[Identification and preservation of arm lymphatics in axillary lymph node dissection to prevent arm lymphedema: a single center randomized controlled trial].

Objective: To evaluate the feasibility of identification and preservation of arm lymphatics (DEPART) in axillary lymph node dissection (ALND) for breast cancer to prevent arm lymphedema. Methods: A randomized controlled study method was used. Two hundred and sixty-five patients who underwent breast cancer surgery at the Department of Thyroid and Breast Surgery, Zhongnan Hospital of Wuhan University from November 2017 to June 2018 were included, and the patients were randomly divided into ALND+ DEPART group (132 patients) and standard ALND group (133 patients) by random number table method. In the ALND+ DEPART group, indocyanine green and methylene blue were injected as tracers before surgery, and the arm sentinel nodes was visualized by staged tracing during intraoperative dissection of axillary lymph nodes. Partial frozen sections were made of arm lymph nodes >1 cm in length and hard and suspicious of metastasis, and arm lymph nodes and lymphatic vessels were selectively preserved. Patients in the standard ALND group underwent standard ALND. Objective and subjective indexes of arm lymphedema were evaluated by 5-point circumference measurement and Norman questionnaire. Results: Among 132 breast cancer patients in the ALND+ DEPART group, 121 (91.7%) completed DEPART. There were no statistically significant differences in age, body mass index, pathological type, dissection number of axillary lymph node, N stage, TNM stage, molecular typing, and regional radiotherapy between the ALND+ DEPART and standard ALND groups (P>0.05). At a median follow-up of 24 months, assessment by the 5-point circumference measurement showed that the incidence rates of lymphedema in the ALND+ DEPART and standard ALND groups were 5.0% (6/121) and 15.8% (21/133), respectively, with statistically significant differences (P=0.005). Assessment by the Norman questionnaire showed that the incidence rates of lymphedema in the ALND+ DEPART and standard ALND groups were 5.8% (7/121) and 21.8% (29/133), respectively, with a statistically significant difference (P<0.001). No local regional recurrence was observed in either group during the follow-up period. Conclusion: For breast cancer patients with positive axillary lymph nodes, the administration of DEPART during ALND can reduce or avoid the occurrence of arm lymphedema without compromising oncology safety.

[Lesion removal plus whole breast exploration and washing plus micro-plastic procedures in the treatment of granulomatous lobular mastitis: a randomized controlled study].

Objective: To examine the effect of"lesion removal plus whole breast exploration and washing plus micro-plastic surgery"in granulomatous lobular mastitis. Methods: A single-center prospective randomized controlled study method was used to enroll patients diagnosed with granulomatous lobular mastitis for whom surgical procedures were projected from March 2017 to September 2019 at Department of Thyroid and Breast Surgery, Zhongnan Hospital of Wuhan University. The sample size is determined by the superiority test. Based on the literatures and the previous work, the two groups require 97 cases. Fifty-two patients underwent"lesion removal+whole breast exploration and washing plus micro-plastic surgery"(observation group). Forty-five cases underwent"empirical breast lesion resection plus fascia tissue flap plasty plus nipple and areola correction"(control group). The primary study endpoint is the recurrence rate, and the secondary study endpoints include surgical complications, incision healing time, and postoperative patient satisfaction. Independent sample t test, Wilcoxon rank-sum test, χ² test and Fisher exact test were used for comparison between groups. Results: All procedures were completed successful, with no severe complications. All patients were followed up for (15.2±1.9) months (range: 12 to 24 months). There were no significant differences in incidence of postoperative complications (7.7% (4/52) vs. 6.7%(3/45), P=1), drainage time ((8.6±0.6) days vs. (8.4±0.8) days, t=1.921, P=0.053) and hospital stay ((7.7±0.6) days vs. (7.6±0.5) days, t=1.633, P=0.102) between the two groups. The recurrence rate of the observation group was lower significantly than that of the control group (3.8% (2/52) vs. 24.4%(11/45), χ²=8.819, P=0.003). The observation group had better cosmetic effects (Z=-2.657, P=0.008) and patient satisfaction than control group (Z=-5.730, P=0.000). Conclusion: The "lesion removal plus whole breast exploration and flushing plus micro-plastic surgery" has a good therapeutic effect and cosmetic value for patients with refractory granulomatous lobular mastitis.

FBXO17 promotes malignant progression of hepatocellular carcinoma by activating wnt/ß-catenin pathway.

OBJECTIVE: This study was to investigate the expression of FBXO17 in hepatocellular carcinoma (HCC) and its relationship with clinical HCC features and patient prognosis. PATIENTS AND METHODS: The expression of FBXO17 at mRNA level and protein level in tumor tissues and paracancerous tissues of 45 patients with HCC was respectively detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot. Besides, FBXO17 expression and its pathological characteristics of HCC, as well as the prognosis of patients, were also analyzed. Then, the expression level of FBXO17 in HCC cell lines was further verified using qRT-PCR assay. In addition, FBXO17 overexpression and knockdown models were constructed using lentivirus in HCC cell lines including Bel-7402 and HepG2. Cell counting kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EDU), cell clone experiment, flow cytometry, and transwell assay were used to explore the effect of FBXO17 on the biological function of HCC cells. Finally, whether FBXO17 could exert its biological characteristics through wnt/ß-catenin pathway was determined. RESULTS: Results showed that FBXO17 expression in tumor tissues of HCC patients was markedly higher than that in adjacent tissues. Meanwhile, compared with patients with low FBXO17 expression, the pathological grade was higher and the overall survival rate was lower in patients with high expressed FBXO17. In vitro experiments showed that the cell proliferation and metastasis ability in the Anti-FBXO17 group was markedly decreased, and the apoptosis was significantly enhanced compared with the NC group. In contrast, overexpression of FBXO17 markedly increased cell proliferation and metastasis ability while decreased cell apoptosis. Finally, Western blot results indicated that silencing FBXO17 might function through downregulating the expression of proteins in wnt/ß-catenin pathway such as c-Myc, MMP-9, and MMP-2 while upregulating GSK-3ß level, thereby promoting the malignant progression of HCC. CONCLUSIONS: FBXO17 was significantly increased in tumor tissues of HCC patients, which was significantly associated with pathological stage and poor prognosis of these patients. In addition, FBXO17 might promote the malignant progression of HCC by inhibiting wnt/ß-catenin pathway.

[Efficacy of Pipeline embolization device in the treatment of large and giant intracranial aneurysms].

Objective: To summarize the safety and efficacy of Pipeline embolization device in the treatment of large and giant intracranial aneurysms (maximum diameter ≥ 10 mm). Methods: The clinical and imaging data of 22 patients with intracranial aneurysms treated by Pipeline Embolization Device (PED) from February 2015 to July 2016 in the Zhengzhou University People's Hospital were retrospectively analyzed. The results about postoperative clinical and imaging follow-up and complications were evaluated. Results: The PED were successfully implanted in 22 patients with 22 intracranial aneurysms. Two aneurysms were treated with PED alone, and 20 aneurysms were treated with PED assisted coil embolization. Perioperative complications occurred in 3 patients (13.6%), and 1 patient died because aneurysm ruptured 3 days after operation. Nineteen patients (19 aneurysms) obtain imaging follow-up visit 3-17 months after operation, 13 of the 19 aneurysms (68.4%) received complete and subtotal occlusion, with no aneurysm recurrence. Eighteen patients obtain clinical follow-up visit 16-32 months after operation, with mRS (modified Rankin scale) score 1 in 5 patients, mRS score 0 in 13 patients, none developed new neurologic symptoms. Conclusions: PED has showed the high rate of obliteration for the treatment of large and giant intracranial aneurysms. But the complications needs to be further studied.

Epidemiological and morbidity assessment of Schistosoma japonicum infection in a migrant fisherman community, the Dongting Lake region, China.

We assessed the epidemiology and morbidity related to Schistosoma japonicum infection by both parasitological (Kato-Katz method and miracidium hatching test) and ultrasound examination in a migrant fisherman community (n = 106) from the Dongting Lake region in China in 2001. A prevalence of 69.8% and a mean infection intensity of 66.5 eggs per gram (epg) were recorded for this group. Males had a higher level of infection (77%) and intensity (92.2 epg) compared with females (58% and 28.6 epg, respectively). Subjects aged 11-20 years had the highest prevalence of infection (91%) and the heaviest infection intensity (85.3 epg) among all age groups. Twenty-six percent of the subjects had not been treated previously for schistosomiasis. Parenchymal fibrosis (> or = stage II) detected by ultrasound and spleen enlargement (or splenectomy) occurred in 37% and 9% of subjects, respectively. Portal vein dilation was detected in 18% subjects, and 58% (11/19) of those patients with portal vein enlargement were shown to have parenchymal fibrosis (> or = stage II). These results indicate that S. japonicum infection and related morbidity in mobile fisherman is still unacceptably high despite the fact that China has implemented a Schistosomiasis Control Project financed by a World Bank Loan for a period of 8 years since 1992. Clearly, new approaches for schistosomiasis control, especially in communities like these mobile fishermen, should be explored.

Field assessment of recombinant Schistosoma japonicum 26 kDa glutathione S-transferase in Chinese water buffaloes.

We have shown previously that anti-fecundity immunity can be induced experimentally against recombinant 26 kDa glutathione S-transferase (reSjc26GST) in Chinese water buffaloes (Bos buffelus), important reservoir hosts for Schistosoma japonicum in China. In the field study described here, we immunized buffaloes with reSjc26GST to induce protective immunity against S. japonicum and to evaluate its effectiveness in controlling schistosomiasis japonica. We selected two villages as test and control groups in inside-embankment areas endemic for schistosomiasis japonica. The buffaloes in the test village were vaccinated with reSjc26GST, whereas those in the control village were not. The indicators of the effect of the vaccine included the generation of specific IgG antibodies in the vaccinated buffaloes, changes in the prevalence and infection intensity in buffaloes and village children, changes in the density of infected snails, and changes in the infectivity of water bodies (assessed by sentinel mice) in transmission areas adjacent to both villages. Twenty months after vaccination, the infection rate of buffaloes in the test village was decreased by 60.4% (from an initial prevalence of 13.5% to 5.4%), and 67.9% when compared with that in the control village (initial prevalence of 16.7%). However, the infection rate in village children remained unchanged. The density of infected snails decreased by 71.4%, from 0.0049/0.11 m2 to 0.0014/0.11m2 in the high transmission area outside the embankment in the test village. There was no change in the infectivity of the water body transmission areas between the test and control villages. The levels of specific antibodies to reSjc26GST showed a continuous increase after vaccination. These results indicate that protective immunity was induced and maintained in buffaloes after vaccination with reSjc26GST. The vaccine could thus play a significant role in reducing S. japonicum transmission caused by water buffaloes in the Lake region of China.

A comparative study of third-order nonlinear optical properties of silver phenylacetylide and related compounds via ultrafast optical Kerr effect measurements.

A comparative study of the third-order nonlinear optical properties, via the newly developed heterodyned optical Kerr effect (OHD-OKE) measurements, of silver phenylacetylide and related compounds is reported. [AgC[triple bond]CC(6)H(5)](n) (1) was found to exhibit efficient third-order nonlinear optical susceptibility chi((3)) of 2.4 x 10(-14) esu, and second hyperpolarizability gamma of 9.07 x 10(-32) esu. These results are compared with those of two related silver phenylacetylide compounds, namely, a double salt, (silver phenylacetylide).(silver tert-butylthiolate) [AgC[triple bond]CC(6)H(5).AgS(t-C(4)H(9))](n) complex (2), and a cluster, triphenylphosphine silver phenylacetylide tetramer, [(C(6)H(5))(3)PAgC[triple bond]CC(6)H(5)](4) (3), as well as that of the related organic polymer polyphenylacetylene (4). These four compounds represent different types of phenylacetylide derivatives: 1 is an organometallic polymer, 2 a polymeric double salt, 3 a discrete metal cluster, and 4 an organic polymer. It was found that the third-order optical nonlinear response was enhanced by the incorporation of silver d electrons into the delocalized conjugated organic pi system, and its magnitude is highly dependent upon the extent of the pi delocalization. Specifically, the relative magnitudes of chi((3)) and gamma follow the order silver phenylacetylide polymer (1) > (silver phenylacetylide).(silver tert-butylthiolate) double salt (2) > polyphenylacetylene polymer (4) > tetrameric (triphenylphosphine silver phenylacetylide)(4) cluster (3). The observed trend may be attributed to the decreasing length of pi conjugation. It is interesting to note that the incorporation of Ag(I) into the polymeric framework of polyphenylacetylene enhances the chi((3)) by 25-fold for the same degree of polymerization (n = 7). The signs of chi((3)) and gamma, which are related to the response mechanisms, were found to be solvent dependent.

Differential mercury volatilization by tobacco organs expressing a modified bacterial merA gene.

Mercury pollution is a major environmental problem accompanying industrial activities. Most of the mercury released ends up and retained in the soil as complexes of the toxic ionic mercury (Hg2+), which then can be converted by microbes into the even more toxic methylmercury which tends to bioaccumulate. Mercury detoxification of the soil can also occur by microbes converting the ionic mercury into the least toxic metallic mercury (Hg0) form, which then evaporates. The remediation potential of transgenic plants carrying the MerA gene from E. coli encoding mercuric ion reductase could be evaluated. A modified version of the gene, optimized for plant codon preferences (merApe9, Rugh et al. 1996), was introduced into tobacco by Agrobacterium-mediated leaf disk transformation. Transgenic seeds were resistant to HgCl2 at 50 microM, and some of them (10-20% ) could germinate on media containing as much as 350 microM HgCl2, while the control plants were fully inhibited or died on 50 microM HgCl2. The rate of elemental mercury evolution from Hg2+ (added as HgCl2) was 5-8 times higher for transgenic plants than the control. Mercury volatilization by isolated organs standardized for fresh weight was higher (up to 5 times) in the roots than in shoots or the leaves. The data suggest that it is the root system of the transgenic plants that volatilizes most of the reduced mercury (Hg0). It also suggests that much of the mercury need not enter the vascular system to be transported to the leaves for volatilization. Transgenic plants with the merApe9 gene may be used to mercury detoxification for environmental improvement in mercury-contaminated regions more efficiently than it had been predicted based on data on volatilization of whole plants via the upper parts only (Rugh et al. 1996).

HAL1 mediate salt adaptation in Arabidopsis thaliana.

The yeast HAL1 gene was introduced into Arabidopsis thaliana by Agrobacterium tumefaciens-mediated transformation with vacuum infiltration under the control of CaMV 35S promoter. Thirty-three individual kanamycin resistant plants were obtained from 75,000 seeds. Southern blotting analysis indicated that HAL1 gene had been integrated into all of the transgenic plants' genomes. The copy number of HAL1 gene in transgenic plants was mostly 1 to 3 by Southern analysis. Phenotypes of transgenic plants have no differences with wild type plants. Several samples of transformants were self-pollinated, and progenies from transformed and non-transformed plants (controls) were evaluated for salt tolerance and gene expression. Measurement of concentrations of intracellular K+ and Na+ showed that transgenic lines were able to retain less Na+ than that of the control under salt stress. Results from different tests indicated the expression of HAL1 gene promotes a higher level of salt tolerance in vivo in the transgenic Arabidopsis plants.

[Screening and cloning of genes encoding Schistosoma japonicum antigens related to the serum antibodies in Mirotus fortis].

OBJECTIVE: To understand and identify the molecules related to the natural resistance to Schistosoma japonicum infection in Mirotus fortis. METHODS: Sera from Mirotus fortis without schistosome infection were collected. The S. japonicum adult worm cDNA library was immunologically screened with the sera. The positive recombinants were identified, cloned, sequenced and analysed with software and internet. RESULTS: Seven genes encoding antigens relevant to sera antibodies in Mirotus fortis were cloned and sequenced. These antigens included glyceraldehyde 3-phosphate dehydrogenase (GAPDH), serine protease inhibitors (SERPIN), 70 kDa heat shock protein (HSP70), 22.6 kDa membrane-associated antigen, paramyosin (Sj97), cytochrome C and cathepsin B. CONCLUSION: Many protein molecules might have been involved in natural resistance to S. japonicum infection in Mirotus fortis. The above 7 kinds of molecules may be identified as new candidates of vaccine against S. japonicum infection.

[A study on the recombinant 26 kDa glutathione-S-transferase as a vaccine candidate: dynamics of antibodies in immunized buffaloes and protection against Schistosoma japonicum infections].

OBJECTIVE: To observe the dynamics of antibodies and protection against Schistosoma japonicum infections in buffaloes after immunized with recombinant 26 kDa glutathione-S-transferase (reSjc26GST). METHODS: Buffaloes in 2 villages endemic for schistosomiasis japonica were selected as test and control groups, respectively. In test group initially 96 buffaloes were vaccinated with reSjc26GST, and 90 buffaloes in the control group did not experience vaccination. The indicators included levels of antibodies to reSjc26GST in buffaloes before and after infection with S. japonicum and changes in infection rate. RESULTS: Specific antibodies, which showed a trend of trapezoid increase, were induced in buffaloes after immunized with reSjc26GST. Twenty months after immunization, the infection rate of the test group was decreased by 62.2% when compared with that before vaccination, and by 67.7% when compared with that of the control in the corresponding period. CONCLUSION: Specific antibodies and a certain extent of protection were induced in buffaloes after immunized with reSjc26GST, which played an significant role in ameliorating morbidity.

Leafy head formation of the progenies of transgenic plants of Chinese cabbage with exogenous auxin genes.

The experiment was performed to evaluate the progenies of plant lines transgenic for auxin synthesis genes derived from Ri T-DNA. Four lines of the transgenic plants were self-crossed and the foreign auxin genes in plants of T5 generation were confirmed by Southern hybridization. Two lines, D1232 and D1653, showed earlier folding of expanding leaves than untransformed line and therefore had early initiation of leafy head. Leaf cuttings derived from plant of transgenic line D1653 produced more adventitious roots than the control whereas the cuttings from folding leaves had much more roots than rosette leaves at folding stage, and the cuttings from head leaves had more roots than rosette leaves at heading stage. It is demonstrated that early folding of transgenic leaf may be caused by the relatively higher concentration of auxin. These plant lines with auxin transgenes can be used for the study of hormonal regulation in differentiation and development of plant organs and for the breeding of new variety with rapid growth trait.

Molecular characters and morphological genetics of CAL gene in Chinese cabbage.

BcpCAL, the homologous gene of CAL, was isolated from Chinese cabbage. Unlike BobCAL of cauliflower, BcpCAL did not hold the terminating mutation in the fifth exon. After crosses of cauliflower with Chinese cabbage, the resultant hybrids failed to form curd, which implicates the genetic complement of BcpCAL to the mutated BobCAL in the function of curd formation. One of CAL gene isolated from the hybrid apparently comes from the female parent (Chinese cabbage) even though there are a few of the bases substituted and deleted. The result offers the molecular and genetic evidences for the study of biological function of CAL in morphological genetics of curd.

[Preliminary report on immunoscreening of Schistosoma japonicum adult worm cDNA library by sera from infected Microtus fortis].

Microtus fortis (Mf) is resistant to Schistosoma japonicum (Sj) infection. To explore if Mf responses to special Sj antigens after infection, Sj adult worm cDNA library was screened with sera from infected Mf. Twelve positive clones were obtained after rescreening. After automatically excised with the helper phage, positive clones were further identified by PCR. The size of Sj cDNA fragments in positive clones ranged from 300 bp to 1.8 kb. Six of these fragments were 300 bp, one was 1 kb, and five were 1.8 kb in size. The results indicate that the infected Mf serum could recognize Sj specific antigens. It is valuable to further study the immune protection of these cloned molecules.

[Effects of IAA and stimulated microgravity on formation of adventitious roots of Chinese cabbage].

Under the induction of indole-3-acetic acid (IAA), adventitious roots were differentiated on hypocotyl segments derived from seedlings of Chinese cabbage (Brassica campestris spp. pekinensis). IAA at concentration of 0.4-1.0 mg/L in solid MS medium incited many adventitious roots on hypocotyl segments. The earliest anatomic changes were observed on cut surface of hypocotyl segments under optical microscope 24 hours after IAA treatment: cytoplasmic and nuclear density became higher in a few of parenchytmatous cells adjacent to phloem in tissue of pericycle, followed by cell divisions. Lately, the dividing cells expanded and developed into root primordium from which root cap was differentiated. After five days, most roots protruded through hypocotyl cortex and appeared just below the cut surface. The rooting capacity of the segments derived from three regions of each hypocotyl was different. High level of IAA modified the polarity of root formation on segment inserted upside down and sucrose increased the function of IAA. Additionally, microgravity did not significantly change the rooting polarity under the condition of stimulated microgravity, but it increased the competence of explants to IAA treatment. The results presented here provided an experimental system for further investigation of molecular events associated with adventitious root initiation.

[Prokaryote expression and western analysis of BcpLH gene of Chinese cabbage].

BcpLH gene preferentially expressed in folding leaf of Chinese cabbage contains dsRNA-binding domains. The cDNA of BcpLH gene was cloned into a His-fusion expression vector pET-28a (+) and was induced to express in E. coli strain BL21 (DE3). Then, the specific protein was partially purified and the rabbit was immunized to prepare the anti-serum. Meanwhile BcpLH cDNA was cloned into the pMAL-c2 containing the solubizing partner, and then the soluble protein generated. It was demonstrated from Western dot assay that the BcpLH protein was specific. The BcpLH active protein and its anti-serum made it possible to study RNA-binding activity and regulation mechanism in plant development.

Natural antibodies in Microtus fortis react with antigens derived from four stages in the life-cycle of Schistosoma japonicum.

The levels of antibodies which react with the cercarial antigens (CA), schistosomulum stage antigens (SSA), adult-worm antigens (AWA) and soluble egg antigens (SEA) of Schistosoma japonicum were investigated in Microtus fortis and albino mice, using an indirect ELISA. The M. fortis studied fell into three groups: animals caught in the wild; laboratory-bred animals left unchallenged; and laboratory-bred animals that had been challenged with S. japonicum (30 cercariae/animal) 15 days previously. There were also three groups of albino mice: those without infection; those studied 15 days after challenge infection; and those investigated 42 days after infection. The antibodies detected at the highest levels in the laboratory-bred, uninfected voles and in the wild-caught animals were those reacting with SSA, followed, in descending order, by those reacting with AWA, CA and SEA. The levels of natural antibodies to SSA and AWA in these voles were significantly higher than the corresponding levels observed in the uninfected mice and even in the mice infected 15 days previously. The levels of antibodies reacting with CA, SSA, SEA and AWA in the experimentally infected M. fortis were 1.9-, 2.2-, 1.5- and 2.1-fold higher, respectively, than those in the laboratory-bred but uninfected voles. The observations indicate that even uninfected M. fortis produce antibodies which react with S. japonicum, and this presumably results in the natural resistance to infection which has been reported in these rodents.

Potentiation of E7 antisense RNA-induced antitumor immunity by co-delivery of IL-12 gene in HPV16 DNA-positive mouse tumor.

Down-regulation of oncogene expression by antisense-based gene therapy has been extensively studied, and in some cases, therapeutic effects have been demonstrated. We have previously shown that down-regulation of HPV16 E6 and E7 gene expression inhibited HPV DNA-positive C3 mouse tumor growth. Although not all of the tumor cells were transfected by pU6E7AS plasmid, complete tumor regression was achieved if the tumor size was small at the start of therapy in a syngeneic host. This suggests that some other antitumor mechanisms may be involved in addition to the direct down-regulation of HPV16 E7 oncogene expression by the antisense effect of E7AS. In the current study, we demonstrated that E7AS induces tumor cell apoptosis. More importantly, a strong antitumor immune response was elicited in the pU6E7AS-treated and tumor-regressed mice. There was no tumor growth after rechallenging the tumor-regressed mice with 1 million C3 cells. This E7AS-induced antitumor immune response was augmented by co-delivery of mIL-12 cytokine gene. The combination therapy strategy resulted in complete regression of 26 of 28 (93%) tumors. Only 12 of 31 (38%) tumors from the group treated with pU6E7AS alone and 14 of 28 (50%) tumors from the group treated with pCMVmIL-12 alone had completely regressed. Complete regression was also demonstrated in tumors located 1 cm from the treated tumors, which indicates that a systemic antitumor effect was induced by E7AS and mIL-12. Immunohistochemistry demonstrated that a significant amount of CD4+ and CD8+ cells infiltrated into tumors treated with pU6E7AS, pCMVmIL-12 and pU6E7AS+pCMVmIL-12. These data indicate that host immunity is an important factor for antisense-based gene therapy approach which can be further enhanced by combination with cytokine gene therapy.