Genetically modified pigs with CD163 point mutation are resistant to HP-PRRSV infection.

Porcine reproductive and respiratory syndrome (PRRS) is a globally prevalent contagious disease caused by the positive-strand RNA PRRS virus (PRRSV), resulting in substantial economic losses in the swine industry. Modifying the CD163 SRCR5 domain, either through deletion or substitution, can eff1ectively confer resistance to PRRSV infection in pigs. However, large fragment modifications in pigs inevitably raise concerns about potential adverse effects on growth performance. Reducing the impact of genetic modifications on normal physiological functions is a promising direction for developing PRRSV-resistant pigs. In the current study, we identified a specific functional amino acid in CD163 that influences PRRSV proliferation. Viral infection experiments conducted on Marc145 and PK-15 CD163 cells illustrated that the mE535G or corresponding pE529G mutations markedly inhibited highly pathogenic PRRSV (HP-PRRSV) proliferation by preventing viral binding and entry. Furthermore, individual viral challenge tests revealed that pigs with the E529G mutation had viral loads two orders of magnitude lower than wild-type (WT) pigs, confirming effective resistance to HP-PRRSV. Examination of the physiological indicators and scavenger function of CD163 verified no significant differences between the WT and E529G pigs. These findings suggest that E529G pigs can be used for breeding PRRSV-resistant pigs, providing novel insights into controlling future PRRSV outbreaks.

Effect and Safety of Tongyan Spray () on Hyoid Motion in Patients with Dysphagia after Ischemic Stroke.

OBJECTIVE: To observe the effects and safety of Tongyan Spray () on the range and time of hyoid motion in patients with ischemic post-stroke dysphagia. METHODS: Seventy-two patients with ischemic post-stroke dysphagia were selected and randomly assigned to a treatment group (36 cases) and a control group (36 cases) by a random number table from January 2013 to October 2014. All patients swallowed 4 kinds of barium meals with different traits respectively, and each patient underwent video fluoroscopy (VF) examination twice. In the treatment group, Tongyan Spray was sprayed to the pharynx on both sides and the middle part once respectively. The spray was applied 30 min before the second examination. Purified water at room temperature was used as placebo in the control group. The changes in the range and time of hyoid motion in both groups were observed before and after treatment. RESULTS: Six patients dropped out in each group, and 60 patients completed the study and were included in the final analysis. Significant improvement was observed in the range of superior hyoid excursion distance and the time of hyoid motion in the treatment group compared with the control group (P<0.05). There were no obvious adverse reactions observed in oral mucosa in both groups during the whole study. CONCLUSION: Tongyan Spray was an effective and safe medicine for improving swallowing function in patients with ischemic post-stroke dysphagia.

Clinical study on tongyan spray for post-stroke dysphagia patients: a randomized controlled trial.

OBJECTIVE: To observe the effectiveness and safety of Tongyan spray composed of Chinese medicine for post-stroke dysphagia patients. METHOD: One hundred and twenty-two post-stroke dysphagia patients were randomly assigned to the treatment group (61 cases) and the control group (61 cases). Basic treatment was given to both groups, with Tongyan spray additionally used in oropharynx for the treatment group, and the placebo used for the control group. After 28-day treatment, the clinical effect and safety were evaluated according to the standard swallowing assessment (SSA) scale. RESULTS: One patient dropped out in each group, and 120 patients reached the final analysis of the study. The total effective rate for the treatment group was 71.7% (43/60), higher than 46.7% (28/60) in the control group (P<0.05), and the improvement on SSA scores of the two groups were significantly different after treatment (P<0.05). For grade 1 dysphagia patients (completely depending on nasogastric tube), the effective rate of the treatment group was 40.9% (9/22), and 12.5% (2/16) of the control group, without significant difference (P>0.05), while the improvement of SSA score was significantly different between the two groups after treatment (P<0.05). For grade 2-3 dysphagia patients (oral and nasogastric tube feeding), the total effective rate of the treatment group was 89.5% (34/38), higher than 59.1% (26/44) in the control group (P<0.05), and also the improvement on SSA scores was significantly different between the two groups after treatment (P<0.05). CONCLUSION: Tongyan spray was an effective and safe method for post-stroke dysphagia patients.

[Mode of debridement, negative-pressure therapy combined with tissue transplantation for treatment of complicated and refractory wounds].

OBJECTIVE: To observe therapeutic effect of negative-pressure treatment combined with tissue transplantation on complicated and refractory wounds after debridement. METHODS: After debridement, 20 patients with 20 complicated and refractory wounds hospitalized in our burn wards from May 2008 to June 2010 were randomly divided into treatment group (T, treated with negative-pressure from -19 kPa to -8 kPa, n = 10) and control group (C, covered with petrolatum gauze overlaid with saline gauze and dry gauze, n = 10) according to alternating method. On post treatment day (PTD) 4, 7, and 14, granulation tissues of wound surface in size of 4 mm × 3 mm × 2 mm were harvested for histopathological observation (including capillary growth, inflammatory cells, and collagen arrangement) with HE staining, and the numbers of vascular endothelial cells (VEC, with addition of rabbit anti-human coagulation factor VIII related antigen polyclonal antibody) and proliferation period cells (with addition of mouse anti-human Ki-67 monoclonal antibody) were counted by immunohistochemical staining. Data were processed with t test. Another 59 patients harboring 62 complicated and refractory wounds admitted to our burn ward at the same period were treated with the same mode of debridement, negative-pressure therapy, followed by timely skin or skin flap grafting. RESULTS: (1) Granulation tissue in T group grew more rapidly than that in C group. More capillaries and less inflammatory cells were observed in T group on PTD 7 as compared with those in C group. Collagen in T group on PTD 14 was more regular in arrangement than that in C group. The number of VEC per 400 times visual field in T group on PTD 4, 7, and 14 was respectively higher than that in C group (108.7 ± 11.2 vs. 31.0 ± 3.6, 138.0 ± 14.7 vs. 34.6 ± 4.5, 68.7 ± 6.9 vs. 55.1 ± 6.5, with t values from 4.62 to 30.28, P values all equal to 0.01). The number of proliferation period cell per 400 times visual field in T group on PTD 4 and 7 was respectively higher than that in C group (88.9 ± 5.9 vs. 16.6 ± 3.3, 128.1 ± 13.0 vs. 110.1 ± 8.9, with t value respectively 19.89, 3.33, P values all below 0.05). The number of proliferation period cell per 400 times visual field in T group on PTD 14 was obviously lower than that in C group (26.7 ± 5.1 vs. 59.7 ± 4.5, t = -12.43, P = 0.01). (2) After being treated with above therapeutic mode, necrotic tissues were removed completely and granulation tissue grew rapidly in 62 complicated and refractory wounds with high survival rate of skin grafts or skin flaps with good repair effect. CONCLUSIONS: Negative-pressure therapy can accelerate VEC formation and stimulate cell proliferation after debridement. Debridement, negative-pressure therapy, and timely skins/skin flaps grafting can effectively increase healing rate of complicated and refractory wounds.

[Study on differentiation of human umbilical cord-derived mesenchymal stem cells into human sweat gland cells in vitro and the relative signal pathway].

OBJECTIVE: To study the differentiation potential of human umbilical cord-derived mesenchymal stem cells (UCMSC) into human sweat gland cells (hSGC) and the role of extracellular signal-regulated kinase (ERK) pathway. METHODS: UCMSC and hSGC were isolated and cultured in vitro. The former was identified with expression of CD14, CD29, CD34, CD44, CD45, CD105, cytokeratin 7 (CK7), CK19, and carcinoembryonic antigen (CEA), while the latter was identified with expression of CK19 and CEA. UCMSC with density of 5 x 10(4) cells per well placed in lower compartment of Transwell chamber were divided into control group (C, cultured with nutrient solution without any stimulation), thermal injury group (TI, treated with heat-shocked hSGC with density of 1 x 10(4) cells per well inoculated into the upper compartment of Transwell chamber for indirect co-culture), thermal injury + EGF group (TIE, treated with indirect co-culture as used in TI group, with addition of 50 ng/mL EGF), thermal injury + PD98059 group (TIP, treated with indirect co-culture as used in TI group, with addition of 10 nmol/mL ERK specific inhibitor PD98059) according to the random number table. One week after culture, the positive expression rates of CK7 and CK19 in UCMSC were detected by flow cytometry, the expression of CK19 and CEA in UCMSC were examined with immunohistochemical staining and the positive expression rate of CEA was calculated, and the expression level of phosphorylated ERK (pERK) was determined by Western blotting. Data were processed with one-way analysis of variance. RESULTS: (1) CD29, CD44, and CD105 were highly expressed in UCMSC, accompanied by low or negative expression of CD14, CD34, CD45, CK7, CK19, and CEA. The expression of CK19 and CEA were positive in hSGC. The two results showed that UCMSC and hSGC were pure. (2) Compared with those of C group [(2.2 +/- 1.5)%, (2.2 +/- 0.7)%, (3.3 +/- 0.7)%, 0.640 +/- 0.026], the expression levels of CK7, CK19, CEA, and pERK in UCSMC of TI group [(6.4 +/- 0.7)%, (5.7 +/- 0.3)%, (7.4 +/- 1.0)%, 0.790 +/- 0.049] and TIE group [(14.3 +/- 1.0)%, (12.6 +/- 1.1)%, (17.6 +/- 2.3)%, 1.200 +/- 0.032] were significantly increased (with F value respectively 78.49, 139.36, 87.13, and 191.74, P values all below 0.01), and those of TIE group were higher than those of TI group (with F value from 50.14 to 145.47, P values all below 0.01). There were no obvious difference in the 4 indexes between TIP group and C group (with F value from 0.00 to 0.13, P values all above 0.05). CONCLUSIONS: UCMSC co-cultured with heat-shocked hSGC can differentiate into hSGC, and ERK signal pathway participates in the process of differentiation of UCMSC into hSGC.