Infectivity and antigenicity of pseudoviruses with high-frequency mutations of SARS-CoV-2 identified in Portugal.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has had a major impact on global human health. During the spread of SARS-CoV-2, weakened host immunity and the use of vaccines with low efficacy may result in the development of more-virulent strains or strains with resistance to existing vaccines and antibodies. The prevalence of SARS-CoV-2 mutant strains differs between regions, and this variation may have an impact on the effectiveness of vaccines. In this study, an epidemiological investigation of SARS-CoV-2 in Portugal was performed, and the VSV-ΔG-G* pseudovirus system was used to construct 12 spike protein epidemic mutants, D614G, A222V+D614G, B.1.1.7, S477N+D614G, P1162R+D614G+A222V, D839Y+D614G, L176F+D614G, B.1.1.7+L216F, B.1.1.7+M740V, B.1.258, B.1.258+L1063F, and B.1.258+N751Y. The mutant pseudoviruses were used to infect four susceptible cell lines (Huh7, hACE2-293T-293T, Vero, and LLC-MK2) and 14 cell lines overexpressing ACE2 from different species. Mutant strains did not show increased infectivity or cross-species transmission. Neutralization activity against these pseudoviruses was evaluated using mouse serum and 11 monoclonal antibodies. The neutralizing activity of immunized mouse serum was not significantly reduced with the mutant strains, but the mutant strains from Portugal could evade nine of the 11 monoclonal antibodies tested. Neutralization resistance was mainly caused by the mutations S477N, N439K, and N501Y in the spike-receptor binding domain. These findings emphasize the importance of SARS-CoV-2 mutation tracking in different regions for epidemic prevention and control.

Polyketone metabolites isolated from Rhodiola tibetica endohytic fungus Alternaria sp. HJT-Y7 and their SARS-CoV-2 virus inhibitory activitives.

Six new polyketone metabolites, compounds (1-6) and seven known polyketone compounds (7-13) were isolated from Rhodiola tibetica endophytic fungus Alternaria sp. The structural elucidation of five new polyketone metabolites were elucidated on the basis of spectroscopic including 2D NMR and HRMS and spectrometric analysis. Inhibition rate evaluation revealed that compounds 1(EC50 = 0.02 mM), 3(EC50 = 0.3 mM), 6(EC50 = 0.07 mM), 8(EC50 = 0.1 mM) and 9(EC50 = 0.04 mM) had inhibitory effect on the SARS-CoV-2 virus.

Design, Synthesis and Antiproliferative Evaluation of Novel Disulfides Containing 1,3,4-Thiadiazole Moiety.

A series of novel disulfides containing 1,3,4-thiadiazole moiety were designed, synthesized, and the structures of all products were identified by spectral data (IR, NMR, and high resolution (HR)-MS). Their in vitro antiproliferative activities were evaluated using 2-(2-methoxy-4-nitro-phenyl)-3-(4-nitro-phenyl)-5-(2,4-disulfopheyl)-2H-tetrazolium monosodium salt (CCK-8) assay against human cancer cell lines, A549 (human lung cancer cell), HeLa (human cervical cancer cell), SMMC-7721 (human liver cancer cell) and normal cell lines L929. The bioassay results indicated that most of the tested compounds 6a-k, 7a-k and 8a-k exhibited antiproliferation with different degrees, and some compounds showed better effects than positive control 5-fluorouracil (5-FU) against various cancer cell lines. Among these compounds, compound 6e exhibited the most potent inhibitory activity against A549 cells with IC50 value of 3.62 µM. Compounds 6i, 7a, 7g, 8a and 8b showed significantly antiproliferative activities against HeLa cells with IC50 values of 3.88, 3.76, 3.59, 3.38 and 3.12 µM, respectively. Compounds 6a, 7a and 8a owned high antiproliferative activities against SMMC-7721 cells with IC50 values of 2.54, 2.69 and 2.31 µM, respectively. Furthermore, all of the tested compounds showed weak cytotoxic effect against the normal cell lines L929. Based on the preliminary results, the substituent groups are vital for improving the potency and selectivity of this class of compounds.

[Benefit of post-circumcision application of skin wound induction gel in the treatment of phimosis in children].

OBJECTIVE: To study the effects of skin wound induction gel on the glans scabbing rate, class-A wound healing rate, and wound healing time of circumcision for phimosis in pediatric patients. METHODS: We randomly assigned 48 six to thirteen years old children with phimosis to an experimental group (n = 25) and a control group (n = 23) to be treated by circumcision. After surgery, the patients in the experimental group received application of skin wound induction gel while those in the control group received that of povidone iodine only to the glans and incision. We recorded and compared the glans scabbing rate, class-A wound healing rate, and wound healing time between the two groups of patients. RESULTS: Glans scabbing was observed in 3 cases in the experimental group and 17 cases in the control group (12.0% vs 73.9%, P < 0.01). No statistically significant differences were found in the rate of class-A wound healing between the two groups (100% vs 91.3%, P > 0.05). The wound healing time was significantly shorter in the experimental than in the control group ([10.7 ± 1.7] d vs [11.9 ± 2.1] d, P < 0.05). CONCLUSION: Post-circumcision application of skin wound induction gel to the glans and incision can effectively reduce glans secreta, alleviate inflammatory reaction, and shorten the healing time in the treatment of phimosis in children.

[Expression of hypoxia induced factor-1alpha and function of epididymis in varicocele: experiment with rats].

OBJECTIVE: To investigate the expression of hypoxia induced factor-la (HIF-1alpha) and the content of sialic acid and carnitine in the epididymis in varicocele (VC) and to investigate the mechanism of induction of epididymis dysfunction by VC. METHODS: Forty-five Wistar rats were randomly divided into 3 equal groups: experimental group undergoing decreasing of the diameter of the left renal vein so as to cause VC, control group, and sham operation group. 49 days after the operation the left epididymis samples were collected. Western blotting and immunohistochemistry were used to detect the HIF-1alpha expression. The contents of sialic acid and carnitine were detected. RESULTS: The HIF-1alpha expression level detected by Western blotting of the experimental group was 0.96 +/- 0.65, significantly higher than those of the control group (0.11 +/- 0.08) and sham operation group (0.07 +/- 0.05) (both P < 0.05); and the positive rate of HIF-1alpha detected with immunohistochemistry of the experimental group was 91.67%, significantly higher than those of the control group (16.67%) and sham operation group (21.43%) (both P < 0.01). The contents of sialic acid and carnitine in the epididymis of the experimental group were 6.3 +/- 2.3 and 1.1 +/- 0.3 respectively, both significantly lower than those of the control group (24.7 +/- 4.4 and 2.8 +/- 0.6) and sham operation group (26.1 +/- 4.4 and 3.2 +/- 0.6) (all P < 0.05). The contents of sialic acid and carnitine in the epididymis were significantly negatively correlated with the HIF-1alpha expression (r = -0.649, P = 0.017; r = -0.666, P = 0.013). CONCLUSION: VC causes epididymal hypoxia and epididymal dysfunction, and hypoxia plays an important role in VC induced epididymal dysfunction.

[Study on effect of sho-saiko-to compound on growth of nasopharyngeal carcinoma cells in CNE-bearing nude mice].

OBJECTIVE: To investigate the role of sho-saiko-to compound (SSTC) on the growth of the well-differentiated squamous cell line 1 of nasopharyngeal carcinoma (CNE-1) and well-differentiated CNE-2 in tumor-bearing nude mouse, and try to supply scientific data for its clinical development. METHOD: SSTC were prepared by concentration gradients, and the effect of SSTC on the growth and proliferation of the CNE-1 and CNE-2 were investigated by MT assay and soft-agar colony formation test. After setting up the subcutaneous tumor-bearing nude mouse model at the right lower back (0.2 mL CNE-2 cell suspension, 5 x 10(5)/mL), we randomly divided forty mice into 5 groups and gave high, middle and low concentration groups of SSTC (0.5, 0.25, 0.125 g X mL(-1) by intragastric administration. Positive and negative groups were set up for comparison. After constant administration for 15 days, the volume and weight of the tumor were measured for inhibition rate, so as to investigate the role of SSTC on the CNE-2 bearing tumor. RESULT: In vitro, compared with negative control, SSTC at different gradient concentrations were cultured with the CNE-1 and CNE-2 for 24 h, 48 h and 72 h. It showed that the growth and proliferation of both cell lines were inhibited to some extent. The inhibition rate was increased as the concentration and culture time increasing. Both MTT assay and soft-agar colony formation test showed that the 50% inhibiting concentration (IC50) was about 2.5 g X L(-1). In vivo, compared with negative control, the SSTC could slow down the tumor growth in the SSTC treated groups. The tumor growth of the negative control group (0.76 +/- 0.28) g, (962.88 +/- 245.96) mm3 and the low concentration group of SSTC (0.88 +/- 0.40) g, (1239.66 +/- 421.93) mm3 were obviously faster than those of the high, middle concentration group of SSTC (0.22 +/- 0.14) g, (239.31 +/- 137.07) mm3; (0.20 +/- 0.16) g, (263.42 +/- 166.57) mm3 and CTX positive control group (0.20 +/- 0.10) g, (246.72 +/- 194.6) mm3 (P<0.05). CONCLUSION: SSTC could efficiently inhibit the growth and proliferation of CNE-1 and CNE-2 in vitro, and slow down the tumor growth of the CNE-2 bearing nude mice. It may be a new compound of Chinese medicine for nasopharyngeal carcinoma therapy.

[Establishment of a new model of neurogenic bladder dysfunction caused by lumbar intervertebral diskhernia and its urodynamic evaluation].

OBJECTIVE: To establish a new rat model of neurogenic bladder dysfunction caused by lumbar intervertebral disk hernia, and to confirm the model by urodynamic examination. METHODS: Twenty male Wistar rats were divided into two groups at random:experimental group (n = 15) and pseudo-operation group (n = 5). The rats underwent laparotomy to disclose the intervertebral disk of L(6)-S(1), and a 1.50 mm x 4.50 mm blunt screw with flat end was inserted into the intervertebral disk of L(6)-S(1) of the rats in the experimental group so as to establish the model of lumbar intervertebral disk hernia. Computed radiography (CR) was performed 3 days after the operation to conform the successful insertion of the screw. Combined behavioral score (CBS) was used 1 d, 3 d, 1 week, 2 weeks, and 4 weeks after the operation. Four weeks after the laparotomy a vesical fistula above the pubis was made in all of the rats, and then urodynamic examination was performed three days after this operation. RESULTS: CR after operation confirmed that the blunt screw had been inserted into the lumbar disk of L(6). The CBS scores of the 2 groups at different time points all decreased along with time, and basically remained unchanged 1 week after. The CBS scores of the experiment group were significantly higher than those of the pseudo-operation group (all P < 0.05). The spontaneous vesical contraction rate in the filling period of the experimental group was (4.37 +/- 2.13) times/min, significantly higher than that of the pseudo-operation group [(0.06 +/- 0.13) times/min, t = 4.425, P = 0.000], the maximum bladder capacity of the experimental group was (1.20 +/- 0.34) ml, significantly greater than that of the pseudo-operation group [(0.60 +/- 0.14) ml, t = 5.141, P = 0.002], and bladder compliance of the experimental group was (0.024 +/- 0.012) ml/cm H(2)O, significantly lower than that of the pseudo-operation group [(0.096 +/- 0.088) ml/cm H(2)O, t = 2.891, P = 0.011], and the leak point pressure of the experimental group was (75 +/- 27) cm H(2)O, not significantly different from that of the pseudo-operation group [(62 +/- 23) cm H(2)O]. The urodynamic examination on the conscious rats confirmed the successful establishment of the neurogenic bladder dysfunction caused by lumbar intervertebral disk hernia. CONCLUSION: A new model of neurogenic bladder dysfunction caused by lumbar intervertebral disk hernia has been established by insertion of a blunt screw into the lumbar intervertebral disk of L(6). The model is confirmed by urodynamic examination.