Objective:To observe the therapeutic effect and toxicity of local Kanseran injection in combination with chemotherapy in advanced NSCLC patients.Methods:A total of 18 cases was recruited in this study. After desensitization,Kanseran injection was injected into tumor mass through fibrobronchoscope,2 mg every time,once per week. Meanwhile,MVP regimen was administered.Results:Of the 18 patients,1 was completely relieved,7 cases were partially relieved. The response rate was 44.4%. After treatment,when examined by fibrobronchoscope the tumor masses of 15 patients became smaller. The main side effects were fever,alopecia and gastrointestinal reaction,which were mild and tolerable.Conclusion:Local Kanseran injection into tumor mass in conjunction with chemotherapy is effective and safe in the treatment of advanced NSCLC patients with bronchus obstruction.

OBJECTIVETo investigate the mothod and therapeutic efficacy of total hip anthroplasties (THA) for osteoarthritis secondary to Crowe type IV developmental dysplasia of hip in adults.

METHODSFrom May 2006 to December 2013, THA was performed on 15 adult patients (17 hips) with Growe type IV acetabular dysplasia, including 13 females and 2 males, with a mean age of 30.9 years old (22 to 58 years old) and an average preoperative Harris score of (34.0 ± 6.5) points. Traction of the affected limb was not performed before surgery. After extensive release and lengthening of soft tissues, sub-trochanteric osteotomy of the femur was performed, hip joint center was rebuilt and the abduction function was restored.

RESULTSThe patients were followed up with a mean period of 33 months (ranged from 6 months to 5 years). The postoperative Harris score was 85.0 ± 7.3,higher than the preoperative score. The extended length of limb ranged from 1.6 to 5.4 cm, with a mean of (3.42 ± 0.65) cm. The shortening and malformation of the affected limb were corrected in the most patients,with the difference in length of the two legs less than 1.5 cm. After surgery, 1 patient experienced partial sciatic nerve injury, which was largely recovered after 3 months of conservative treatment. One patient experienced complete sciatic nerve injury, which was partially recovered after 6 months of conservative treatment; a foot-drop varus deformity was formed in the distal end of the affected limb, which was improved after tendon transposition and transplantation. Joint pain was relieved, and the joint function was restored significantly. Over the follow-up period, no severe complications such as dislocation, infection, prosthesis loosening, or subsiding occurred.

CONCLUSIONSatisfactory efficacy can be achieved for adult Growe type IV acetabular dysplasia associated with osteoarthritis by THA, with proper soft tissue release and lengthening, sub-trochanteric osteotomy of femur, joint functional restoration, appropriate choice of prosthesis, and careful protection of nerves and vessels.

Adult ; Arthroplasty, Replacement, Hip ; methods ; Female ; Hip Dislocation, Congenital ; complications ; Humans ; Leg Length Inequality ; therapy ; Male ; Middle Aged ; Osteoarthritis, Hip ; surgery

Child ; Humans ; Male ; Muscles ; pathology ; physiopathology ; Myopathies, Structural, Congenital ; diagnosis ; Prognosis

OBJECTIVETo explore the impacts of Yishen Jiangzhuo Granule (YJG) on peripheral blood B-cells and regulatory T-cells (Treg) in patients with chronic renal insufficiency (CRI).

METHODSFifty-three CRI patients were randomly assigned to two groups, the control group and the YJG group. Before and after treatment, the following parameters in blood were detected: the peripheral Treg, percentage (CD19+), activation rate (CD19+ CD69+) and apoptotic rate (AV) of B-lymphocyte by flow cytometry; cytokines (IL-6 and IL-10) by CBA stream protein analyzing system; high sensitivity C-reactive protein (hs-CRP) by scattering turbidimetric analysis; homocysteine (Hcy) by end-point method; hemoglobin (HGB) content by Beckman-Coulter hemo-analyser; blood contents of Ca, phosphate (P), blood urea nitrogen (BUN), creatinine (SCr) and plasma albumin (Alb) by automatic biochemical analyser; and urinary contents of creatinine (UCr) by inverse HPLC. Then the product of calcium-phosphate (Ca x P) was calculated based on blood contents of Ca2 and P and the clearance rate of endogenous creatinine (CCr) was calculated based on blood BUN and SCr.

RESULTSAfter treatment CD19+ and CCr significantly increased (P < 0.01), but AV and SCr decreased in both groups (P < 0.01), with the changes in the YJG group were more significant than those in the control group (P < 0.05); levels of CD19+ CD69+, Treg, IL-6, IL-10, CRP, BUN, P and Ca x P showed no significant change (P > 0.05); levels of Ca2+, HGB and Alb increased as well as of Hcy in both groups (P < 0.05). Correlation analysis: There were negative correlation in CD19+ with AV and Hcy; Alb with AV and Hcy; CCr with CRP, SCr and BUN, while positive correlation existed in SCr with CRP and BUN; and CRP with BUN.

CONCLUSIONSYJG can improve renal function, and delay the progress of renal failure, and it also shows the regulatory effect on B lymphocytes by lowering the apoptosis rate and improving the percentage of CD19+ in patients.

Adult ; Aged ; B-Lymphocytes ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Kidney Failure, Chronic ; drug therapy ; metabolism ; Male ; Middle Aged ; Phytotherapy ; T-Lymphocytes, Regulatory ; metabolism

The increasing prevalence of chronic kidney disease (CKD) is a major global public health concern. Despite the complicated pathogenesis of CKD, renal fibrosis represents the most common pathological condition, comprised of progressive accumulation of extracellular matrix in the diseased kidney. Over the last several decades, tremendous progress in understanding the mechanism of renal fibrosis has been achieved, and corresponding potential therapeutic strategies targeting fibrosis-related signaling pathways are emerging. Importantly, extracellular vesicles (EVs) contribute significantly to renal inflammation and fibrosis by mediating cellular communication. Increasing evidence suggests the potential of EV-based therapy in renal inflammation and fibrosis, which may represent a future direction for CKD therapy.

OBJECTIVETo establish the specific 16S-23S rRNA gene spacer regions pattern in different bacteria using polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), DNA cloning and sequences analysis.

METHODSA pair of primers were selected from highly conserved sequences adjacent to the 16S-23S rRNA spacer region. Bacterial DNA of sixty-one strains of standard bacteria and corresponding clinical isolates representative of 20 genera and 27 species was amplified by PCR, and further studied by RFLP, DNA cloning and sequences analysis. Meanwhile, all specimens were examined by bacterial culturing and PCR-RFLP analysis.

RESULTSThe 27 different standard strains showed one, two, three or more than three bands. The sensitivity of PCR reached 2.5 colony-forming unit (CFU), and there was no cross reaction to the human, fungal or viral genomic DNAs. Fifteen species could be distinguished immediately by PCR, while another 10 species were further identified by Hinf I or Alu I digestion. Klebsiella pneumoniae (Kp) and Enterococcus durans (Ed) could not be differentiated from each other by Alu I or Hinf I digestion. The spacer sequences of the Kp and Ed were 908 bp and 909 bp, respectively, and they differed only at the site of the 779th nucleotide. The former was G, and the latter was A. The 760 - 790 bp sequence of Kp was as follows: CGACTGCACCGCCTCCTAC / GGCCGCGTATTC. The 760 - 790 bp sequence of Ed was as follows: CGACTGCAC CGCCTCCTAC / AGCCGCGTATTC. Only one enzyme XmaIII, could discriminate the two. The cleaving site of XmaIII is C downward arrow GGCCG. Kp DNA was cleaved into 778 bp and 130 bp fragments, while E. durans was not. Of 42 specimens with suspected septicemia, 15 were positive (35.7%) on blood culture, and 27 on PCR (64.29%). The positive rate of PCR was significantly higher than that of blood culture (P < 0.01). Of the six CSF specimens, one was positive for Staphylococcus epidermidis (Se) on culture as well as by PCR, while two specimens which were negative on cultures were positive by PCR and were diagnosed as Se according to its DNA pattern. One specimen was culture-positive for Cryptococcus neoformans (Cn) but was negative by PCR. The other two specimens were negative by both PCR and culture. Fifteen blood samples from healthy children were negative by both blood culture and PCR.

CONCLUSIONSThe method of detecting bacterial 16S-23S rRNA spacer regions using PCR-RFLP techniques was specific, sensitive, rapid and accurate in detecting pathogens in clinical bacterial infections.

Bacterial Infections ; diagnosis ; microbiology ; DNA, Bacterial ; chemistry ; genetics ; DNA, Ribosomal Spacer ; genetics ; Humans ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 16S ; genetics ; RNA, Ribosomal, 23S ; genetics ; Sequence Analysis, DNA

OBJECTIVETo identify 6 major human herpesviruses with consensus primers and to explore its clinical application.

METHODSBased on the highly-homogeneous regions of DNA polymerase gene in human herpesviruses,Two pairs of primer were synthesized. One pair was designed to amplify herpes simplex virus type 1, type 2, Epstein-Barr virus and cytomegalovirus; and another was used to amplify varicella-zoster virus or human herpesvirus 6. Virus species identification was performed by restriction enzyme digestion with BamH I and BstU I. Thirty-eight CSF specimens of clinically diagnosed viral encephalitis,and 49 blood specimens from 27 confirmed cases and 22 clinically diagnosed ones were tested for herpes virus DNA using the PCR-RFLP assay with these primers.

RESULTSThirteen out of 38 CSF specimens (34.2%) were herpes virus positive. All blood specimens from 27 confirmed cases showed positive results, while for 22 clinically diagnosed cases 16 (72.7%) were positive. The types of herpes virus were determined using restriction enzyme digestion with BamH I and BstU I. Two CSF specimens from the patients, who were treated with aciclovir for 2 - 3 days, were still positive for herpes virus DNA by this method. None of the control blood or CSF controls were positive for herpesvirus by PCR.

CONCLUSIONThe PCR-RFLP method used in this study is a specific, sensitive and practicable one for diagnosis of herpes virus infection.

Child ; Cytomegalovirus ; isolation & purification ; Cytomegalovirus Infections ; virology ; DNA Primers ; DNA, Viral ; blood ; cerebrospinal fluid ; Epstein-Barr Virus Infections ; virology ; Female ; Herpesviridae ; isolation & purification ; Herpesviridae Infections ; virology ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Simplexvirus ; isolation & purification

The aim of the present study was to explore the effect of cholecystokinin octapeptide (CCK-8) on [Ca(2+)](i) and its signal transduction mechanism in isolated guinea pig cardiomyocytes. [Ca(2+)](i) was measured by laser scanning confocal microscopy in single ventricular myocytes which were dissociated by enzymatic dissociation method and loaded with Fluo 3-AM. The changes in [Ca(2+)](i) were represented by fluorescent intensity (F(i)) or relative fluorescent intensity (F(i)/F(O)%). The results obtained are as follows. (1) In the normal Tyrode's solution containing 1.0 mmol/ L Ca(2+), CCK-8 (1-10(4) pmol/L) elicited a rapid and marked increase in [Ca(2+)](i). (2) When cardiomyocytes were pretreated with the Ca(2+) chelator EGTA (3 mmol/L) and Ca(2+) channel antagonist nisoldipine (0.5 micromol/L) for 5 min, CCK-8 (10(2)pmol/L) caused a slow and small increase in [Ca(2+)](i) (p< 0.01). (3) Pretreatment with the nonselected CCK- receptor (CCK-R) antagonist proglumide (6 micromol/L) or the tyrosine kinase inhibitor genistein (1 micromol/L) for 5 min could inhibit the increase of [Ca(2+)](i) induced by CCK-8 (10(2) pmol/L) (p<0.01). The results suggest that CCK-8 increases the [Ca(2+)](i) via activating the receptor-operated Ca(2+) channel and eliciting the influx of Ca(2+) in isolated guinea pig cardiomyocytes, in which tyrosine kinase may be involved.
Animals ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Calcium Channels ; drug effects ; Cell Separation ; Guinea Pigs ; Myocytes, Cardiac ; metabolism ; ultrastructure ; Nisoldipine ; pharmacology ; Protein-Tyrosine Kinases ; metabolism ; Signal Transduction ; Sincalide ; pharmacology

The present study was to investigate the effects of agmatine (Agm) on free intracellular calcium concentration ([Ca(2+)]( i )) of isolated rat ventricular myocytes. [Ca(2+)]( i ) was measured by confocal microscopy in single rat ventricular myocytes which were dissociated by enzymatic dissociation method and loaded with Fluo 3-AM. The changes in [Ca(2+)]( i ) were represented by fluorescence intensity (FI) or relative fluorescence intensity (F/F(0)%). The results showed that the control level of FI value of single rat ventricular myocytes was 128.8+/-13.8 and 119.6+/-13.6 in the presence of normal Tyrode's solution containing Ca(2+) 1.0 mmol/L and Ca(2+)-free Tyrode's solution, respectively. There was no difference between these two groups (P>0.05). Agm 0.1, 1, and 10 mmol/L significantly reduced the [Ca(2+)]( i ) in both extracellular solutions in a concentration-dependent manner. The similar effect of Agm on [Ca(2+)]( i ) was also observed in the presence of EGTA 3 mmol/L. KCl 60 mmol/L, PE 30 micromol/L, and Bay-K-8644 10 micromol/L, all these substances induced [Ca(2+)]( i ) elevations in ventricular myocytes. Agm (0.1, 1, and 10 mmol/L) markedly inhibited the increase in [Ca(2+)]( i ) induced by KCl, phenylephrine (PE), and Bay-K-8644. When Ca(2+) waves were produced by increasing extracellular Ca(2+) concentration from 1 to 10 mmol/L, 1 mmol/L Agm could block the propagating waves of elevated [Ca(2+)]( i ), and reduce the velocity and duration of propagating waves. These results suggest that Agm possesses an inhibitory effects on [Ca(2+)]( i ) via blocking voltage-dependent Ca(2+) channel, and possibly by alleviating calcium release from SR in single isolated rat ventricular myocytes.
Agmatine ; pharmacology ; Animals ; Calcium ; metabolism ; Calcium Channels ; drug effects ; Cells, Cultured ; Female ; Heart Ventricles ; cytology ; Male ; Myocytes, Cardiac ; cytology ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley

The effects of low concentration of dihydroouabain (DHO) on intracellular calcium concentration ([Ca(2+)](i)) were investigated in guinea pig ventricular myocytes. [Ca(2+)](i) was detected by confocal microscopy and represented by fluorescent intensity. DHO (1 fmol/L~1 mmol/L) increased [Ca(2+)](i), especially at 10 pmol/L. Nisoldipine, egtazic acid, or tetrodotoxin partially inhibited the effect of 10 pmol/L DHO on [Ca(2+)](i). The effects of DHO remained in the absence of extracellular K(+) and Na(+). These results suggest that low concentration of DHO might increase [Ca(2+)](i) via the receptor-operated Ca(2+) channels, TTX-sensitive Na(+) channels or/and triggering of intracellular calcium release; Na(+)/K(+) pump and Na(+)/Ca(2+) exchange seem not involved in the effect of DHO.
Animals ; Calcium ; metabolism ; Guinea Pigs ; Heart Ventricles ; cytology ; Male ; Myocytes, Cardiac ; cytology ; drug effects ; metabolism ; Ouabain ; analogs & derivatives ; pharmacology ; Patch-Clamp Techniques