; Xin WANG ; Yalong HAN ; Gang JI ; Zonghua CHEN ; Jia ZHANG ; Jianping ZHU ; Jianxing DUAN ; Yongjing HE ; Xiaomin YANG ; Wenjun LIU
Chinese Journal of Burns 2018;34(5):283-287
Objective:
To explore the effects of anteriolateral thigh perforator flap and fascia lata transplantation in combination with computed tomography angiography (CTA) on repair of electrical burn wounds of head with skull exposure and necrosis.
Methods:
Seven patients with head electrical burns accompanied by skull exposure and necrosis were admitted to our burn center from March 2016 to December 2017. Head CTA was performed before the operation. The diameters of the facial artery and vein or the superficial temporal artery and vein were measured, and their locations were marked on the body surface. Preoperative CTA for flap donor sites in lower extremities were also performed to track the descending branch of the lateral circumflex femoral artery with the similar diameter as the recipient vessels on the head, and their locations were marked on the body surface. Routine wound debridement and skull drilling were performed successively. The size of the wounds after debridement ranged from 12 cm×8 cm to 20 cm×12 cm, and the areas of skull exposure ranged from 8 cm×6 cm to 15 cm×10 cm. Anteriolateral thigh perforator flaps with areas from 13 cm×9 cm to 21 cm×13 cm containing 5-10 cm long vascular pedicles were designed and dissected accordingly. The fascia lata under the flap with area from 5 cm×2 cm to 10 cm×3 cm was dissected according to the length of vascular pedicle. The fascia lata was transplanted to cover the exposed skull, and the anteriolateral thigh perforator flap was transplanted afterwards. The descending branch of the lateral circumflex femoral artery and its accompanying vein of the flap were anastomosed with superficial temporal artery and vein or facial artery and vein before the suture of flap. The flap donor sites were covered by intermediate split-thickness skin graft collected from contralateral thigh or abdomen.
Results:
The descending branch of the lateral circumflex femoral artery and its accompanying vein were anastomosed with superficial temporal artery and vein in six patients, while those with facial artery and vein in one patient. All the flaps survived after the operation, and no vascular crisis was observed. Wound healing was satisfactory. One patient was lost to follow up. Six patients were followed up for 6 to 10 months. The patients were bald in the head operation area with acceptable appearance. No psychiatric symptom such as headache or epileptic seizure was reported. The flap donor sites were normal in appearance. The muscle strength of the lower extremities all reached grade V. The sensation and movement of the lower extremities were normal.
Conclusions
Anterolateral thigh perforator flap with fascia lata transplantation can effectively repair electrical burn wounds of head with skull exposure and necrosis. The fascia lata can be used to protect the vascular pedicle of flaps, which is beneficial to the survival of the flap. Preoperative head and lower extremities CTA can provide reference for intraoperative vascular exploration in donor site and recipient area, so as to shorten operation time.
7.Anlotinib as third- or further-line therapy for short-term relapsed small-cell lung cancer: subgroup analysis of a randomized phase 2 study (ALTER1202).
Jianhua SHI ; Ying CHENG ; Qiming WANG ; Kai LI ; Lin WU ; Baohui HAN ; Gongyan CHEN ; Jianxing HE ; Jie WANG ; Haifeng QIN ; Xiaoling LI
Frontiers of Medicine 2022;16(5):766-772
Patients with small-cell lung cancer (SCLC) relapse within months after completing previous therapies. This study aimed to investigate the efficacy and safety of anlotinib as third- or further-line therapy in patients with short-term relapsed SCLC from ALTER1202. Patients with short-term relapsed SCLC (disease progression within 3 months after completing ⩾ two lines of chemotherapy) in the anlotinib (n = 67) and placebo (n = 34) groups were analyzed. The primary endpoint was progression-free survival (PFS). The secondary endpoints included overall survival, objective response rate (ORR), disease control rate, and safety. Anlotinib significantly improved median PFS/OS (4.0 vs. 0.7 months, P < 0.0001)/(7.3 vs. 4.4 months, P = 0.006) compared with placebo. The ORR was 4.5%/2.9% in the anlotinib/placebo group (P = 1.000). The DCR in the anlotinib group was higher than that in the placebo group (73.1% vs. 11.8%, P < 0.001). The most common adverse events (AEs) were hypertension (38.8%), loss of appetite (28.4%), and fatigue (22.4%) in the anlotinib group and gammaglutamyl transpeptidase elevation (20.6%) in the placebo group. No grade 5 AEs occurred. For patients with short-term relapsed SCLC, third- or further-line anlotinib treatment was associated with improved survival benefit. Further studies are warranted in this regard.
Humans
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Lung Neoplasms/drug therapy*
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Treatment Outcome
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Neoplasm Recurrence, Local/chemically induced*
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Quinolines/adverse effects*