Objective To prospectively evaluate the risk factors for the decline of residual renal function (RRF) in new peritoneal dialysis (PD) patients.Methods A total of 84 new PD patients in our PD center were included in this study.Clinical comprehensive assessment were made,and regression models was established to analyze the relationship between the decline of RRF and clinical indicators,which included the rate of peritonitis,systolic pressure,diastolic pressure,urine volume,24 h urinary protein,serum albumin,C-reactive protein(CRP),history of diabetes mellitus,and the use of angiotensin converting enzyme inhibitor (ACEI) or angiotensin receptor blockers (ARB) drugs,cardiac functional grading,sodium and water retention and biochemical indicators.The primary outcome was defined as two consecutive urine volume ≤ 100 ml/24 h.Results The mean follow-up time was (11.7± 1.1) months,primary outcome occurred in 20 patients,accounting for 23.8％,and their average period progressed to the primary outcome was (10.5±2.0) months.The 20 patients had higher ultrafiltration volume [(551.6±328.2) ml vs (294.1±288.0) ml,P=0.001],higher systolic blood pressure [(145.2±16.5) mmHg vs (136.0±13.8) mmHg,P=0.016],worse cardiac functional grading [(1.7±0.8) vs (1.3±0.4),P=0.000],more serious water-sodium retention [(1.0±0.7) vs (0.6±0.5),P=0.012],higher peritonitis rates (35.0％ vs 4.7％,P=0.000),lower Kt/V [(1.7 ± 0.4) vs (2.0 ± 0.3),P=0.003],lower hemoglobin levels [(89.0± 14.9) g/L vs (99.5± 17.8) g/L,P=0.020],higher C-reactive protein levels [(19.4±34.4) mg/L vs(8.7±12.6) mg/L,P=0.017],higher Scr levels [(1 004.6±291.1) μmol/L vs (753.1±254.3) μ mol/L,P=0.000],lower serum calcium levels[(1.86±0.1) mmol/UL vs (2.02±0.2) mmol/L,P=0.000],higher serum phosphorus [(2.1±0.6) mmol/L vs (1.6±0.4) mmol/L,P=0.001] and higher calcium phosphorus product [(3.8±1.1) mmol2/L2 vs (3.1±0.8) mmol2/L2,P=0.010] as compared with those of the patients without the primary outcome.Based on the results of multivariable Cox regression analysis,ultrafiltration volume,cardiac functional grading,peritonitis,Kt/V and serum phosphorus level contributed to the decline of RRF significantly.Conclusion The higher Kt/V in PD patients plays a protective role,the higher ultrafiltration volume,the worse heart function,the more peritonitis rate and higher serum phosphorus predict more rapid declination of RRF.

[Objective] To sum up teacher He Lingzhi’s clinical experience and academic ideas on diagnosing and treating IgA nephrosis. [Method] From casa morbi, mechanism and clinical treatment based on differentiation of signs, it expounds teacher He Lingzhi ’s clinical experience and academic points, exampled with a case. [Result] The disease has main mechanism of Qi deficiency, teacher He Lingzhi divides the IgA nephrosis into types of Qi deficiency of lung and spleen, spleen and kidney, both deficiency of lung and kidney, Yang deficiency of spleen and kidney, Yin deficiency of liver and kidney, both deficiency of Qi and Yin, Yin and Yang, thus forms the treating rules of tonifying Qi to reinforce spleen, tonifying lung to nourish kidney, tonifying liver and kidney, Qi and Yin, Yin and Yang. The case here was relieved the nephrosis symptoms with the treatment. [Conclusion] Teacher He Lingzhi has rich clinical experience in treating IgA nephrosis with differentiation of signs, with definite cure effect.

Abdominal withdrawal reflex (AWR) test, which evaluates the behavioral response of animals through expanding the intestinal tract, is the most widely used method to assess animal's visceral sensitivity in recent years.However, as AWR experimental operation involves many factors, it''s hard to find a commonly recognized operating procedure.Through review of literature of AWR experimental measuring methods, this article summarizes the basic steps of AWR operation.Meanwhile, it scrutinizes the differences of various measuring methods in relevant factors, including expansion method and airbag method, with special operating suggestions to them.Hopefully, this article will provide an option to future researchers who will use this method to evaluate animal''s visceral sensitivity, and provide a necessary foundation for the standardization of the experimental operation.

Objective To optimize the preparation of air bag materials for abdominal withdrawal reflex test.Methods To measure the diameters of different pressue-air bags by digital vernier caliper and X ray imaging.Using the pain threshold (3 points) as a criterion to test the pressure values of different air bags in the rats with corresponding visceral pain sensitivity.Results With the increasing pressure, the changes of the diameter of round latex balloon were uniform and gentle, and could meet the pressure range used in the experiment (20~80 mmHg).The threshold of visceral pain in rats was near the nociceptive stimulus value,and would not cause irreversible damage to the rat intestinal tract.Conclusions Air bag made from round latrex ballon is an ideal tool to be used in abdominal withdrawal reflex test in rats.

Objective To investigate the effect of lipid emulsion on mitochondrial energy metabolism during bupivacaine-induced myocardiotoxicity in rats.Methods H9c2 cells were inoculated in 96-well plates at a density of 1 × 105cells/ml, and were randomly divided into 4 groups (n =18 each) using a random number table: control group (group C) , bupivacaine group (group B) , lipid emulsion group (group LE) , and bupivacaine + lipid emulsion group (group B + LE).The cells were incubated in the normal culture medium in group C.In group LE, the cells were incubated in the culture medium containing 1％ lipid emulsion.In group B, the cells were incubated in the culture medium containing 1 mmol/L bupivacaine.In group B + LE, the cells were incubated in the culture medium containing 1 mmol/L bupivacaine and 1％ lipid emulsion.After 24 h of incubation, the contents of ATP, ADP, and AMP were measured by high-performance liquid chromatography, and apoptotic rate was calculated by Hoechst33342/ PI staining.Results Compared with group C, the contents of ATP, ADP and AMP were significantly decreased, and apoptotic rate was increased in group B (P＜0.05), and the contents of ATP and ADP in group LE and ATP content in group B + LE were increased, and no significant changes were found in apoptotic rate in LE and B+LE groups (P＞0.05).Compared with group C, the contents of ATP, ADP and AMP were significantly increased, and apoptotic rate was decreased in LE and B+LE groups (P＜ 0.05).Compared with group LE, the contents of ATP, ADP and AMP were significantly decreased (P＜ 0.05), and no significant change was found in apoptotic rate in group B+LE (P＞0.05).Conclusion The mechanism by which lipid emulsion reduces bupivacaine-induced myocardiotoxicity may be associated with improved mitochondrial energy metabolism in rats.

Objective: To study the difference between the acid resistance of Streptococcus mulans Ingbritt C international standard strain and the acid resistance of LuxS mutant strain. Methods: Solutions of Streptococcus mulans standard strain and LuxS mutant strain with same density were prepared and cultured at pH 3. 5 to 7. 0 BH1 liquid for same period. Terminal growth situation was compared. After being acidized in pH 5.5 BHI liquid, the two strains were cultured at pH 3.0 BHI liquid. The acid tolerance responses of the two strains were compared. Results; (DAt pH 6.0 to 7. 0, the difference of growth between Streptococcus mulans standard strain and LuxS mutant strain was not significant at the same pH value, and the differences of bacterial growth situation under three different pH values were not significant. (1)At pH 4.5 to 5.5, the difference of growth between the two strains was significant. (2)At pH 3.0,the survival rate of LuxS mutant strain(0.006 5% )was significantly lower than the standard strain (0.078% ). (3)At pH 5.5, the survival rate of LuxS mutant strain(0.747% ) was lower than the standard strain(8.65% )by about 10 times after the pre-acidification. Conclusion; (4)At sub-lethal pH value, there is significant difference of aciduricity between Streptococcus mu-tans standard strain and LuxS mutant strain. The acid resistance of standard strain is stronger than that of LuxS mutant strain. The two strains both display the capability of acid tolerance responses. LuxS mutant strain is more sensitive to acid inactivation, but the capability of acid tolerance responses still exists.

Objective Mikulicz's disease (MD) was considered to be a subtype of Sj(o)gren's syndrome (SS) due to the clinical and histological similarities between them.Evidence had shown that there were differences between MD and typical SS.The purpose of this study was to investigate the correlation between MD and SS,by means of analyzing the expression of IgG4 in salivary glands and the clinical characteristics of patients who were previously considered as SS.Methods The paraffin sections of salivary glands from SS patients were stained with monoclonal antibodies to IgG4 and CD38.Patients were divided into two groups based on the pathological results.Analysis of the symptoms,the signs and the laboratory results were carried out in these patients.The difference in laboratory parameters and histopathological gradings in the two groups was analyzed.Normal and abnormal distributed data comparison was conducted using random independent samples t-test and rank sum test respectively.Two-sample rates were compared with Chi-square test.Results Based on immunohistochemistry of IgG4 distribution,the 58 patients with SS were divided into two groups:IgG4 related (9 cases) and non-IgG4 related (49 cases).Histopat-hologically,IgG4 related cases showed IgG4+ plasma cells/IgG+ plasma cells infiltration and there were more IgG4 related monoclonal antibody expressed when compared to IgG4 unrelated cases.In addition,there were also significant differences in clinical features between the two groups.IgG4 related disease was associated with male gender,higher level of plateletconnt,lymphocytes [(2.4±0.8)×109/L vs (1.4±0.7) ×109/L] count and CRP [(52±60) mg/L vs (15±17) mg/L] levels and lower titer of IgM [(1.2±0.7) g/L vs (1.8±0.8) g/L],antinuclear antibody (56％ vs 87％) and anti-SSB antibodies (13％ vs 54％) (P＜0.05),when compared with IgG4-unrelated cases.There was no significant difference in other indicators (P＞0.05).Conclusion The present study has demonstrated that some of the MD patients misdiagnosed as SS.Some of the laboratory tests such as the level of platelet and lymphocyte count,serum level of CRP,IgM,antinuclear antibody,anti-SSB antibodies,the serum levels of IgG4 and the histopathological presentations in the salivary gland are different between these two disorders.Because of good response to steroid in MD,so laboratory tests and pathological examinations for IgG4 can help to avoid misdiagnosis.

Objective To knock out the entire LuxS gene of Streptococcus mutans UA159 strain via homologous recombination and construct a LuxS-deleted mutant strain of S.mutans.Methods The erythromycin resistance gene(Eymr)was inserted between the two DNA fragments located in the upper and downstream of LuxS gene that had been amplified by PCR.Then the two DNA fragments along with the inserted Eymr were engineered into pUCl9 plasmid to construct the recombination plasmid pUCluxKO.Electrotransformation of S. mutans cells with pUCluxKO-mutant resulted in the isolation of erythromycin resistant S.mutans,transformants,which was then subjected to polymerase chain reaction,Vibrio harveyi BBl70 luminescence bioassay and sequencing analysis.Results Restriction endonuclease analysis showed that pUCluxKOmutant vector had been successfully recombined.The deletion of LuxS of S. mutans mutants was confirmed bv PCR with primers specific for the genes of LuxS and the erythromycin resistance.S.mutans mutant could not induce bioluminescence.indicating the mutant had been successfully recombined.The constructed Chinese S.mutans showed good stability after 20 generations of cultivation.Conclusion The S.mutans gene allelic exchange plasmid is constructed correctively and a LuxS-negative mutant of S.mutans has been constructed.which can be helpful for further study of the role of LuxS in the pathogenesis of S.mutans.

Objective To evaluate the effect of mitochondrial permeability transition pore (mPTP) in lipid emulsion-induced inversion of bupivacaine myocardiotoxicity in rats.Methods H9c2 cells were inoculated in 6-well plates at a density of 105 cells/ml, and were randomly divided into 4 groups (6 wells in each group, 2 ml/well) using a random number table: control group (group C) , bupivacaine group (group B) , lipid emusion + bupivacaine group (group LB) , and lipid emusion + bupivacaine + atractyloside group (group LBA).Phosphate buffer solution 100 μl was added to the culture medium in group C.In group B, bupivacaine was added to the culture medium with the final concentration of 1 mmol/L.In group LB, lipid emusion and bupivacaine were added to the culture medium with the final concentrations of 1％ and 1 mmol/L, respectively.In group LBA, lipid emusion, bupivacaine and atractyloside (an mPTP opener) were added to the culture medium with the final concentrations of 1％, 1 mmol/L and 30 μmol/L, respectively.All the cells were incubated for 24 h.After the end of incubation, the expression of Bcl-2, Bax, phosphorylated Bad (p-Bad) , caspase-3, activated caspase-3, caspase-9,activated caspase-9 and cytochrome c (Cyt c) was detected using Western blot.The expression of Bcl-2 mRNA, Bax mRNA, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was detected using real-time reverse transcriptase polymerase chain reaction.The ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were calculated.Results Compared with group C,the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/ Bcl-2 mRNA were significantly increased, the expression of p-Bad was down-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was up-regulated in group B (P＜0.05) , and no significant change was found in the parameters mentioned above in group LB (P＞0.05).Compared with group B, the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were significantly decreased, the expression of p-Bad was up-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was down-regulated in LB and LBA groups (P＜ 0.05).Compared with group LB, the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were significantly increased, the expression of p-Bad was down-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was up-regulated in group LBA (P ＜ 0.05).Conclusion The mechanism underlying lipid emulsioninduced inversion of bupivacaine myocardiotoxicity is related to inhibited mPTP opening in rats.

Objective To introduce the application of multiplex ligation-dependent probe amplification (MLPA) assays in the diagnosis of patients with hereditary neuropathy with liability to pressure palsies (HNPP).Methods Copy numbers of the exons in peripheral myelin prolein 22 (PMP22) gene,tektin 3 (TEKT3) gene and cytochrome c oxidase assembly protein 10 (COX10) gene were analyzed by MLPA in 8 patients diagnosed with HNPP clinically and 5 normal controls.Results Among the 8 patients,7 patients were identified to have deletion mutations according to their reduced peak area of PMP22 gene,TEKT3 gene and COX10 gene compared with that of normal controls.One patient with normal peak area of PMP22 gene,TEKT3 gene and COX10 gene showed no deletion of these genes.Conclusions MLPA assays can detect the copy numbers of genes in HNPP region through semi-quantitative analysis in a rapid,accurate way,which may be utilized widely in the genetic diagnosis among HNPP patients.