We studied the function of connexin 43 (Cx43) gene in Xiao Meishan swine bone marrow mesenchymal stem cells (BMSCs) resulting from overexpression. Cx43 eukaryotic expression vector (pEGFP-Cx43) was constructed and transfected into BMSCs by nucleofector, after detecting the transfection efficiency; the expression of Cx43 was verified by RT-PCR, immunofluorescence and western blotting. Furthermore, we detected its cell cycle and apoptosis through flow cytometry. Our results show that pEGFP-Cx43 plasmid was successfully constructed, and green fluorescence in pEGFP-Cx43 transfected BMSCs was highly expressed with 60% transfection efficiency. In transgenic Xiao Meishan swines BMSCs, the expression level of Cx43 mRNA and protein were up-regulated. Meanwhile, the ability of cell proliferation was significantly increased, and the apoptosis rate was significantly reduced. Taken together, Cx43 overexpression could promote the proliferation of Xiao Meishan swine's BMSCs and markedly reduce their apoptosis, which provides evidence for in vivo research.
Animals ; Animals, Genetically Modified ; Apoptosis ; Cell Proliferation ; Connexin 43 ; metabolism ; Flow Cytometry ; Genetic Vectors ; Mesenchymal Stromal Cells ; metabolism ; Plasmids ; Swine ; genetics ; Transfection

Objective To investigate the efficacy and safety of balloon-assisted catheter directed thrombolysis (CDT) for acute lower extremity deep vein thrombosis (DVT).Methods From September 2008 to February 2011,94 patients with acute lower extremity DVT were admitted.The cases in early stage were treated by CDT (Group A,n =50),and the cases in late stage were treated by balloon-assisted CDT ( Group B,n =44).The clinical data of these patients were retrospectively analyzed.The circumference difference between normal and affected limbs,scores of venous patency,and rates of venous patency were recorded for judging the efficacy.The total dose of urokinase and retention time of infusion catheter was compared between the two groups.The incidence of pulmonary embolism and bleeding were used to judge the safety of treatment.The venous patency was followed up by ultrasound or/and venography.Measurement data with normal distribution was described by mean + standard,and was analyzed using T test.Measurement data with non-normal distribution was described by M ( QL,QU ),QL =P25,QU =P75,and was analyzed using Wilcoxon' s test.Categorical variable data was analyzed using Chi-Square test Results The prior treatment circunfference difference between normal and affectéd limbs were (5.37 ±1.97) cm (thigh) & (4.14 ± 1.57) cm (calf) in Group A and (5.41±2.22) cm (thigh) & (4.05 ±1.61) cm (calf) in Group B ; and the difference between the groups was insignificant ( thigh:t =- 0.113,P=0.910; calf:t =0.288,P =0.774).The post treatment correspondences were:(2.96 ± 1.10) cm (thigh) & ( 1.93 ± 0.84 ) cm (calf) in Group A and ( 1.78 ± 1.40) cm ( thigh ) & ( 1.41± 1.17 ) cm (calf) in Group B; the difference between the groups was significant (thigh:t =4.66,P ＜0.0001; calf:t =2.548,P =0.012 ).The prior treatment score of venous patency was 9 (8,10) in Group A and 8.3(7,10) in Group B without significant difference (Z =- 1.5172,P =0.1292).The post treatment score of venous patency was 3.5 ( 2,5 ) in Group A and 0 ( 0,1) in Group B with significant difference ( Z =-5.7702,P ＜0.01).The rate of venous patency after the treatment was 55.0％ (42.3％,72.4％ ) in Group A and 100％ (88.5％,100％ ) in Group B,with significant difference ( Z =4.9148,P ＜ 0.01).The total dose of urokinase used in the treatment was 5.950 ( 5.525,7.225 ) × 106U in Group A and 4.100 (3.600,5.050) × 106U in Group B with significant difference (Z =-6.0133,P ＜ 0.01).The retention time of perfusion catheter was 10 (9,12) d in Group A and 6 (5,7) d in Group B with significant difference ( Z =- 8.0358,P ＜ 0.01).No symptomatic pulmonary embolism occurred in both groups during the treatment and follow-up period.The rate of bleeding complication was 38.0％ (19/50) in Group A and 22.3％ (10/44) in Group B,without significant difference (x2 =2.5590,P =0.1097 ).The removal rate of optional filter was 88.37％ (38/43) in Group A and 100％ (39/39) in Group B,with significant difference ( x2 =4.829,P =0.028 ).The rate of venous patency at the last follow-up point was 50.0％ (44.4％,59.2％ ) in Group A,and 95.4％ (83.6％,100％ ) in Group B,with significant difference (Z =- 3.2721,P =0.0011).Conclusions Balloon-assisted CDT was a promising treatment for acute lower-extremity DVT.It improved the effect of thrombolysis and reduced the dosage of urokinase,and did not increase the risk of pulmonary embolism.

OBJECTIVE:To evaluate therapeutic efficacy of lamivudine(LAM)combined with adefovir dipivoxil(ADV) in the treatment of LAM-resistant chronic hepatitis B patients,and to study the relationship of therapeutic efficacy with hepatitis B virus(HBV)genotype. METHODS:A total of 101 LAM-resistant chronic hepatitis B patients selected during Dec. 2012 to Dec. 2014 were given LAM+ADV for 24 months at least. Regular outpatient visits or telephone follow-up were also performed. Polymerase chain reaction-reverse dot blot method was used to determine the HBV genotype. Chi-square test,Kaplan-Meier meth-od and Log-rank test were used to compare the virological response(HBV-DNA clearance rate)and biological response(ALT normalization rate and HBeAg seroconversion rate)among different genotypes at the 6th,12th,18th and 24th month of fol-low-up. RESULTS:The follow-up rate was 100%,without missed follow-up. Two genotypes were detected,including 34 pa-tients(33.7%)with genotype B and 67 patients(66.3%)with genotype C.At each time point mentioned above,the HBV-DNA clearance rates of 101 patients were 34.7%,55.4%,79.2% and 93.1%.At 6th month,HBV-DNA clearance rate and accumula-tive HBV-DNA clearance rate of genotype B were significantly higher than genotype C,with statistical significance(P<0.05). There was no statistical significance in HBV-DNA clearance rates or accumulative HBV-DNA clearance rates between different genotypes at other time points(P>0.05). At each time point mentioned above,ALT normalization rates of 101 patients were 40.6%,69.3%,82.2%,84.2%;there was no statistical significance in ALT normalization rates or accumulative ALT normaliza-tion rates between different genotypes(P>0.05). At each time point mentioned above,the HBeAg seroconversion rates of 101 patients were 10.9%,19.8%,24.8%,29.7%;there was no statistical significance in the HBeAg seroconversion rates or accu-mulative HBeAg seroconversion rates between different genotypes(P>0.05). CONCLUSIONS:LAM combined with ADV is ef-fective for LAM-resistant chronic hepatitis B patients. Moreover,the combination therapy can achieve earlier viological response in patients with genotype B than those with genotype C.

Clinical data of a child with leukoencephalopathy with brainstem and spinal cord involvement and lactate elevation (LBSL) diagnosed in the First Affiliated Hospital of Zhengzhou University in June 2020 were retrospectively analyzed.A female patient with 1 year and 2 months old presented with 10 months of developmental delay and 1 month of recurrent seizures.Physical examinations showed grade Ⅲ muscle strength of limbs, slightly higher muscle tone, active reflex of bilateral knee tendons, normal reflex of bilateral achilles tendons, and positive Babinski sign.Brain magnetic resonance imaging (MRI) showed bilateral cerebral hemisphere atrophy and diffuse abnormal signals.The whole exome sequencing test showed two heterozygous mutations of the DARS2 gene in the present case.There are no reports of early-onset LBSL gene mutation in Chinese population.A total of 6 related foreign literatures have been reported.All affected cases present psychomotor development disorders and other encephalopathy manifestations.Brain MRI involvement and DARS2 gene mutations are found in previous reports.Therefore, for children with developmental retardation, epilepsy, and diffuse abnormal signals in both cerebral hemispheres examined by brain MRI, genetic testing is recommended to confirm the diagnosis, guide prenatal diagnosis and genetic counseling.

Objective To evaluate the feasibility,safety and effectiveness of integrated treatmentwith comprehensively interventional techniques for acute deep venous thrombosis(DVT)of lower extremity. Methods During the period from June 2016 to June 2017,a total of 23 patients with acute DVT ofunilateral lower extremity were admitted to authors' hospital to receive integrated treatment. AngioJetthrombectomy,balloon angioplasty and iliac vein stent implantation were performed in the same setting.Angiography was performed immediately after the procedure. If the thrombus clearance rate was grade Ⅲand there were no large free clots(>5mm)in the lower extremity vein and inferior vena cava,the inferiorvena cava filters would be retrieved through their sheath,otherwise,the inferior vena cava filters would bereleased. All patients received continuous transcatheter infusion of urokinase until the thrombosis wasconfirmed to be completely dissolved. The patients were followed up at outpatient clinic at 1,3,6 and 12months after the treatment,and reexamination of color ultrasound and/or lower limb venography was used toassess the blood flow in the deep veins and in the stents. Results Angiography postoperatively showedthrombus clearance rate of grade Ⅲ was obtained in 20 patients(87%)who were successfully accomplishedintegrated treatment. The inferior vena cava filters had been retrieved after thrombolysis in 3 patients(13%)with thrombus clearance rate of grade Ⅱ. Twenty-one stents were implanted directly after thrombectomy in21 patients with iliac vein stenosis. There were no serious complications such as pulmonary embolism,severe hemorrhage,etc. Venous patency was confirmed by color ultrasound or lower limb venography. Conclusion Integrated treatment with comprehensively interventional techniques is feasible,safe,andeffective for acute DVT of lower extremity

Objective:To observe the effect of Buzhongyiqi Decoction on the expression of transforming growth factor β-3(TGFβ-3)and microRNA-30d(miR-30d)in uterosacral ligament tissues in a rat model of pelvic organ prolapse.Methods:According to the number lottery, 16 rats in the blank control group were divided into 2 groups: the A1 group(feeding 4 weeks, n=8)and the A2 group(undergoing the ovariotomy with normal saline gavage for 8 weeks, n=8). The 64 rat models of pelvic organ prolapse were randomly divided into 8 groups(8 rats in each group): the B1 group(feeding 4 weeks), the B2 group(given normal saline via gavage for 8 weeks), the C1 and C2 groups(given Buzhongyiqi Decoction via gavage, 3.5 ml·kg -1·d -1, for 4 and 8 weeks, respectively), the D1 and D2 group(given Buzhongyiqi Decoction via gavage, 7.0 ml·kg -1·d -1, for 4 and 8 weeks, respectively), the E1 and E2 group(given Buzhongyiqi Decoction via gavage, 14.0 ml·kg -1·d -1, for 4 and 8 weeks, respectively). Reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the expression of collagen COL1A1, COL3A1, TGF β-3 and miR-30d in uterosacral ligament tissues. Results:Compared with A1 group, the B1 group showed that the expression of COL1A1 and COL3A1 was decreased and the expression of TGFβ-3 and miR-30d was increased( P<0.01). After 4 weeks treament of Buzhongyiqi Decoction, the expression of COL1A1 was higher in E1 group than in C1 group ( P<0.01), The expression of COL3A1 in group D1 was singificantly higher than that in group C1, The expression of COL3A1 in group E1 was also singificantly higher than that in group D1 .The expression of TGF β-3 was higher in the D1 and E1 groups than in the C1 group( P<0.01). The expression of miR-30d was lower in the D1 and E1 groups than in the C1 group( P<0.01). After 8 weeks of Buzhongyiqi Decoction treatment, the expression of COL1A1 was higher in the D2 group than in the C2 group( P<0.01), while was lower in the E2 group than in the D2 group( P<0.01). The expression of COL3A1 and TGFβ-3 was higher in the D2 group than in the C2 group( P<0.01)and the expression of TGFβ-3 was lower in the E2 group than in the D2 group( P<0.01). The expression of miR-30d was lower in the D2 group than in the C2 group( P<0.01)and was higher in the E2 group than in the D2 group( P<0.01). Compared with the B1 group, the expression of COL1A1, COL3A1 and TGF β-3 was increased in the D1 and E1 groups( P<0.01), the expression of miR-30d was decreased in the C1, D1 and E1 groups( P<0.01). Compared with the B2 group, the expression of COL1A1 and COL3A1 was increased in the D2 and E2 groups, the expression of TGF β-3 was increased and the expression of miR-30d was decreased in the C2, D2 and E2 groups( P<0.01). Conclusions:Buzhongyiqi Decoction can increase the synthesis of collagen COL1A1 and COL3A1 in uterosacral ligament tissues of rats with pelvic organ prolapse, which may be related to the up-regulation of TGF β-3 expression and the down-regulation of miR-30d expression in rat uterosacral ligament tissues.

Objective To analyze the short-term survival and prognostic quality of life of patients with liver cirrhosis complicated by bacterial infection. Methods This study collected and analyzed 300 patients with liver cirrhosis complicated with infection who were hospitalized in the First Affiliated Hospital of Hebei North University, and followed up to discuss their survival and quality of life. Results In this study, the top two causes of infection were spontaneous bacterial peritonitis (60.67% of patients) and pneumonia (50.67% of patients). The second causes were urinary tract infections (15.33%), gastrointestinal infections (12.33%), and other causes. There was no statistically significant difference between male and female patients (P>0.05). In addition, the proportion of hospital infections was 71.00%, and there was no statistically significant difference between male and female patients (P>0.05). A total of 353 strains of pathogenic bacteria were isolated in this study (73.37% of patients with hospital infections). The distribution analysis of pathogenic bacteria showed that the highest proportion of ECO was 35.98%, followed by Klebsiella pneumoniae (18.98%). The distribution trend of 259 strains of pathogenic bacteria among hospital patients was consistent with that of all strains, and the difference was not statistically significant (P>0.05). Gram negative bacteria accounted for 79.60% (281/353) of all detected strains, of which Escherichia coli was mostly detected in patients with spontaneous bacterial peritonitis, Klebsiella Pneumoniae (KPN) and Pseudomonas aeruginosa (PAE) were mostly detected in patients with pneumonia, and Enterococcus (ENF) was mostly detected in patients with urinary tract infection; Among gram-negative bacteria, Staphylococcus epidermidis (SEP) and Staphylococcus aureus (SAU) are mostly found in patients with other infectious causes (blood flow infection, etc.) , and Streptococcus (STR) accounts for a high proportion in patients with Spontaneous bacterial peritonitis. In this study, 9 cases of death prognosis were detected during follow-up, and there was no statistically significant difference in the detection of death prognosis between different bacterial strains in both genders, as well as the difference in detection of death prognosis between hospital infections and out of hospital infections in both genders (P>0.05). There was no statistically significant difference in the detection of death prognosis between males and females due to different causes of infection, P>0.05. The quality of life scores of 291 surviving patients were compared between baseline and follow-up, indicating an increase in follow-up scores, especially in the dimensions of physiological function and physical pain. There was no statistically significant difference between different bacterial strains, infection causes, and hospital/non hospital infections (P>0.05) . Conclusion Spontaneous bacterial peritonitis and pneumonia are the main causes of infection that deserve special attention, and the main pathogens of infection are Gram negative bacteria. Targeted treatment and rehabilitation should be provided for patients with liver cirrhosis complicated by infection. At the same time, the proportion of hospital infections is relatively high, and attention should be paid to, prevention and control measures should be implemented as well.