Objective:To investigate the efficacy of oseltamivir combined with LianhuaQingwen granule in the treatment of influenza. Methods:Ninety-two patients with influenza who received treatment in Wenzhou People's Hospital from October 2017 to October 2019 were included in this study. They were randomly assigned to receive treatment with Lianhua Qingwen granules (control group, n = 46) or Lianhua Qingwen granules combined with oseltamivir (study group, n = 46) for 5 d. After treatment, clinical efficacy and symptom disappearance time (fever disappearance time, cough disappearance time, sore throat disappearance time) were compared between the control and study groups. Changes in immune function [C-reactive protein, interleukin-6, T lymphocyte subsets (CD 4+/CD 8+)] after treatment relative to before treatment, and the incidence of adverse reactions were compared between the two groups. Results:After treatment, total effective rate in the study group was significantly higher than that in the control group [97.83% (45/46) vs. 71.74% (33/46), χ 2 = 12.132, P < 0.001]. Fever disappearance time, cough disappearance time, sore throat disappearance time in the study group were significantly shorter than those in the control group [(2.04 ± 0.51) d vs. (3.15 ± 1.22) d, (3.25 ± 1.10) d vs. (4.53 ± 1.36) d, (2.72 ± 0.84) d vs. (3.95 ± 1.02) d, t = 5.693, 4.963, 6.313, all P < 0.001]. After treatment, C-reactive protein and interleukin-6 levels in the study and control groups were significantly decreased compared with before treatment [(9.24 ± 2.51) mg/L vs. (3.32 ± 2.21) mg/L, (9.21 ± 2.46) mg/L vs. (5.52 ± 2.10) mg/L, (70.25 ± 25.10) mg/L vs. (47.57 ± 14.50) mg/L, (69.24 ± 25.43) mg/L vs. (55.23 ± 16.44) mg/L]. After treatment, C-reactive protein and interleukin-6 levels in the study group were significantly lower than those in the control group ( t = 5.307, 3.138, all P < 0.001). After treatment, the number of CD 4+/CD 8+ cells in the study and control groups was significantly increased compared with before treatment [(0.45 ± 0.21) vs. (1.77 ± 0.59), (0.46 ± 0.18) vs. (1.08 ± 0.42)]. After treatment, the number of CD 4+/CD 8+ cells in the study group was significantly higher than that in the control group ( t = 14.295, 9.202, both P < 0.001). There were no significant differences in the incidence of nausea, vomiting, rash and diarrhea between the two groups within 5 d of treatment (all P > 0.05). Conclusion:Oseltamivir combined with Lianhua Qingwen granules exhibits better therapeutic effect than Lianhua Qingwen granules alone. It can strengthen immune function and shorten the durations of fever, cough and sore throat in patients with influenza.

BACKGROUND:The establishment of a good blood supply is a key mechanism for successful implantation of engineered tissues. OBJECTIVE:To observe the effect of basic fibroblast growth factor on the migration of human adipose-derived stem cells via implanting the human adipose-derived stem cells and sodium hyaluronate composite graft at the subcutaneous site of BALB/C mice, in order to explore an optimal scheme for soft tissue reconstruction. METHODS:Human adipose-derived stem cells were isolated from the adipose tissue of healthy cosmetic patients which received liposuction, and the cells were subcultured. Then 5×109/L passage 3 cellsuspension labeled by cm-dil was prepared. The working solution containing 2 mg/L basic fibroblast growth factor was prepared. Composite tissue al-lografts which were the mixtures of 0.25 mL sodium hyaluronate, 0.2 mL cellsuspension and 0.05 mL working solution or DMEM were implanted into the subcutaneous site of both sides of the mouse back. Specimens were taken at 6 weeks after operation and were evaluated histological y after hematoxylin-eosin and vascular immunofluorescent staining. RESULTS AND CONCLUSION:No necrosis, liquefaction, nodular tissue or gel remained in operated position. The hematoxylin-eosin staining showed the main components of the specimens were the adipose tissue and the loose connective tissue. The immunofluorescence staining showed the overlaps between the cm-dil fluorescence from human adipose-derived stem cells and the FITC fluorescence from the vascular endothelium in the experimental group were more than those in the control group (P<0.05). Basic fibroblast growth factor promotes the migration and the differentiation of human adipose-derived stem cells in the sodium hyaluronate scaffold into vascular endothelium.

BACKGROUND:Expanded polytetrafluoroethylene is a kind of porous polymer materials which is commonly used as clinical implants, and it has good biocompatibility, and is not easy to deformation or metamorphism. There is no existence of inflammation absorption reaction, and it al ows the cel migration and tissue ingrowth.
OBJECTIVE:To study the biocompatibility of expanded polytetrafluoroethylene scaffold and human adipose-derived stem cel s.
METHODS:The passage 4 human adipose-derived stem cel s were co-cultured with expanded polytetrafluoroethylene scaffold in vitro. The morphology and function of cel s adhered to the scaffold were observed by inverted phase contrast microscope, and cel adhesive rates and proliferation rates were also calculated by MTT assay.
RESULTS AND CONCLUSION:The inoculated cel s were round and bright, distributed on the surface of scaffolds uniformly, with good cel viability. After 3 hours a large number of adherent cel s were observed from the micrograph;after 24 hours there were a smal amount of short-spindle adipose-derived stem cel s. After cultured for 3 days, the short fusiform or polygon cel s could be seen clearly. After cultured for 7 days, the number of cel s increased significantly, few cel s fel off from the scaffold, and cel adhesion rate was up to an average of 95.7%. Meanwhile, the cel s revealed normal splitting proliferation rate. These findings indicate that human adipose-derived stem cel s are able to attach, grow and proliferate wel on the scaffold. Expanded polytetrafluoroethylene reveals excel ent cel ular compatibility and can be used as a vehicle for adipose tissue engineering.

Objective To investigate the expression and purification I278T-mutant human cystathionineβsynthase(CBS) in E . coli .Methods Site-directed mutagenesis by overlap extension using the polymerase chain reaction (PCR) was employed to construct mutant plasmids pGEX4T-1-CBS(I278T) ,which was induced and expressed in a medium containing 3% ethanol ,purified by affinity chromatography to obtain mutated CBS (I278T) protein .The activity ,UV-visible absorption spectroscopy ,protein particle size and Zeta potential of the purified protein were measured .Results Plasmid pGEX4T-1-CBS(I278T) was successfully constructed .The yield ,the specific activity and activity recovery of purified mutant CBS (I278T ) protein were 2 .3 mg/L ,21 .4 U/mg and 22 .6% .S-adenosylmethionine(AdoMet) with final concentration of 1 mmol/L showed no activation toward mutant CBS (I278T) protein .Ac-cording to UV-visible absorption spectroscopy analysis ,purified mutant CBS(I278T) had characteristic absorption peaks at 429 nm and 550 nm for heme-binding proteins .Protein average particle size was 7 .5 -10 .1 nm ,mainly in the form of tetramers ,and Zeta potential was - 16 .3 mV .Conclusion The methods of expression ,purification and identification of I278T-mutant human cystathionineβsynthase in E .coli were successfully established .

OBJECTIVETo discuss the effect of continuous positive airway pressure (CPAP) treatment for functional cardiac arrhythmias combined with obstructive sleep apnea hypopnea syndrome.

METHODSFifty-six OSAHS patients combined functional cardiac arrhythmia were randomized divided into two groups. The 28 patients in the control group were treated with metoprolol according to cardiac guidelines, the other 28 cases in the experimental group were treated with CPAP therapy combined with metoprolol. AHI and the lowest oxygen saturation (LSaO2) were tested before and after treatment.

RESULTSThe efficiency rates were 57.1% and 17.9% in experimental and control group respectively, with statistical difference (χ2 = 7.62, P < 0.01). Total effective rates were 85.7% and 53.6% respectively, with statistical difference (χ2 = 5.41, P < 0.05). In the experimental group, there were three treatment subgroups. After treatment, AHI and the lowest oxygen saturation were significantly different (P < 0.05).

CONCLUSIONCPAP treatment can effectively eliminate respiratory disturbance index, improve the symptoms of hypooxygen at night, and effectively improve the therapeutic effect of functional cardiac arrhythmias in OSAHS patients combined with functional arrhythmia.

Arrhythmias, Cardiac ; therapy ; Brugada Syndrome ; Cardiac Conduction System Disease ; Continuous Positive Airway Pressure ; Heart Conduction System ; abnormalities ; Humans ; Sleep Apnea, Obstructive ; therapy

Objective To study the effect of the exogenous recombinant human FGF-basic with different concentrations upon inducing human adipose-derived stem cells (hASCs) to differentiate into adipose cells,and the optimum concentration of exogenous rh-bFGF by experimental research.Methods hASCs were isolated and extracted by enzymatic digestion from the liposuction aspirate.hASCs using adipogenic supplement were divided into experimental group and blank group:the experimental group of adipogenic supplement was divided into adding the exogenous rh-bFGF 10 ng/ml,20 ng/ml and 40 ng/ml,the blank group of adipogenic supplement was cultured without exogenous rh-bFGF.MTT method was used to detect the adipocytes proliferation.The oil red O staining was used in the qualitative analysis on the time of newly forming adipocyte cells.Western blot was used to detect the effects of rh-bFGF on the expression of lipid droplets surface protein CIDEC at different stages during the culture.Results The experimental group could obviously shorten the period of inducing hASCs to differentiate into adioicytes,and promote the proliferation of adipocytes.The formation rate and the proliferation of adipocytes in the group adding 40 ng/ml rh-bFGF were superior to those in the experimental group else and blank group.The average time of the newly formed lipid droplets by adding 40 ng/ml rh-bFGFwas (11.5±1.9)h.The average absorbance of cell proliferation by adding 40ml rh-bFGF was 0.52 ±0.10.The CIDEC expression quantity of adding 40 ng/ml rh-bFGF group was also superior to that in the experimental group and blank group.Conclusions rh-bFGF in hASCs adipogenic supplement could promote the proliferation of adipocytes and dramatically accelerates the program of hASCs differentiating to adipocytes,in which the optimum concentration of rh-bFGF is 40ng/ml.

Objective To examine the prediction value of diffusion-weighted magnetic resonance imaging (DWI) on radiotherapy response in esophageal cancer.Methods A total of 24 subcutaneous esophageal cancer xenograft models were randomly divided into experimental group (n =14,received a single dose of 15 Gy radiotherapy) and control group (n =10,without any treatment).MRI were required before and after radiotherapy at different check time points (1,6,13 days) of T1WI,T2WI,and DWI measurements.Apparent diffusion coefficient (ADCX) and volume (VX) of each xenograft were measured,and both △ADCX and △VX were calculated.Results The ADC values of both group were decreased at the first day,however,the decrease in experimental group were more obviously with an increase at 6 and 13 d gradually.However,the ADC values of the control group showed a persistent decline.There was no significant difference in the ADC values between the two different groups before radiotherapy (P ＞ 0.05),while significant difference was found in the ADC values (F =6.178,16.181,58.733,P ＜ 0.05) and △ADC after radiotherapy (F =9.038,12.360,35.140,P ＜ 0.05).The xenografts volume in the experimental group showed a significant growth delay.There was no significant difference in volume between the two groups (P ＞ 0.05) before radiotherapy.Significant difference in V between the two groups only began to exist at 5 d after radiotherapy (F =28.587,P ＜ 0.05).The ADC0,ADC1 of transplanted tumor in control group had linear correlation relationships with its volume of later period.After radiotherapy,the trend of r values gradually increased from-0.118 to 0.896.Conclusions ADC values may change significantly at the early stage after radiotherapy,and initial and early ADC value may have close relationship with xenograft volumes of later period,which indicates that DWI has huge potential in the prediction of radiotherapy response.

Objective:To explore the differences of fecal calprotectin (FC), C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) between colon and small intestinal Crohn′s disease, and their predictive values for disease activity and mucosal healing in patients with small intestinal Crohn′s disease.Methods:From January 2017 to January 2023, 64 patients with Crohn′s disease who underwent capsule endoscopy in the First Affiliated Hospital of Zhejiang Chinese Medical University were enrolled, among them 28 patients had only small intestinal lesions (small intestine group) and 36 patients had lesions involving both small intestine and colon or only colon involvement (ileocolon group). The FC, CRP, and ESR levels of the two groups were detected and compared 15 days before capsule endoscopy examination. Wilcoxon rank-sum test was used for statistical analysis. Receiver operating characteristic curve analysis was used to evaluate the predictive value of FC, CRP, and ESR for disease activity and mucosal healing in patients with small intestinal Crohn′s disease.Results:The FC, CRP, and ESR levels of the small intestine group during the active phase of the disease were 1 689.00 μg/g (727.75 μg/g, 1 800.00 μg/g), 5.67 mg/L (1.00 mg/L, 17.01 mg/L), and 4.50 mm/1 h (2.00 mm/1 h, 11.00 mm/1 h), respectively; while FC, CRP, and ESR levels during the mucosal healing phase were 112.00 μg/g (46.50 μg/g, 130.50 μg/g), 1.00 mg/L (1.00 mg/L, 1.62 mg/L), and 2.00 mm/1 h (2.00 mm/1 h, 5.50 mm/1 h), respectively. The FC, CRP, and ESR levels of the ileocolon group during the active phase of the disease were 1 800.00 μg/g (895.50 μg/g, 1 800.00 μg/g), 4.94 mg/L (3.10 mg/L, 14.80 mg/L), and 10.00 mm/1 h (2.00 mm/1 h, 27.75 mm/1 h), respectively, while FC, CRP, and ESR levels during the mucosal healing phase were 66.00 μg/g (32.50 μg/g, 97.50 μg/g), 1.00 mg/L (1.00 mg/L, 1.55 mg/L), and 2.00 mm/1 h (2.00 mm/1 h, 4.50 mm/1 h), respectively. There were no statistically significant differences in FC, CRP, and ESR between the small intestine group and the ileocolon group during the active phase of the disease and mucosal healing phase (all P> 0.05). In the small intestine group, the levels of FC and CRP of patients during the active phase of the disease were 1 173.00 μg/g (312.00 μg/g, 1 800.00 μg/g) and 2.10 mg/1 L (1.00 mg/L, 16.00 mg/L), which were both higher than those of patients during the mucosal healing phase (112.00 μg/g (46.50 μg/g, 130.50 μg/g) and 1.00 mg/L (1.00 mg/L, 1.62 mg/L)), and the differences were statistically significant ( Z=-4.35 and-2.67, P<0.001 and =0.008). In the small intestine group, the level of ESR of patients during the active phase of the disease was 4.00 mm/1 h (2.00 mm/1 h, 16.00 mm/1 h), and there was no significant difference compared with that of patients during the mucosal healing phase (2.00 mm/1 h (2.00 mm/1 h, 5.50 mm/1 h)) ( P>0.05). When the cut-off level of FC was 188.50 μg/g, the sensitivity, specificity, and area under the curve for predicting disease activity in patients with small intestinal Crohn′s disease was 93.3%, 100.0%, and 0.964, respectively. When the cut-off value of CRP was 3.12 mg/L, the sensitivity, specificity, and area under the curve for predicting disease activity in patients with small intestinal Crohn′s disease was 46.7%, 92.3%, and 0.744, respectively. When the cut-off level of ESR was 10.00 mm/1 h, the sensitivity, specificity, and area under the curve for predicting disease activity in patients with small intestinal Crohn′s disease was 33.3%, 100.0%, and 0.654, respectively. There were no statistically significant differences in the area under the curve between the combinations of FC and CRP, FC and ESR, FC, CRP and ESR, and FC alone for predicting disease activity in patients with small intestinal Crohn′s disease (0.964, 0.959, and 0.959 vs. 0.964, all P> 0.05). There was a statistically significant difference in the area under the curve between the combination of CRP and ESR and FC alone in predicting disease activity in patients with small intestinal Crohn′s disease (0.708 vs. 0.964, Z=-2.57, P=0.010). Conclusions:There are no statistically significant differences in FC, CRP, and ESR between colon and small intestinal Crohn′s disease. FC has a high predictive value for disease activity and mucosal healing in patients with small intestinal Crohn′s disease and has certain clinical application value.

Objective:To investigate the clinical phenotype and genetic characteristics of infantile epileptic spasm syndrome caused by BRWD3 gene mutation. Methods:Clinical data of a child with BRWD3 related infantile epileptic spasm syndrome who was admitted to Department of Pediatric Neurology of Linyi People′s Hospital on August 2, 2019 were collected and followed up, whole exome sequencing technology and Sanger sequencing were applied to verify the child and his parents, and the pathogenicity of mutation site was analyzed. The studies till June 2023 were searched with keywords of " BRWD3" in both English and Chinese databases of China National Knowledge Infrastructure, Wanfang, Online Mendelian Inheritance in Man, and PubMed. The clinical phenotype and genetic characteristics of patients with BRWD3 related epilepsy were summarized. Results:The patient was a 4 years and 4 months old boy, with a clinical phenotype including severe global development delay, focal seizures (the onset age was 4 months), epileptic spasm (the onset age was 6 months), autism, megacephaly, high forehead as well as hypsarrhythmia. The whole exome sequencing results showed a de novo and frameshift variation c.4318_4319del(p.Q1441Efs*20)(NM_153252) in the BRWD3 gene, and the variation was interpreted as pathogenic (PVS1+PS2+PM2) according to the American College of Medical Genetics and Genomics variant classification criteria and guidelines. A total of 7 English literature articles were retrieved reporting 16 cases of BRWD3 gene related epilepsy in children (including 1 case of infantile epileptic spasm syndrome), and there has been no report in China yet. Totally there were 17 cases of BRWD3 gene related epilepsy including this case. All the cases showed X chromosome dominant inheritance, of whom 15 cases showed minor variations, including 7 missense variations, 3 frameshift variations, 3 splicing variations, 2 nonsense variations, and the remaining 2 cases showed large segment deletions. A total of 15 different variants were found. The phenotypes of the 17 patients mainly included epileptic seizures (17/17), intellectual disability (10/17), motor development disorder (7/17), speech impairment (9/17), megacephaly (8/17), facial malformation (8/17), autism (4/17) and hypotonia (4/17). The common seizure types were found to be focal seizures, occasionally epileptic spasm seizures and tonic seizures. Conclusions:BRWD3 gene variation related epilepsy is an X chromosome dominant genetic disease with a wide clinical phenotype spectrum. BRWD3 gene mutation c.4318_4319del(p.Q1441Efs *20) could cause infantile epileptic spasm syndrome, manifested as severe global developmental delay, epileptic spasm, focal seizures, autism, craniofacial malformation and hypsarrhythmia. This research enriches BRWD3 gene mutation spectrum.

Objective:To evaluate the impact of the reform of the county medical community on the expenditure of medical insurance funds, and to provide references for maintaining the stability of the medical insurance fund and deepening the reform of the medical community.Methods:Medical insurance data of urban and rural residents in M County, Yunnan province from 2016 to 2019 were collected, and a discontinuous time series model was used to analyze the impact of county medical community reform on medical insurance fund expenditures.Results:Since the reform, the number of patients discharged from county-level hospitals has shown a downward trend, averaging a decrease of 25.996 people per month; yet increases have emerged with the average hospitalization expenditure, the average hospitalization overall fund expenditure, and the average outpatient overall fund expenditure, averaging a monthly increase of 90.931 yuan, 50.014 yuan and 1.528 yuan respectively. The average hospitalization expenditure, the average hospitalization overall fund expenditure, and the average outpatient overall fund expenditure of the township hospitals all showed an upward trend, averaging a monthly increase of 31.191 yuan, 38.678 yuan and 0.085 yuan respectively. The flow of external medical insurance funds of the medical community has shown a continuous upward trend, averaging a monthly increase of hospitalization fund expenditures of 33.005 yuan, and a monthly increase of outpatient overall fund expenditures of 4.896 yuan overall.Conclusions:The M County medical community should further strengthen the top-level design, explore the reform of medical insurance payment methods, improve the regional information platform, standardize the referral system, and strengthen supervision to deepen the construction of the medical community to ensure the sustainable operation of medical insurance funds.