On the basis of the Guidelines for the diagnosis and treatment of Wilson′s disease 2008, the Chinese guidelines for diagnosis and treatment of Wilson′s disease 2021 has been revised by taking into account the progress of related research in this field both at home and abroad. The original diagnostic criteria and treatment have been supplemented and refined, and the content has been updated until August 2020, mainly involving clinical manifestations, auxiliary examination, diagnosis and differential diagnosis, treatment and prognosis, with the aim of guiding clinicians to standardize the diagnosis and treatment of Chinese patients with Wilson′s disease.

Objective To compare the effects of platelet rich fibrin(PRF)and concentrated growth factors(CGF)using Schwann cells as a pe?ripheral nerve model. Methods A total of 10 healthy volunteers aged 18 to 55 were randomly selected,and 10 mL venous blood was collected un?der aseptic conditions to prepare PRF and CGF. The cells were randomly divided into three groups,control group,PRF group and CGF group. The cell morphology was observed by inverted phase contrast microscope. The cell proliferation was analyzed by MTT assay. The secretion of nerve growth factor in supernatant was detected by enzyme?linked immunosorbent assay. Cell cycle was detected by flow cytometry. Results There was no significant difference of the morphology of cells in each group as observed under inverted phase contrast microscope. MTT results showed that the absorbance values of PRF group and CGF group were significantly higher than those of the control group(P<0.05). The secretion of nerve growth factor in the supernatant were significantly increased(P<0.05). The number of cells in S+G2M phase was significantly increased(P<0.01),but there was no significant difference between PRF group and CGF group. Conclusion Both PRF and CGF can promote proliferation of Schwann cells and increased the amount of nerve growth factor secretion ,but there is no significant difference between PRF and CGF in terms of improving cell proliferation and promoting nerve growth factor secretion.

Objective To probe into methods for postoperatively nursing the patients undergoing pancreaticoduodenectomy combined with vascular allotransplantation. Method The clinical data of postoperative care to 2 patients with pancreatic head carcinoma treated with pancreaticoduodenectomy combined with vascular allotransplantation at the First Affiliated Hospital of Sun Yat-sen University between August 2014 and October 2014 were retrospectively analyzed. Results The two patients lived through the operation, with the operation time of 480 mins, volume of intraoperative blood loss volume of 3000 mL and 600 mL, no complications, hospital stay of 42 days and 27 days. Conclusions The patients undergoing pancreaticoduodenectomy combined with vascular allotransplantation are susceptible to postoperative complications. The nursing care including close observation of disease conditions , drainage tubes care, nutritional care and prevention of complications are critical for the success of operation.

Objective To study the expression of normal and variant ATP7B proteins, in order to further find the mechanism of Wilson disease. Methods Normal ATP7BcDNA/pcDNA3 was made and mutant variants Arg778Leu/pcDNA3, Gln914Ter/pcDNA3 and Thr935Met/pcDNA3 were constructed by using Quik-Change TM Site-directed Mutagenesis Kit in vitro. A good quality rabbit polyclonal antibody against the N-terminal functional domains of ATP7B was produced and purified, being named rabbit anti-human ATP7Bn33-629 polyclonal antibody. Normal and variant expression plasmids constructed above were transfected into Chinese hamster ovary (CHO) cells. After a 36-hour incubation at 37℃, the transfected CHO cells were collected. Expression of normal and variant ATP7B protein were detected and compared by Western blot analysis of cell lysates using ATP7Bn33-629 antibody. Results Expression of ATP7B normal protein in transfected CHO cells was the same as that of ATP7B variant proteins Arg778Leu and Thr935Met.Gln914Ter variant shortened ATP7B protein to 100 kd and increased the level of expression. Conclusion The mechanism under disorder of copper transport caused by the missense mutations should be not related to the level of expression. The increased level of expression caused by Gln914Ter might be associated with the shortened ATP7B protein that needs less time for translation.

Objective To study and report 7 novel mutations and 5 novel polymorphisms of Wilson disease (WD) geneIn combination with the mutations and polymorphisms previously reported, mutation characteristics of WD gene in Chinese were further analysed. Methods Genomic DNA of 60 normal controls and 84 WD patients from 64 families were extracted from peripheral blood leukocytesThe mutations of WD gene (exon1～21) in these subjects were screened by PCR-single strand conformation polymorphism (SSCP) and further confirmed by sequencing. Results 18 different mutations and 17 polymorphisms have been found, in which 7 mutations and 5 polymorphisms are novel, respectivelyOf them, Arg778Leu and Thr935Met reaching a mutation frequency of 37.7% and 10% of in WD chromosomes may be the hotspots of mutation in Chinese population.Ile1148Thr, previously defined as a possible disease-causing mutation may be a polymorphism which has not yet been detected in normal chromosomes. Conclusion In Chinese, WD seems resulting from a few relatively common and a large number more rare mutations

Objective To develop an operational gene diagnosis method for Chinese Facioscapulohumeral muscular dystrophy (FSHD) patients Methods Genomic DNAwas double digested with restriction enzymes EcoRⅠ/HindⅢ and EcoRI/BlnI,respectively The digested fragments were separated on a 0 6% agarose gel After transferred to a Nytran SuperCharge Membrane, the fragmented DNAs were hybridized with the probe p13E 11 The hybridizing fragments were analyzed by the software ImageMaster Total Lab v1 11 and the size of each band was then given Results Only a 4q35 EcoRI+HindIII/P13E 11 fragment larger than 33 kb was detected in each of the controls Two fragments were detected in each of the 33 FSHD patients, one of which was smaller than 33 kb Although there was also presence of two small alleles in the 3 other FSHD cases, either of them turned out to be 10q26 derived owing to its BlnI sensitivity Interestingly, we found a sporadic patient who carried three 4q35 type fragments and, strikingly, two of them were smaller than 33 kb In the analysis of FSHD family members, a 9 year old boy with no clinical signs was found to share the small fragment with his affected father, indicating that he may be a pre symptomatic patient Conclusion The double digestion associated Southern blotting method we developed can be applied to both the diagnosis of FSHD patients and the prediction of pre symptomatic patients Furthermore, by the gene detection using this method, we first got the evidence of translocation between 4q and 10q in Chinese FSHD patients, which may be helpful to the elucidation of the pathogenesis of FSHD

Objective To inestigate the effect of inhaled steroid hormone on the IL-16 in sputum from paitents with asthma, and to explore the mechanism of anti-inflammation of steroid. Method Budesonid inhalant were administered(400?g twice a day) persistent 4 weeks for 12 subjects with mild to moderate asthma, The percentage of eosinophils and IL-16 in induced sputum were determined in every subject .Result After the course , the concentration of IL-16 in sputum was (41 5?5 2)ng/L, significantly lower than that before the therapy which was (58 2?7 2)ng/L,P

Objective To summarize the clinical characteristics of myotonic dystrophy(DM).Methods The clinical data of 24 DM cases were analyzed retrospectively.Results 83.3%(20/24) of the patients obtained the disease during youth and it progressed slowly.79.2%(19/24) of the patients had positive family history.DM was a multisystem disease characterizing by myotonia,weakness and atrophy involved in multiple muscle groups,especially in distal limbs,neck and face.Extensors were more severe than flexors.Spontaneous myotonic discharges and myogenic damages were shown on electromyogram.Pathological examination of muscle biopsies showed increased number of central nuclei,nuclear chains and predominant atrophic typeⅠfibers in 8 cases,muscle fiber necrosis in 7 cases,fibrous structure disorder in 4 cases,sarcoplasmic masses in 3 cases,and serration of sarolemma in 2 cases.Conclusions The clinical characteristics of DM are weakness,atrophy and myotonia.Electromyogram and muscle biopsy are helpful in diagnosis of this disease.

Objective To investigate the perioperative nursing after hemi-hepatectomy and removal of portal vein tumor thrombus(PVTT)for hepatocellular carcinoma(HCC)with PVTT.Methods Fifty-seven HCC patients with PVTT underwent hemi-hepatectomy and removal of PVTT.Careful perioperative nursing was given to each patient.Results All patients successfully lived through surgery.Postoperative complications included 12 cases pleural effusion(21.1%),2 cases refractory ascites(3.5%)and 1 case perioperative death(1.8%).Conclusion Preoperative psychological care,improvement of nutrition and liver function,prevention of upper gastrointestinal bleeding and postoperative observation plus nursing care were of great importance to ensure the success of the surgery.

OBJECTIVE:To provide reference for rational use of antibiotics in the clinic. METHODS:The study quarterly count the dosage of carbapenems,the department distribution of cultured K. pneumonia (271 strains) during Oct. 1st 2013-Sept. 30th 2014,and it’s resistant rates to carbapenems and then investigate the correlation of dosage of carbapenem and drug-resistance of K. pneumoniae. RESULTS:Carbapenem drugs are mainly used in these departments,such as neurosurgery department,ICU and the departments of transplantation and respiratory. Carbapenem-resistant K. pneumonia was also detected in these departments. More-over,with the usage of carbapenem increased by from 64.24% to 360.63%,and the resistant rates of K. pneumonia increased by nearly 10%. CONCLUSIONS:The resistance rates of K. pneumonia is positively related to the amount of carbapenem;the amount control of carbapenems can delay the generation of K. pneumonia.