Objective To package the recombination adeno-associated virus with human U6 promoter and c-met short hairpin in order to establish foundation for research on inhibiting the expression of c-Met and cancer gene therapy.Methods c-Met short hairpin was synthesized and linked to the down stream of U6 promoter by PCR.Adeno-associated viral vectors pSNAVUshMet(1,2) were constructed and transfected into BHK cells.Positive cell clones were selected by G418.Adeno-associated virus with U6shMet were packaged by adding rHSV1-repcap.The virus purity was analysed by SDS-PAGE and titer was assayed by dot blot.Results Two fragments(U6shMet1 and U6shMet2) with c-Met short hairpin were obtained and two recombination adeno-associated virus(rAAVUshMet1 and rAAVUshMet2) with U6shMet were packaged successfully.SDS-PAGE analysis showed that the purified virus appeared clear characterized bands.The virus titer was 4?1012mg?L-1.Conclusion The recombination adeno-associated virus(rAAVUshMet1 and rAAVUshMet2) with U6shMet are constructed successfully.The virus have high titer and good purity and could act as the effective vehicle of inhibiting the expression of c-Met and cancer gene therapy.

Objective To investigate the inhibitory effect of downregulation of hepatitis B virus (HBV) core gene (HBcAg) expression by RNA interference and magnetic nanoparticles on both HBV DNA replication and expression in vitro. Methods HepG2 2.2.15 cells were transfected with U6 promoter plasmids coding for small interfering RNA (siRNA) targeting HBV core gene using magnetic nanoparticles. RT-PCR and Western blot were used to assess the mRNA and protein expression HBV core antigen. Real-time PCR was used to evaluate the suppression efficiency of HBV-DNA replication and expression; and radioimmunoassay was used for HBV surface antigen (HBsAg), core antigen (HBcAg), and e antigen (HBeAg) detection. Results We successfully constructed nanoparticles with siRNA plasmid targeting HBV core antigen; HBcAg mRNA and HBV core antigen protein levels were significantly reduced in the transfected cells. HBV-DNA downregulation was estimated at 4-5 logs and the HBsAg and HBeAg levels were also reduced compared with the controls. Conclusion Downregulation of HBV core gene using RNAi technology and magnetic nanoparticles can potentially be used as a therapeutic strategy for Hepatitis B.

Objective To evaluate the indications,techniques and effects of high speed drill assisted cervical pedicle screw-rod system in treatment of cervical interlocking dislocation.Methods A retrospective review was made on data of 13 cases of cervical interlocking dislocation undergone pedicle screw-rod fixation and reduction with assistance of high speed drills from December 2006 to July 2011.Dislocation localized in C3/4 (n =3),C4/5 (n =5),C5/6 (n =4) and C6/7 (n =1) respectively.Causes for injury contained cervical hyperextension due to traffic accidents (n =7) and a fall from the height (n =6).Two remained neurological intact,seven were combined with syndrome of central canal damage,and four had paraplegia.All the 13 cases received posterior pedicle screw fixation and reduction within 7 days after injury.Pedicle screws were implanted on adjacent segment of cervical spine with unlocking the interlocked small joints by a distraction device.Results All 13 cases obtained good reduction,including eight with complete anatomic reduction and five with 1-2 mm of vertebral slight slippage.Fifty-two screws were inserted and 49 pedicle screws were put in place except for the excursion in three pedicle screws on the postoperative radiographic and CT scans.There was no injury of spinal cord,nerve root or vertebral artery during operation.The patients showed significant improvement in sensory and motor function of the upper extremities in the follow-up of 12-36 months.One case had screw breakage at postoperative three months with no nerve symptoms.Conclusion High speed drill assisted pedicle screw-rod system is suitable for treatment of cervical interlocking dislocation,for its reduction process accords with the biomechanics and allows decompression therapy.

Objective To compare the clinical effects of laparoscopic Nissen fundoplication (LNF)combined with highly selective vagotomy(HSV)(LNFHSV)in treatment of gastroesophageal reflux disease(GERD),and to provide reference for the clinical application of LNFHSV.Methods 22 patients with a history of GERD unrelieved by medication and underwent LNFHSV were selected.At the same time ,36 patients with GERD underwent LNF were used as control.The mean operation time,hospital stay,incidence of main operative complications,HSS complete remission and Demeester scores of the patients in two groups were retrospectively analyzed.Results The Demeester scores,hospital stay, incidence of main operative complications of the patients in two groups before operation had no significant differences(P>0.05).The mean operation time in LNFHSV group (90 min±35 min) was longer than that in LNF group(65 min± 21 min).The Demeester scores of the patients in two groups after operation had significant difference(P<0.05).The HSS complete remission rate of the patients in LNFHSV group was 91.1%,the part remission rate was 8.9%,and no effectiveness was 0;they were 83.3%,13.8%,and 2.8% in LNF group;there were significant differences between two groups(P<0.05).Conclusion LNFHSV has better effectiveness in controlling GERD than LNF procedure.

Objective To improve the diagnostic and therapeutic ability of Brucellosis by analyzing the epidemic and clinical characteristics.Methods A retrospective analysis was done on the data of Brucellosis patients treated in our hospital from 2007 to 2016.Results Since the first case was diagnosed in 2012,27 patients [19 male and 8 female,mean age (44.4 ± 16.9) years] were confirmed by clinical manifestations and positive bacterial cultures results.The annual number of cases from 2012 to 2016 was 1,1,6,4 and 15.Among them,10 cases (37.0％) had a history of close contact with goat,7 cases (25.9％)with raw mutton,1 case (3.7％) with raw beef and 1 case (3.7％) with suspicious laboratory contamination while 8 cases (29.6％) had no evident risk factors for Brucellosis.The common clinical manifestation included fever (81.5％),lumbago/joint pain (55.6％),fatigue (33.3％) and hyperhidrosis (22.2％).The white blood cell count was normal among 20 cases (74.1％) while 6 cases (22.2％) with leukopenia.Mild to moderate anemia in 20 cases (74.1％) and decreased platelet number in 4 case (14.8％).The percentage of elevated alanine aminotransferase,aspartate aminotransferase and lactate dehydrogenase was 32.0％,48.0％ and 100％,respectively.Decreased albumin level was found in 23 cases (92.0％).The percentage of elevated erythrocyte sedimentation rate (ESR),C-reactive protein and serum ferritin 75.0％,82.3％ and 77.8％,respectively,while 12 cases (85.7％) with procalcitonin level below 0.5 ng/L.According to follow-up for at least half year,all the cases were cured by active medical management.Conclusions The number of Brucellosis cases is rapidly increasing in our hospital.It's of great significance to know the epidemic and clinical characteristics of Brucellosis.

Objective To systematically evaluate the efficacy and safety of plum-blossom needle therapy for vitiligo by using the systematic review and meta analysis.Methods The randomized controlled trials(RCT)on plum-blossom needle for treating vitiligo were systematically retrieved from the databases of the PubMed,China Biological Medicine Database,CNKI,Wanfang Data and VIP database from the database estab-lishment to June 2,2022.The literatures were screened according to the inclusion and exclusion criteria.The finally included literatures conducted the data extraction.The RevMan 5.4 software was used for conducting the data analysis.The methodological quality evaluation on the included trials was performed by the ROB tool.The GRADE method was used to assess the evidence level.Results A total of 7 trials involving 469 pa-tients were finally included.The meta analysis results showed that the plum-blossom needle combined with other therapies(including laser or ultraviolet irradiation,tacrolimus ointment,compound Kaliziran tincture)was superior to the other therapies alone in the aspects of improving vitiligo skin lesion including reducing the of white spot skin lesion area(MD=-1.11,95％CI:-1.92 to-0.30,P=0.007),increasing the repigment-ation rate of vitiliginous lesions(MD=18.09,95％CI:1.55 to 34.63,P=0.030)and enhancing the pigment deposition in vitiligo lesions(MD=0.92,95％CI:0.32 to 1.52,P=0.003),and improving the patients'quali-ty of life(MD=-7.48,95％CI:-8.04 to-6.92,P＜0.001),and the differences were statistically signifi-cant.In terms of safety,there was no statistically significant difference in adverse events between plum blos-som acupuncture combined with other therapies and other therapies alone(RR=1.20,95％CI:0.77 to 1.84,P=0.420).Conclusion Low or very low evidence shows that plum-blossom needle combined with other therapies for treating vitiligo may enhance the effect in the aspects of improving the white spot skin lesions and quality of life in the patients with vitiligo,moreover which is relatively safe.

Subcortical vascular mild cognitive impairment (svMCI) is a common prodromal stage of vascular dementia. Although mounting evidence has suggested abnormalities in several single brain network metrics, few studies have explored the consistency between functional and structural connectivity networks in svMCI. Here, we constructed such networks using resting-state fMRI for functional connectivity and diffusion tensor imaging for structural connectivity in 30 patients with svMCI and 30 normal controls. The functional networks were then parcellated into topological modules, corresponding to several well-defined functional domains. The coupling between the functional and structural networks was finally estimated and compared at the multiscale network level (whole brain and modular level). We found no significant intergroup differences in the functional–structural coupling within the whole brain; however, there was significantly increased functional–structural coupling within the dorsal attention module and decreased functional–structural coupling within the ventral attention module in the svMCI group. In addition, the svMCI patients demonstrated decreased intramodular connectivity strength in the visual, somatomotor, and dorsal attention modules as well as decreased intermodular connectivity strength between several modules in the functional network, mainly linking the visual, somatomotor, dorsal attention, ventral attention, and frontoparietal control modules. There was no significant correlation between the altered module-level functional–structural coupling and cognitive performance in patients with svMCI. These findings demonstrate for the first time that svMCI is reflected in a selective aberrant topological organization in multiscale brain networks and may improve our understanding of the pathophysiological mechanisms underlying svMCI.

OBJECTIVE: To determine features of the clinical manifestation in patients with primary biliary cirrhosis (PBC), and to provide a scientific basis for diagnosis of PBC.
 METHODS: A total of 102 patients with PBC treated in the Second Xiangya Hospital, Central South University, from January 2013 to January 2015 were retrospectively analyzed. The patients' general condition, clinical manifestations, serum biochemical and immunological parameters were detected.
 RESULTS: Of the 102 PBC patients, 91 (89.21%) patients were female. The main symptoms in these patients were fatigue, poor appetite, dry mouth, nausea, vomiting, pruritus, stomachache, and abdominal distension. The major signs were jaundice, splenomegaly, hepatomegaly, edema, and ascites. The main features of serum biochemical parameters in these patients included the increase of alkaline phosphatase and gamma glutamyltranspeptidase (GGT), especially the GGT. The anti-mitochondrial antibodies-M2 (AMA-M2) in 81 and 21 patients was positive and negative, respectively. The differences between the AMA-MA positive and negative groups were not statistically significant (P>0.05). According to clinical manifestation, 102 patients were classified into 2 groups: A non-cirrhosis group (n=56) and a cirrhosis group (n=46). The positive rates in these 2 groups, such as ANA, AMA-M2, anti-gp210, anti-Sp100, anti-Ro52, anti-PML, were 54.35%, 89.13%, 41.30%, 13.04%, 43.38% and 10.87% vs 57.14%, 71.43%, 42.86%, 12.5%, 51.79% and 3.71%, respectively, with no significant difference between them (P>0.05). However, there was significant difference in the positive rate of anti-3E-EPO between the above 2 groups (86.78% vs 58.93%, P<0.05). The positive rates of AMA-M2 and anti-3E-EPO in 30 patients diagnosed by hepatic histopathological examination were higher than those of other antibodies.
 CONCLUSION PBC mainly affects middle-aged women, and its clinical manifestation is various. The autoantibody tests play an important role in diagnosis of PBC. Checking for AMA-A2 and anti-3E-BPO can improve the positive rate of PBC. Liver histopathological examination may provide useful information on disease severity, which can determine the histological stage when the patient's serum autoantibodies are negative.
Alkaline Phosphatase ; metabolism ; Autoantibodies ; blood ; Female ; Humans ; Liver Cirrhosis, Biliary ; diagnosis ; pathology ; Male ; Mitochondria ; Retrospective Studies ; gamma-Glutamyltransferase ; metabolism