Experimental study on the effect of Kang-Lai-Te induced apoptosis of human hepatoma carcinoma cell HepG2

BACKGROUND: Kang-Lai-Te (KLT) is extracted from the traditional Chinese herbal medicine Semen Coicis, which has been used in China as an effective clinical drug for over a thousand years. It contains numerous ingredients with anti-tumor effects. In our previous studies on transplanted hepatomas in rats, KLT could stop the cells in the G2+M stage of cell cycle and then reduce the number of cells entering the stage G0 and G1, but the mechanism of the anti-proliferative effect was unknown. In this experiment, we examined whether KLT inhibits HepG2 cell growth, if so, tried to explore its mechanism. METHODS: KLT at different concentrations was used for the treatment of hepatocellular carcinoma cells in vitro, respectively. The proliferation inhibitory rate was evaluated by MTT assay, induction of cell apoptosis rate and the protein levels of Fas and Fas ligand (FasL) were determined by flow cytometry (FCM), and the expression of Fas and FasL mRNA was detected by real-time fluorescent quantitative RT-PCR. RESULTS: KLT produced an obvious time and dose-dependent inhibitory effect on HepG2 cells, and marked apoptosis was detected by FCM The protein of Fas increased by 11.01%, 18.71%, 28.71% and 37.15%; the protein of FasL increased by 1.49%, 1.91%, 3.27% and 3.38% in comparison with the control (P<0.05). Real-time fluorescent quantitative RT-PCR showed that treating HepG2 cells with KLT caused the upregulation of Fas and FasL mRNA. CONCLUSION: KLT inhibits HepG2 growth by inducing apoptosis, which may be mediated through activation of the Fas/FasL pathway. (Hepatobiliary Pancreat Dis Int 2009; 8: 267-272)

Expression of NF-kappaB in rotavirus-induced damage to the liver and biliary tract in neonatal mice

BACKGROUND: Biliary atresia, the etiology of which still remains unclear, occurs exclusively in newborns and most are infected with rotavirus. In this study, we aimed to investigate the histopathological patterns of different kinds of rotavirus in the liver and biliary tract of neonatal mice and the expression of NF-kappa B in the liver and biliary tract of infected mice. METHODS: Twenty-three adult mice (8 were male and 15 female) were divided into 8 breeding pairs, and each pair (I male and 2 females) was housed in a cage in a laminar flow hood. Newborn mice, 24-48 hours old were randomly divided into A, B and C groups. The A and B groups were respectively inoculated with MMU18006 and SA11 rotavirus through the intraperitoneal route, while group C as blank control was only inoculated with culture medium. The liver was dissected after 5, 10, 15, 21 and 28 days; the weight of each mouse and the histopathological patterns in the liver were recorded. The expression of NF-kappa B in the liver and intrahepatic bile ducts was detected by immunohistochemical staining and the expression intensity was analyzed with a GT-2 imaging instrument. RESULTS: The average increase in weight of infected mice was significantly slower than that of the normal control, while the growth rate of group A (injected with MMU18006 rotavirus) was slower than that of group B (SA11 rotavirus). In infected mice, the acute and chronic inflammation of liver and intra- and extra-hepatic bile ducts was more significant in group A. Stenosis was found in most intrahepatic bile ducts, and sporadically in extrahepatic bile ducts. The expression of NF-kappa B in infected mice was dramatically higher than that of the normal control, while the expression in group A was higher than in group B. CONCLUSIONS: Significant damage to the liver and biliary tract of neonatal mice can be induced by inoculating MMU18006 rotavirus through the intraperitoneal route, which is very similar to the pathology of biliary atresia in the newborn human. Similar inoculation with SA11 rotavirus can only result in moderate impairment that disappears quickly. The difference of pathogenicity between the two rotaviruses may depend on their differing capacities to increase the expression of NF-kappa B in the liver and biliary tract.

Abdominal pain among children re-evaluation of a diagnostic algorithm

AIM: To re-evaluate the algorithm that has been used forover 40 years for diagnosis of acute abdominal pain amongchildren.METHODS: Among the 937 cases admitted to the surgicalemergency ward in 2000, 656 cases of acute appendicitiswere studied to evaluate the usefulness of the presentalgorithm for its colculated accuracy, false positive and falsenegative rates, the sensitivity and specificity in the instantdiagnosis of various types of acute appendicitis in differentage groups.The algorithm used was established in 1958and revisedror this study in 1999. It includes a 3-step analysisof clinical presentations, i.e.: firstly, a diagnosis of surgicalpain by definite organic abdominal signs; then a diagnosisof the subgroup of surgical condition by special signs; andfinally the diagnosis of the present disease by specific signs.A footnote describes a 'comparative technique″ of abdominalexamination in non-cooperative children.RESULTS: The general accuracy of diagnosis was 92.8 %,overall mortality 0.1% among 973 cases of abdominal painin 2000. 373 attending surgeons and 241 residents includingtrainees joined the diagnosis and treatment with noremarkable difference in the results. The incidence of acuteappendicitis, 656 in 973 cases, was 67.4 % representingthe majority of abdominal pain. In the series of 656 cases,the accuracy of diagnosis of acute appendidtis was 93.6 %,false positive 6.4 %, false negative 0.9 %, sensitivity at firstvisit 82.7 %, specificity for appendicitis 98.0 %, no death ordocumentary complication.CONCLUSION: The present algorithm used for diagnosisof acute abdominal pain is effective and preferable inreducing misdiagnosis and maltreatment at emergency. Theuse of some modern technology should be further explored.

Inverted V-shaped valgus osteotomy for congenital coxa vara

Objective To assess the outcome of inverted V-shaped valgus osteotomy as a treatment for congenital coxa vara in children. Methods Between Feb. 1990 and Jan. 2000,27 patients (35 hips) with congenital coxa vara underwent inverted V-shpaed valgus osteotomy. There were 16 males and 11 females. The average age was 7. 8 years (range:3 to 14 years). They all presented with limp or concomitant pain in the older children. The clinical result was evaluated according to Shi’s criteria. The X-ray films of preoperative, postoperative and the most recent follow-up periods were analyzed. Results Twenty-two patients (28 hips) were followed-up for a mean of 5. 4 years (14 months to 9 years). According to Shi’ s criteria, the excellent and good results accounted for 82. 1 % of cases. The preoperative NS angle, HE angle and ATD value were 94.5° ±9.1° ,67.2° ± 1.8° and - 3.8 mm ± 6. 9 mm respectively whereas those of the immediate postoperative were 148.0° ± 6.5°, 28.6° ± 4.3° and 27.5 mm ± 3. 1 mm; and

Highly selective peripheral neurotomy as a treatment of cerebral palsy spasm

Objective In the patients with spastic cerebral palsy, deformities such as club foot, inversion foot and scissor gait often arise form the imbalance in the adductors muscle group of the thigh and leg and their antagonistic muscle groups. The incerased muscular tension of the former constitutes the rationale of neurotomy. Methods Highly selective peripheral neurotomy in 38 children who suffered from lower limb deformity secondary to unequal nuscular tension. Twenty-one of the children underwent obturator neurotomy, 27 sciatic neurotomy, and 26 tibial neurotomy. Using electric stimulation, the fasciculi involved were decided and severed. The proportion of fasciculi severed depended on the degree of the muscular tension. Results All patients function improved postoperatiely. 80 % of the patients experienced significant improvement. Conclusion Selective peripheral neurotomy is not only more effective and it also has less complication than selective posterior rhizotomy (SPR) in treating cerebral palsy spasm.

Solid cystic papillary tumor of pancreas in children

Objective To estimate the incidence and establish clinicopathological, immunohistochemical , diagnostic and therapeutic features in children with solid cystic papillary tumor (SCPT) of the pancreas in children. Methods Eleven cases (male 1,frmale 10) aging from 9 to 14 years with a mean of 11.1 years with SCPT were analyzed. The chief complains were abdominal pain and palpable mass. The tumors were often located in the head of pancreas. The mean diameter of tumors was 8. 7 cm. The procedures employed included local resections (6), distal pancreatectomy (2) and pancreatico-duodenectomies (3). Routine pathological, histochemical, and immunohistochemical methods were utilized to analyze the specimens. Representative blocks of SCPT specimens were immunostained for NSE, α-1 AT, S - 100, CgA, CK, EMA and insulin. Results The patients were followed-up for 1 to 15 years and were alive postoperatively. Histological examination showed solid with cystic areas and papillary protrusions. Immunohistochemically, the

Early rehabilitative therapy for congenital club foot

To evaluate the results of early rehabilitative therapy for congenital club foot.Methods Eighteen infants (32 feet) with congenital clubfoot were managed with the sequential manipulation,cast and correcting brace in our institute.All of them were followed up.The appearance and function of ankle were recovered.Results The deformities in 9 cases (17 feet) who underwent rehabilitative therapy begining at neonatal period had been totally ameliorated,none of them needed to undergo surgery,and the success rate was up to 100%.On the other hand,the success rate in the other 9 infants (15 feet) who underwent therapy at toddler period was significantly decreased to 60%.And 3 cases had to undergo operations to correct the deformities.Conclusion It is necessary to undergo rehabilitative therapy for infants with congenital clubfoot as early as possible to improve the success rate of treatment.6 refs.

Apoptosis of non-tumor cells contributes to increased serum cytochrome c level in a neuroblastoma xenograft model

Background Neuroblastoma （NB） is one of the most common malignant solid tumors of childhood.It is still not clear whether the apoptosis of tumor cells or the non-tumor cells contributes to the increase of concentration of cytochrome c （Cyt c） in the serum of the cancer patients.The aim of this research was to identify the source of the Cyt c in the serum when the tumor grows up by subcutaneous inoculation of human NB cells into nude mice.Methods We subcutaneously inoculated human NB cells （KP-N-NS） into nude mice and collected the sera of tumor-bearing mice （n=14） and control mice （n=25） 4 weeks later in order to screen for and identify differentially expressed proteins in the serum.Differentially expressed proteins in the serum were screened by surface-enhanced laser desorption/ionization-time-of-fiight （SELDI-TOF） mass spectrometry.Results The relative intensity of a protein having a mass-to-charge ratio （m/z） of 11 609 was 3338.37±3410.85 in the tumor group and 59.84±40.74 in the control group,indicating that the expression level of this protein in the tumor group was 55.8 times higher than that in the control group.Serum proteins were separated and purified by high-performance liquid chromatography （HPLC）.Liquid chromatography coupled with tandem mass spectrometry （LC-MS/MS） was performed to produce peptide mass fingerprints （PMFs）.Spectrum analysis and a database search revealed that the highly expressed protein （m/z=11605.4） from the serum of tumor-bearing mice was the mouse Cyt c.Conclusions Increased concentration of Cyt c in the serum of tumor-bearing nude mice might be partially attributed to the secretion of this protein by non-tumor cells.