Electroacupuncture Promotes Functional Recovery after Facial Nerve Injury in Rats by Regulating Autophagy via GDNF and PI3K/mTOR Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To explore the mechanism of electroacupuncture (EA) in promoting recovery of the facial function with the involvement of autophagy, glial cell line-derived neurotrophic factor (GDNF), and phosphatidylinositol-3-kinase (PI3K)/mammalian target of rapamycin (mTOR) signaling pathway.@*METHODS@#Seventy-two male Sprague-Dawley rats were randomly allocated into the control, sham-operated, facial nerve injury (FNI), EA, EA+3-methyladenine (3-MA), and EA+GDNF antagonist groups using a random number table, with 12 rats in each group. An FNI rat model was established with facial nerve crushing method. EA intervention was conducted at Dicang (ST 4), Jiache (ST 6), Yifeng (SJ 17), and Hegu (LI 4) acupoints for 2 weeks. The Simone's 10-Point Scale was utilized to monitor the recovery of facial function. The histopathological evaluation of facial nerves was performed using hematoxylin-eosin (HE) staining. The levels of Beclin-1, light chain 3 (LC3), and P62 were detected by immunohistochemistry (IHC), immunofluorescence, and reverse transcription-polymerase chain reaction, respectively. Additionally, IHC was also used to detect the levels of GDNF, Rai, PI3K, and mTOR.@*RESULTS@#The facial functional scores were significantly increased in the EA group than the FNI group (P<0.05 or P<0.01). HE staining showed nerve axons and myelin sheaths, which were destroyed immediately after the injury, were recovered with EA treatment. The expressions of Beclin-1 and LC3 were significantly elevated and the expression of P62 was markedly reduced in FNI rats (P<0.01); however, EA treatment reversed these abnormal changes (P<0.01). Meanwhile, EA stimulation significantly increased the levels of GDNF, Rai, PI3K, and mTOR (P<0.01). After exogenous administration with autophagy inhibitor 3-MA or GDNF antagonist, the repair effect of EA on facial function was attenuated (P<0.05 or P<0.01).@*CONCLUSIONS@#EA could promote the recovery of facial function and repair the facial nerve damages in a rat model of FNI. EA may exert this neuroreparative effect through mediating the release of GDNF, activating the PI3K/mTOR signaling pathway, and further regulating the autophagy of facial nerves.

Preparation of nanoemulsion spray from Moslae Herba volatile oil and its antibacterial activity / 中国中药杂志

Moslae Herba is a commonly used aromatic Chinese medicinal with volatile oil as the main effective component and exhibits broad-spectrum antibacterial and antiviral effects. However, the irritation and instability of Moslae Herba volatile oil necessitate the preparation into a specific dosage form. In this study, the steam distillation method was employed to extract the Moslae Herba volatile oil. The content of thymol and carvacrol in Moslae Herba volatile oil was determined by HPLC as(0.111 9±0.001 0) and(0.235 4±0.004 7) mg·mL~(-1), respectively. Pseudo-ternary phase diagrams and surfactants compounding were applied in the selection of the optimal excipients(surfactant and cosurfactant). On this basis, a nanoemulsion was prepared from the Moslae Herba volatile oil and then loaded into pressure vessels to get sprays, whose stability and antibacterial activity were evaluated afterward. With clarity, viscosity, smell and body feeling as comprehensive indexes, the optimal formulation of the Moslae Herba volatile oil nanoemulsion was determined as follows: Moslae Herba volatile oil∶peppermint oil∶cremophor EL∶absolute ethanol∶distilled water 7.78∶1.58∶19.26∶6.15∶65.23. The as-prepared nanoemulsion was a light yellow transparent liquid, with Tyndall effect shown under the irradiation of parallel light. It has the pH of 5.50, conductivity of 125.9 μS·cm~(-1), average particle size of 15.45 nm, polydispersity index(PDI) of 0.156, and Zeta potential of-17.9 mV. Under a transmission electron microscope, the Moslae Herba volatile oil nanoemulsion was presented as regular spheres without adhesion and agglomeration. Stability test revealed that the Moslae Herba volatile oil nanoemulsion was stable at 4-55 ℃, which was free from demulsification and stratification within 30 days. After the centrifugation at 12 000 r·min~(-1) for 30 min, there was no stratification either. The nanoemulsion had good inhibitory effects on Escherichia coli, Staphylococcus aureus and resistant S. aureus strains, with the minimum inhibitory concentrations of 0.39, 3.12 and 1.56 mg·mL~(-1), respectively. The above results demonstrated that the nanoemulsion was prepared feasibly and showed stable physical and chemical properties and good antibacterial effects. This study provides a practicable technical solution for the development of anti-epidemic and anti-infection products from Moslae Herba volatile oil.

Clinical and pathological analysis on hydroa vacciniforme-like lymphoproliferative disorder in children / 临床与实验病理学杂志

Purpose To discuss the clinical,histopathological characteristics,diagnosis,differential diagnosis and prognosis of hydroa vacciniforme-like lymphoproliferative disorder in children.Methods 6 cases of hydroa vacciniforme-like lymphoproliferative disorder were analyzed by molecular,histopathological and immunohistochemical testing.Clinical and follow-up information was obtained.The published relevant literatures were reviewed.Results 4 cases were boys,2 case were girls.All the patients presented with erythema and blisters with fever for 1 month to 4 years.Histopathologic examination showed an mild or moderate atypical lymphocytic infiltrate with angiotropism and angiocentricity,and scattered or dense lymphoid infiltration throughout the dermis and subcutaneous tissue.Blisters or necrosis could be seen in the epidermis.The atypical lymphocytes were positive for CD2,CD3,CDS,CD7,CD43,TIA-1,CD4 or CD8,and negative for CD20,Pax-5.Only one case showed positive stain for CD56.The average positive rate of Ki67 in tumor cells was 42.3％.Tumor cells positive for EBV encoded RNA (EBER) were detected in cutaneous infiltrates in 5 cases.Gene rearrangement of TCR was detected in 2 cases.5 patients were available for follow-up examination and 1 patient was dead.Conclusion Hydroa vacciniforme-like lymphoproliferative disorder is a rare disease mainly occuring in children.Chronic active EBV infection has been associated with this disease.It may be a spectrum in terms of its clinical course,and may be benign,borderline and malignant.Pathological diagnosis should be closely combined with clinical data.

Silencing endothelin-3 expression attenuates the malignant behaviors of human melanoma cells by regulating SPARC levels / 华中科技大学学报（医学）（英德文版）

Endothelin-3 (ET-3) is aberrantly expressed in both metastatic melanoma tissues and cultured melanoma cells. Our previous work showed that ET-3 could promote survival of metastatic melanoma cells via its altered expression. In this study, we investigated the mechanisms responsible for these gene-induced phenotypes in melanoma cells. An ET-3 gene sequence-specific shRNA vector pLVTHM-ET3-RNAi was constructed and transfected into human malignant melanoma cells A375 and MMRU, and the resultant molecular events and cellular changes were examined. As compared with the empty-vector group, cell proliferation was slowed down, and the growth inhibition rates were 38.9% in A375 cells and 38.4% in MMRU cells after transfection. In addition, cell invasion capability was also inhibited, with a reduction of 62.2% in A375 cells and 54.3% in MMRU cells. The percentage of apoptotic cells was found to increase. Meanwhile, in both cell lines, secreted protein acidic and rich in cysteine (SPARC) levels were down-regulated together with inhibition of its upstream signaling molecule, NF-κB. Thus, the current results suggested that down-regulated expression of ET3 attenuates the malignant behaviors of human melanoma cells partially by decreasing the expression of SPARC and NF-κB.

Silencing endothelin-3 expression attenuates the malignant behaviors of human melanoma cells by regulating SPARC levels / 华中科技大学学报（医学）（英德文版）

Endothelin-3 (ET-3) is aberrantly expressed in both metastatic melanoma tissues and cultured melanoma cells. Our previous work showed that ET-3 could promote survival of metastatic melanoma cells via its altered expression. In this study, we investigated the mechanisms responsible for these gene-induced phenotypes in melanoma cells. An ET-3 gene sequence-specific shRNA vector pLVTHM-ET3-RNAi was constructed and transfected into human malignant melanoma cells A375 and MMRU, and the resultant molecular events and cellular changes were examined. As compared with the empty-vector group, cell proliferation was slowed down, and the growth inhibition rates were 38.9% in A375 cells and 38.4% in MMRU cells after transfection. In addition, cell invasion capability was also inhibited, with a reduction of 62.2% in A375 cells and 54.3% in MMRU cells. The percentage of apoptotic cells was found to increase. Meanwhile, in both cell lines, secreted protein acidic and rich in cysteine (SPARC) levels were down-regulated together with inhibition of its upstream signaling molecule, NF-κB. Thus, the current results suggested that down-regulated expression of ET3 attenuates the malignant behaviors of human melanoma cells partially by decreasing the expression of SPARC and NF-κB.

Effect of alendronate sodium on torque-out testing on implant-bone interfaces in estrogen-deficient rabbits with alendronate systemic administration / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the influence of systemic application of Alendronate sodium, a bone resorption inhibitor, on the osseointegration of implant-bone interface in estrogen-deficient rabbits through mechanical assessment.</p><p><b>METHODS</b>27 five-month-old Japanese white female rabbits were randomly divided into three groups (9 rabbits each group). An ovariectomy group (OVX), an ovariectomy and Alendronate sodium group (ALN) and a shamed-operated group (S). 12 weeks after operation, implants were installed into bilateral distal femurs and proximal tibias in each group. Alendronate sodium was administrated by intraperitoneal injection in ALN group; meanwhile equivalent of normal saline was administrated by intraperitoneal injection in OVX group and S group. Bone mineral density was measured right after the implant operation and also in 4, 8, 12 weeks. Torque-out values were measured in 4, 8, 12 weeks after animal sacrifice.</p><p><b>RESULTS</b>Bone mineral density of tibias in ALN group was closed to S group and was significantly different from OVX group (P < 0.05) after 8 weeks. While after 12 weeks, the bone mineral density of tibias and femurs in ALN group was both closed to S group and was significantly different from OVX group (P < 0.05). The torque-out values of tibias in ALN group were closed to S group and were significantly different from OVX group (P < 0.05) after 8 weeks. After 12 weeks, the torque-out values of tibias and femurs in ALN group were both closed to S group and were significantly different from OVX group (P < 0.05).</p><p><b>CONCLUSION</b>Systemic application of Alendronate sodium in osteoporosis rabbits can improve the bone-implant osseointegration significantly.</p>

Retrospective analysis of 125 cases of pregnancies complicated by rheumatic heart disease / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the impact of rheumatic heart disease on pregnancy.</p><p><b>METHODS</b>A retrospective study was conducted in 125 pregnant women with rheumatic heart disease. These cases were divided into operation (group A) and non-operation (group B), and the cardiac function, pregnancy complications, gestational weeks, delivery modes and outcomes of pregnancy were compared.</p><p><b>RESULTS</b>The cardiac function was significantly improved and pregnancy complications reduced after the cardiac operation with extended gestational weeks, showing significant differences between the two groups (P<0.05). One perinatal fetal death without maternal death occurred in group A, as compared with two maternal and two perinatal deaths in group B. Cesarean section was the primary delivery mode in these cases.</p><p><b>CONCLUSION</b>Surgical interventions can improve the cardiac function, reduce the pregnancy complications and improve the pregnancy outcomes in pregnant women with rheumatic heart disease. Cesarean section is the primary choice for pregnant women with prosthetic heart valve replacement.</p>

Isolation and identification of channel catfish (Ictalurus punctatus) hemorrhage reovirus / 病毒学报

By using cell culture and virus infection methods, a new reovirus had been isolated from channel catfish (Ictalurus punctatus) suffered with severe hemorrhage and had been identified as channel catfish reovirus (CCRV) after artificial infection in fish, electron microscopy observation, physical-chemical tests, genomic SDS-PAGE analysis and sequencing. In artificial infection test, the typical symptoms of channel catfish hemorrhage as naturally occurred could be reproduced. The isolated virus could cause typical cytopathic effect in CCO and CCK cell lines. Electron microscopy observation of ultra-thin section samples of CCRV infected CCO and CCK cells revealed that the virus replicated in cytoplasm, arrayed in crystalline, and had a non-enveloped double capsid with a diameter of 60-70 nm. Frozen-thawed, 56 degrees C 1 h, chloroform and ether had no significant effects on CCRV titer, 65 degrees C 1 h could significantly inactivated the viral infectivity. The CCRV genome SDS-PAGE analysis and nuclease sensitivity test showed that the virus genome was the same as that of viruses in Aquareovirudae and consisted of 11 segments of dsRNA assigned into three classes L1, L2, L3; M1, M2, M3 and S1, S2, S3, S4, S5 with a range of size from 0.9 to 4.4 kb. The Cloning and sequencing of the CCRV S4 segment indicated the nucleic acid number of CCRV S4 was 909 bp in length, which was exactly the same as that of GCRV S4 (AF403396) and GSRV S4 (AF403407) segments. The BLAST of CCRV S4 sequence in NCBI GenBank showed that it had a 99% and 90% similarity in sequence to the GCRV S4 and GSRV S4 segments, respectively.

Highly efficient inhibition on replication of grass carp reovirus mediated by chemically synthesized small interfering RNAs / 病毒学报

Two short interfering RNAs (siRNA-RdRp1286, siRNA-RdRp1441) and one short interfering RNA (siRNA-OCP117), targeted to the RNA dependent RNA polymerase (RdRp) gene and outer capsid protein (OCP) gene of Grass carp reovirus (GCRV) respectively, were chemically synthesized and transfected into the CIK cells by lipofectamine 2000. 6 hours after transfection, the transfected CIK cells were challenged with GCRV. The culture media were collected at 48h post challenge and the virus was titrated in microculture system to evaluate the inhibition effect on GCRV replication mediated by siRNAs. Referring to the mRNA level of housekeeping gene beta-actin, RT-PCR was applied to detect the level of GCRV mRNA in transfected and challenged CIK cells. The results showed that the viral titer (lgTCID50/0. 1mL) in siRNA-RdRp1286, siRNA-RdRp1441 and siRNA-OCP117 transfected CIK cells were 4.41 +/- 0.16, 3.83 +/- 0.44 and 1.94 +/- 0.42 respectively, which were significantly lower than that in virus infection positive control (7.92 +/- 0.52) (P < 0.01). No significant change in viral titer was observed in the group transfected with siRNA negative control after challenged with GCRV (7.50 +/- 0.17, P > 0.05). Compared with the mRNA transcriptional level of beta-actin gene in virus infection positive control, the mRNA levels of GCRV in siRNA-RdRpl 286, siRNA-RdRp1 441 and siRNA-OCP117 transfected CIK cells were reduced significantly and the inhibition rate reached to (82.08 +/- 2.15)%, (89.19 +/- 1.14).% and (92.62 +/- 0.17)%, respectively. The mRNA level of GCRV in the siRNA negative control group had no noticeable change (P > 0 05).

Study on the protective effect of recombinant human N-terminal lipopolysaccharide binding protein in mice challenged with LPS / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of recombinant human N-terminal lipopolysaccharide binding protein in mice challenged with LPS.</p><p><b>METHODS</b>Seventy male Kunming mice were randomly divided into 3 groups, i.e. LPS challenge (Injection of LPS into abdominal cavity, n = 21); tLBP protection (Injection of LPS and tLBP into abdominal cavity, n = 21) and control (Injection of normal saline into abdominal cavity, n = 8) groups. The blood samples and tissue samples of the liver and lungs were harvested on 15 and 30 minutes and 1, 3, 6, 12 and 24 hours after the injection. The serum contents of ALT and TNF-alpha were determined by biochemical velocity analysis and RIA method, respectively. The pathomorphological changes in the liver and pulmonary tissue were examined under light microscope (LM). The mortality rate of ten mice each was observed within 24 hours after the injection of tLBP + 400 ng LPS or 400ng LPS.</p><p><b>RESULTS</b>The ALT content of tLBP group reached the peak level at 12 post-injection hour (PIH) (41.00 +/- 4.58), but it was significantly lower than that in LPS group in which it peaked at 6PIH (99.50 +/- 62.63) (P < 0.01). The TNF-alpha content in tLBP and LPS group was lower than that in LPS group, and both reached the peak level at 3 PIH (35.96 +/- 7.33). Compared with those in LPS, injury to hepatocytes in tLBP group was obviously milder without scattered necrosis. The pulmonary congestion in tLBP group was abated, and the inflammatory exudation in the alveoli was evidently less than that in LPS group. There were 9 out of 10 mice died in the LPS challenge group, while only 3 out of 10 mice died during 24 hours after LPS injection in tLBP protection group.</p><p><b>CONCLUSION</b>Preliminary results indicated that recombinant human tLBP might possess biological activity with a potential protection effect in LPS challenged mice.</p>

Granulocyte colony-stimulating factor gene polymorphism in patients with systemic lupus erythematosus in Northern Chinese / 中华风湿病学杂志

Objective To analyze the polymorphism(s) or mutation(s) of the granulocyte colony-stim- ulating factor gene (G-CSF,Csf3) and its possible association with the susceptibility to systemic lupus erythe- matosus (SLE).Methods Polymorphism screening was carried out by the polymerase chain reaction-single strand conformation polymorphism method (PCR-SSCP),using DNA from 204 Northern Chinese patients with SLE,459 ethnically matched healthy control,78 patients with other autoimmune diseases.Results It was i- dentified that 4 polymorphisms at position 1869,1205,1931,and 394.The nucleotide sequences of the sample that showed different SSCP patterns and different distribution between SLE and healthy controls were deter- mined by direct sequencing.Only the distribution of the fifth extron of Csf3 (1931) between SLE and controls was different.The frequency of A/G genotype (60.5%) and A allele (53.2%) at position 1931 in patients with SLE was significantly higher than that in healthy controls (51.6% and 45.2%,respectively),whereas,the fre- quency of G/G genotype (16.2%) and G allele (46.8%) in patients with SLE was lower than that in healthy controls (28.9% and 54.8% respectively).Conclusion The results suggest that Csf3 (1931) is significantly associated with the susceptibility to SLE in Northern Chinese patients .Further population and functional stud- ies will be followed to establish Csf3 (1931) as one of the susceptible genes for SLE.

Modified tubo-uterine implantations for proximal tubal occlusive infertility after femal sterilization with mucflago phenol / 中华妇产科杂志

Objective To explore the effects of modified tubo-uterine implantations performed on women with proximal tubal occlusive infertility after femal sterilization with mucilago phenol.Methods Two hundred and eight infertile women who were admitted to the Second Affiliated Hospital of Sun Yat-sen University between 1986 and 2004 were included.They all accepted modified tubo-uterine implantation after occlusion of fallopian tubes with mucilago phenol.Results It was found that the occlusions were all located in the interstitial portion or isthmic portion of the fallopian tubes.Different degrees of pelvic adhesions were found in 65 cases.Fifty-seven cases were slightly adhesive,seven cases were of moderate degree and one case was severe.One hundred and ninety-nine cases were followed up after operations(95.7%).One hundred and ninety-three women accepted hydrotubation in the following month just after the operation and 185 women were found to be unobstructed(95.8%).One hundred and forty-three women became pregnant, the pregnant rate being 71.9%(143/199).One hundred and twenty-five women had term deliveries (87.4%),three women were in early pregnancy and two in midtrimester pregnancy.Eleven women had spontaneous abortion(7.7%).Two women had tubal pregnancy(1.0%).None of the 199 cases had any signs of endometriosis.Conelusions Modified tubo-uterine implantations are quite effective for proximal tubal occlusive infertility.It may be a favorable method for such kind of tubal occlusions.

CONSTRUCTION AND IDENTIFICATION OF HCMV cDNA EXPRESSING LIBRARY AND SCREENING OF pp65 POSITIVE CLONES / 微生物学通报

In order to provide effective tool for further studying of the function of HCMV genome, developing of molecular vaccine and diagnostic reagents. Extraction of HCMV mRNA from HF cell infected by HCMV AD169 strain for 96h was reverse transcripted into cDNA, then was cloned into EcoR I-digested lambda gt11. HCMV AD169 strain cDNA expressing library has been constructed after packaging. The volume and the recombination rate of the prime cDNA expressing libraries was 3.6?10 6 and 96%, 168 positive clones of HCMV were screened by immune blotting with anti-HCMV mouse convalescent sera, 34 positive clones were obtained by dot nucleic acid hybridization with DIG-labled HCMV pp65 gene probe. 2 positive clones were amplified by HCMV pp65 all length primer. The PCR product has been tested by southern-blotting.The PCR product was sequenced and was taken as homology comparison by DNASIS software,and the homology is 98%.To lay the foundation of furher cloning,expressing the pp65 gene,further studying of the function of the pp65 prodct.