Objective To construct recombinent retroviral vectors containsins the expression sequence of hIL-4,and detect the expression of target genes.Methods Design the primer with restrictive enzyme spot and amplify target gene by PCR from plasmid including hIL-4 gene.Clone the target gene into retroviral vector pLXSN,and identify the aquired plasmid by sequencing.Transfect human synoviocytes with acquired virus in vitro.Detect the protein expresssion by western blotting.Results We successfully constructed the recombinant retrovirus-rRV-hIL-4.The hIL-4 gene was transduced into human synoviocytes by recombinant retrovirus in vitro.The protein expression of genes was detected by Western blotting.Conclusion Recombinant retrovirus rRV-hIL-4 is constructed successfully.The hIL-4 gene is transduced successfully into the human synoviocytes in vitro and the transduced synoviocytes can express hIL-4 protein.

Objective To evaluate of the clinical efficacy of two kinds of methods extracorporeal shock wave lithotripsy in treatment of ureteral calculi. Methods A retrospective analysis was conducted on the data of 108 patients with acute renal colic and ureteral stones (calculi diameter<2 cm) from March 2011 to April 2013. The patients were divided into two groups. 69 cases in A group underwent emergency positioning urography ureteral catheterization combined extracorporeal shock wave lithotripsy, 39 cases in B group underwent emergency extracorporeal shock wave lithotripsy. Results A group:lithotripsy was successful in 63 cases among 69 cases at the first time,the first success rate was 91.33% (63/69), the second success rate was 94.2%(65/69),2 cases of retrograde ureteroscopy failed, 2 cases of self-discharge after intubation stones, the recurrent renal colic rate was 14.3%(9/63),the use of analgesic drugs rate was 11.1%(7/63) . B group:the first success rate of stone clearance rate was 74.4%(25/39),the second success rate of stone clearance rate was 87.2%(34/39),5 cases underwent ureteroscopy lithotripsy, the renal colic recurrence rate was 20.5% (8/39), the use rate of analgesic drugs was 15.4% (6/39) . Conclusion Extracorporeal shock wave lithotripsy assisted by urography through retrograde ureteral catheterization has satisfying therapeutic effect in treatment of patients with ureteral calculi.

BACKGROUND:Scaffolds made of chitosan and its derivatives play an important role in cellmigration and axonal regeneration. Chitosan and its derivatives have good histocompatibility, which is easy to make stem cells grow on the surface, thereby having a more broad application prospect in the nerve tissue engineering.
OBJECTIVE:To fabricate a thermosensitive hydrogel scaffold using chitosan/hydroxypropyltrimethyl ammonium chloride chitosan/sodium glycerophosphate (CS/HACC/GP), which is suitable for cellgrowth, and then, to observe the growth and proliferation of bone marrow mesenchymal stem cells on the scaffold.
METHODS:Chitosan was modified using quaternary ammonium salt and confirmed by Fourier transform infrared spectroscopy. The chitosan and quaternary ammonium salt of chitosan was mixed at a ratio of 8:1 to successful y prepare stable CS/HACC/GP thermosensitive hydrogel scaffold. Then, the gel ing was observed, and biosafety test was conducted.
RESULTS AND CONCLUSION:Fourier transform infrared spectroscopy showed the characteristic peak of quaternary ammonium groups. Cytotoxicity test showed that rat bone marrow mesenchymal stem cells cultured in hydrogel extracts had no toxicity. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test showed that hydrogel extracts exerted no significant effect on the increase in body weight, and the biological safety of the scaffold was good. Under the scanning electron microscopy, bone marrow-derived mesenchymal stem cells grew and proliferated normal y in the scaffold. The results confirmed that the CS/HACC/GP thermosensitive hydrogel scaffold was successful y prepared in the experiment, which is suitable for the growth and proliferation of bone marrow mesenchymal stem cells.

BACKGROUND:In recent years, chitosan-based thermosensitive hydrogel, as scaffold materials, have received more and more attentions in the field of tissue repair because of good biocompatibility, biodegradability and drug-sustained release.
OBJECTIVE:To explore the directed differentiation and growth of rat bone marrow mesenchymal stem cells on the quaternary chitosan thermosensitive hydrogel scaffold and to look for more ideal tissue engineering materials for the treatment of nervous system damage.
METHODS:The thermosensitive hygrogel scaffold was prepared using hydroxypropyltrimethyl ammonium chloride chitosan (HACC) andβ-glycerophosphate (β-GP). The spatial structure of scaffold was observed by scanning electronic microscope. Effect of leaching liquor from the HACC/β-GP scaffold on the viability of bone marrow mesenchymal stem cells was detected by (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The albumin from bovine serum was combined with the scaffold, and the slow-release effect of the scaffold was detected by ultraviolet absorption spectrometry. Bone marrow mesenchymal stem cells were incubated onto the compound scaffold at 3 passages. The adhesion, growth and differentiation of bone marrow mesenchymal stem cells on the compound scaffold were observed by the scanning electron microscope. Neuron-specific enolase was detected by immunofluorescence.
RESULTS AND CONCLUSION:The porosity and thermal sensitivity of HACC/β-GP scaffold and slow-release effect of glial cellline-derived neurotrophic factor were apparent. The results of MTT showed that the compound scaffold cannot take apparent negative effects to the proliferation of bone marrow mesenchymal stem cells. After inoculation, bone marrow mesenchymal stem cells permeated the porous structure of the scaffold and adhered to the scaffold. Under the role of glial cellline-derived neurotrophic factor, bone marrow mesenchymal stem cells showed neuron-like cellmorphology and cells co-cultured with the compound scaffold expressed the marker of neurons, neuron-specific enolase. Under the role of slow-release glial cellline-derived neurotrophic factor, bone marrow mesenchymal stem cells can grow wel in vitro and differentiate into neuron-like cells on the HACC/β-GP scaffold.

BACKGROUND:Exogenous neurotrophic factors or chemical induction can induce rat bone marrow mesenchymal stem cells to differentiate into neuron-like cells. However, exogenous inductors exert a short inducible action, and their chemical substances inevitably have a negative impact on cellviability to limit the application prospects of bone marrow mesenchymal stem cells to a certain extent.
OBJECTIVE:To investigate the effect of glial cellline-derived neurotrophic factor, green fluorescent protein gene transfection by adenovirus vector on biological characteristics of rat bone marrow mesenchymal stem cells, to observe the expression of glial cellline-derived neurotrophic factor and green fluorescent protein and the role of nutrition on bone marrow mesenchymal stem cells, and to explore the ability to differentiate into neuron-like cells induced by glial cellline-derived neurotrophic factor.
METHODS:The bone marrow mesenchymal stem cells at passage 3 were transfected by recombinant adenovirus (Multiplicity of infection=10, 50, 80, 100, 150, 200). The experiment had two groups according to target genes:bone marrow mesenchymal stem cells were transfected by Ad-GDNF-GFP in transfection group, and bone marrow mesenchymal stem cells were not transfected in control group. The expression of green fluorescent protein was detected by inverted fluorescence microscope. Transfection efficiency was calculated by flow cytometry. cells viability and the morphological changes of cells were compared respectively by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and inverted fluorescence microscope between the two groups. On days 5 and 10 after transfection, the expression of glial cel-derived neurotrophic factor mRNA was detected by PCR. On day 5, the expression of neuron-specific enolase was determined by immunofluorescence examination. On day 10, the expression of microtubule-associated protein 2 was identified.
RESULTS AND CONCLUSION:By the end of 12 hours after transfection, the green fluorescent protein expressed in cells, and the fluorescence intensity gradual y increased with time. When the multiplicity of infection was 100, the fluorescence intensity was strong and stable, and the transfection rate was nealy 90%on day 3 after transfection. cellviability in the transfection group was strengthened after transfection. On day 5 after transfection, bone marrow mesenchymal stem cells expressed neuron-specific enolase, and neuron-like protrusions gradual y extended. On day 10 after transfection, bone marrow mesenchymal stem cells expressed microtubule-associated protein 2 and glial cellline-derived neurotrophic factor mRNA, and exhibited neuron-like morphology and interconnected synpases. The recombinant adenovirus, Ad-GDNF-GFP, can highly transfect bone marrow mesenchymal stem cells when the multiplicity of infection is 100, and glial cellline-derived neurotrophic factor can promote the proliferation of bone marrow mesenchymal stem cells and induce bone marrow mesenchymal stem cells to differentiate into neuron-like cells.

Objective To explore the effects of different extracts of modified Siwu Siteng Decoction (TFT-Ⅰ, TFT-Ⅱ) on rat models with hyperuricemia. Methods Rat model with hyperuricemia was induced by hypoxanthine and oxonic acid potassium salt. Seventy rats were randomly divided into 7 groups:normal group, model group, allopurinol group, TFT-Ⅰ high and low dose groups, TFT-Ⅱhigh and low dose groups. Each dose group was given lavage with related medicine, and blank group and model group were given lavage with distilled water for one week, respectively. Uric acid in serum (SUA) was measured through fully automatic biochemical analyzer, and xanthine oxidase (XOD) activity in serum and liver tissue was measured through colorimetric method. Results Compared with model group, TFT-Ⅰ and TFT-Ⅱ high dose groups significantly reduced the level of SUA, serum and liver XOD activity (P<0.05), with significant statistical difference between the two groups (P<0.05). TFT-Ⅰ and TFT-Ⅱ low dose groups had no significant effects on SUA, and XOD activity in serum and liver (P>0.05). Conclusion Alcohol soluble ingredients of astragalus membranaceus and Siwu decoction in modified Siwu Siteng Decoction can effectively reduce the level of blood uric acid.

Lung cancer is a malignant tumor with high incidence rate and mortality rate in China and even the whole world, of which non-small cell lung cancer accounts for about 80%. Anaplastic lymphoma kinase (ALK) gene mutation accounts for about 5%. Alectinib, ALK-tyrosine kinase inhibitor (ALK-TKI), has great performance in clinical. The early detection and treatment of adverse drug reactions can greatly improve clinical benefits. This paper reports a patient of ALK positive non-small cell lung cancer was admited to Baotou Central Hospital in April 2020. The diagnosis and treatment was retrospectively analyzed, and the literature was reviewed.
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Anaplastic Lymphoma Kinase/genetics* ; Antineoplastic Agents/therapeutic use* ; Carbazoles/therapeutic use* ; Carcinoma, Non-Small-Cell Lung/secondary* ; Humans ; Lung Neoplasms/pathology* ; Mutation ; Piperidines/therapeutic use* ; Pleural Neoplasms/secondary* ; Protein Kinase Inhibitors/therapeutic use* ; Retrospective Studies ; Tomography, X-Ray Computed

Objective To investigate the characteristics of ocular surface inflammation index after pterygium excision by using Oculus Keratograph(R) 5M and Visual analogue scale and evaluated the effectiveness of antiinflammatory treatment.Methods A prospective case control study was performed.Eighteen patients (6 males and 12 females) who suffered from primary pterygium were recruited in Zhongshan Ophthalmic Center from June to September 2016.All patients were treated with monocular pterygium excision combined with amniotic membrane transplantation.Anti-inflammatory treatment was given after surgery,and the ocular inflammation index was evaluated at preoperative and 1st,3rd,7th,10th,30th and 60th day postoperative.The temporal conjunctival hyperemia index (TCHI) was assessed by Oculus Keratograph(R) 5M with a red eye index analysis software.Ocular symptom scores (OSS) and visual analogue scale (VAS) were used to analyze the subjective symptoms of the patients.Fluorescein staining was used to detect the epithelization of corneal and scleral wound.The best corrected visual acuity (BCVA),intraocular pressure and complications were evaluated in this study.This study was approved by the Medical Ethics Committee of Zhongshan Ophthalmic Center of Sun Yat-sen University (2016KYPJ024).All patients signed informed consent for clinical research.Results No drug-related ocular and systemic adverse events were found during the follow-up.Corneal epithelial defect was recovered on 10th day,and conjunctival epithelization was observed in sclera exposed area on 30th day.The BCVA on the 60th day was 0.12±0.17,which was significantly lower than 0.34±0.36 preoperatively (t =3.401,P =0.003).Compared with those before surgery,OSS and VAS were significantly increased on 1 st day (OSS:Z =-4.255,P =0.000;VAS:Z =-5.256,P =0.000).The OSS on 7th day was not significantly different from that before surgery (Z=-0.958,P=0.372).VAS decreased to baseline on 30th day.The OSS on 60th day after surgery was significantly lower than that before surgery (Z =-2.397,P =0.037).TCHI was higher than 1.2 preoperatively,and increased to the highest on 1 st day after surgery,with significant difference between them (t=-6.620,P=0.000).The TCHI decreased to baseline on 7th day,no significant difference were obtained when compared with preoperative TCHI (t =-1.050,P =0.310),and TCHI on 60th day after surgery was lower than that before surgery,with significant difference between them (t =2.758,P =0.020).Conclusions The subjective symptoms combined with conjunctival hyperemia can be more accurate assessment of ocular surface inflammation in the perioperative period of pterygium surgery,which can be used as an evaluation index to assess the effectiveness of anti-inflammatory treatment.

Objective To study the clinical efficacy and safety of icotinib hydrochloride tablets combined with whole brain radiotherapy (WBRT) in non-small cell lung cancer (NSCLC) brain metastasis.Methods Fifty-three patients with NSCLC brain metastasis who were unable to tolerate chemotherapy or chemotherapy failed were selected in Baotou Central Hospital from October 2010 to April 2015.The patients were divided into the observation group (n =27) and the control group (n =26) using random number table method.Two patients were dropped out,one in the control group and one in the observation group.The patients in the control group were given WBRT (30 Gy/15 Fx).On this basis,the patients in the observation group were given icotinib hydrochloride tablets 125 mg,3 times a day.The patients were followed up for 18 months after the end of WBRT.The adverse reactions,clinical efficacy and the median survival times of the two groups were compared.Results The objective response rate of the observation group was 88.5％,which was higher than that of the control group (64.0％),and the difference was statistically significant (x2 =4.238,P =0.040).The incidence rates of skin rash and liver function damage in the observation group were 76.9％ and 15.4％,which were higher than those in the control group (0,x2=31.638,P ＜ 0.001;x2 =4.173,P =0.041).However,most of them were degree Ⅰ-Ⅱ,and they were tolerable.There were no significant differences in the incidence of myelosuppression and gastrointestinal reactions between the two groups (26.9％ vs.20.0％,x2 =0.339,P=0.560;34.6％ vs.28.0％,x2 =0.259,P=0.611).The median survival time in the observation group (9.0 months) was longer than that in the control group (7.0 months).The one year survival rate of the observation group was 33.3％,which was higher than that of the control group (23.1％),but the difference was not significant (x2 =0.676,P =0.411).Conclusion Icotinib hydrochloride tablets combined with WBRT in the treatment of brain metastasis with NSCLC can significantly improve the curative effects of brain metastasis,and it has a survival advantage,with tolerable adverse reactions.

Objective:To compare the clinical value of stent assisted intestinal bypass and temporary loop ileostomy in laparoscopic low anterior resection of rectal cancer.Method:In this retrospective analysis, 57 patients undergoing laparoscopic low anterior resection for rectal cancer in the First Affiliated Hospital of Ningbo University from Jan 2020 to Jan 2022 were divided into intestinal bypass group (36 cases) and loop ileostomy group (21 cases).Result:There were no significant differences in postoperative GI function recovery and postoperative complication rate between the two groups (all P>0.05). The levels of albumin, prealbumin and hemoglobin in the intestinal bypass group were better than those in the ileostomy group when evaluated on 3rd months after operation [(40.5±2.3) g/L vs. (38.1±2.6)g/L、(26.4±2.7)mg/dl vs. (24.5±2.0)mg/dl、(137.6±5.9) g/L vs. (134.0±7.0) g/L, t=3.605、2.743、2.085, all P<0.05]. Hospital expenses of the intestinal bypass group was lower [(571 000±7 500) yuan vs. (69 300±9 100) yuan, t=-5.477, P<0.05]. Conclusion:Compared with traditional ileostomy, the stent assisted intestinal bypass reduces trauma with lower expenses and improves patients' status after laparoscopic low anterior resection for rectal cancer.